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Sample records for brazil virus detection

  1. Serological detection of West Nile virus in horses and chicken from Pantanal, Brazil

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    Melandri, Vanessa; Guimarães, Anthony Érico; Komar, Nicholas; Nogueira, Maurício L; Mondini, Adriano; Fernandez-Sesma, Ana; Alencar, Jeronimo; Bosch, Irene

    2016-01-01

    In an effort to detect West Nile virus (WNV) in Brazil, we sampled serum from horses and chickens from the Pantanal region of the state of Mato Grosso and tested for flavivirus-reactive antibodies by blocking ELISA. The positive samples were further confirmed for serological evidence of WNV infection in three (8%) of the 38 horses and one (3.2%) of the 31 chickens using an 80% plaque-reduction neutralisation test (PRNT80). These results provide evidence of the circulation of WNV in chickens and horses in Pantanal. PMID:23295763

  2. ENTEROPATHOGENS DETECTED IN A DAYCARE CENTER, SOUTHEASTERN BRAZIL: BACTERIA, VIRUS, AND PARASITE RESEARCH

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    Edna Donizetti Rossi Castro

    2015-02-01

    Full Text Available Introduction: The objective of this study was to determine the prevalence and etiological profile of enteropathogens in children from a daycare center. Methods: From October 2010 to February 2011 stool samples from 100 children enrolled in a government daycare center in the municipality of São José do Rio Preto, in the state of São Paulo, were collected and analyzed. Results: A total of 246 bacteria were isolated in 99% of the fecal samples; 129 were in the diarrheal group and 117 in the non-diarrheal group. Seventy-three strains of Escherichia coli were isolated, 19 of Enterobacter, one of Alcaligenes and one of Proteus. There were 14 cases of mixed colonization with Enterobacter and E. coli. Norovirus and Astrovirus were detected in children with clinical signs suggestive of diarrhea. These viruses were detected exclusively among children residing in urban areas. All fecal samples were negative for the presence of the rotavirus species A and C. The presence of Giardia lamblia, Entamoeba coli, Endolimax nana and hookworm was observed. A significant association was found between food consumption outside home and daycare center and the presence of intestinal parasites. Conclusions: For children of this daycare center, intestinal infection due to pathogens does not seem to have contributed to the occurrence of diarrhea or other intestinal symptoms. The observed differences may be due to the wide diversity of geographical, social and economic characteristics and the climate of Brazil, all of which have been reported as critical factors in the modulation of the frequency of different enteropathogens.

  3. Enteropathogens detected in a daycare center, Southeastern Brazil: bacteria, virus, and parasite research.

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    Castro, Edna Donizetti Rossi; Germini, Marcela Cristina Braga Yassaka; Mascarenhas, Joana D'Arc Pereira; Gabbay, Yvone Benchimol; de Lima, Ian Carlos Gomes; Lobo, Patrícia dos Santos; Fraga, Valéria Daltibari; Conceição, Luciana Moran; Machado, Ricardo Luiz Dantas; Rossit, Andréa Regina Baptista

    2015-01-01

    The objective of this study was to determine the prevalence and etiological profile of enteropathogens in children from a daycare center. From October 2010 to February 2011 stool samples from 100 children enrolled in a government daycare center in the municipality of São José do Rio Preto, in the state of São Paulo, were collected and analyzed. A total of 246 bacteria were isolated in 99% of the fecal samples; 129 were in the diarrheal group and 117 in the non-diarrheal group. Seventy-three strains of Escherichia coli were isolated, 19 of Enterobacter, one of Alcaligenes and one of Proteus. There were 14 cases of mixed colonization with Enterobacter and E. coli. Norovirus and Astrovirus were detected in children with clinical signs suggestive of diarrhea. These viruses were detected exclusively among children residing in urban areas. All fecal samples were negative for the presence of the rotavirus species A and C. The presence of Giardia lamblia, Entamoeba coli, Endolimax nana and hookworm was observed. A significant association was found between food consumption outside home and daycare center and the presence of intestinal parasites. For children of this daycare center, intestinal infection due to pathogens does not seem to have contributed to the occurrence of diarrhea or other intestinal symptoms. The observed differences may be due to the wide diversity of geographical, social and economic characteristics and the climate of Brazil, all of which have been reported as critical factors in the modulation of the frequency of different enteropathogens.

  4. Guillain-Barré Syndrome, Acute Disseminated Encephalomyelitis and Encephalitis Associated with Zika Virus Infection in Brazil: Detection of Viral RNA and Isolation of Virus during Late Infection.

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    Brito Ferreira, Maria Lucia; Antunes de Brito, Carlos Alexandre; Moreira, Álvaro José Porto; de Morais Machado, Maria Íris; Henriques-Souza, Adélia; Cordeiro, Marli Tenório; de Azevedo Marques, Ernesto Torres; Pena, Lindomar José

    2017-11-01

    Zika virus (ZIKV) emerged in Brazil in 2015, which was followed by an increase of Guillain-Barre Syndrome (GBS) cases. We report the epidemiological, clinical, and laboratory findings of the first six neurological cases associated with ZIKV in Brazil seen in a reference neurology hospital in Pernambuco, Brazil. In all cases, ZIKV was detected in serum and/or cerebrospinal fluid (CSF) samples. In this case series, four cases were defined as GBS, one as acute disseminated encephalomyelitis (ADEM) and the other as encephalitis. ZIKV was detected in all cases by RT-PCR and virus isolation was successful in two patients. The time between ZIKV acute symptoms and the development of neurological manifestations varied from 3 to 13 days and ZIKV was detected between 15 and 34 days after the initial symptoms. Our results highlight the need to include ZIKV as a differential diagnosis for neurological syndromes in countries with circulation of this arbovirus. Because the viremia in these patients appears to persist longer, direct diagnostic techniques such as RT-PCR and viral isolation should be considered even if it is after the acute phase of viral infection.

  5. Detection of Hepatitis B Virus Antigens in Paraffin-embedded Liver Specimens from the Amazon Region, Brazil

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    Simonetti SRR

    2002-01-01

    Full Text Available Hepatic viscerotomy of paraffin-preserved old specimens, collected in the period from 1934 to 1967, were analyzed by immunohistochemical assays to detect hepatitis B, hepatitis D, dengue and yellow fever virus antigens. The material belongs to the Yellow Fever Collection, Department of Pathology, Instituto Oswaldo Cruz, Rio de Janeiro, Brazil and the cases were diagnosed at that time according to clinical aspects and histopathological findings reporting viral hepatitis, yellow fever, focal necrosis and hepatic atrophy. From the 79 specimens, 69 were collected at the Labrea Region and the other 10 in different other localities in the Amazon Region. The five micra thick histological slices were analyzed for the presence of hepatitis B surface antigen (HBsAg and hepatitis B core antigen (HBcAg by immunoperoxidase technique. An immunofluorescence assay was applied to the detection of hepatitis D, yellow fever and dengue virus antigens. Nine (11.4% histological samples were HBsAg reactive and 5 (6.3% were HBcAg reactive. The oldest reactive sample was from 1934. Viral antigens related to the other pathologies were not detected in this study. Our results confirm that the methodology described may be used to elucidate the aetiology of hepatitis diseases even after a long time of conservation of the specimens.

  6. A human-like H1N2 influenza virus detected during an outbreak of acute respiratory disease in swine in Brazil.

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    Schaefer, Rejane; Rech, Raquel Rubia; Gava, Danielle; Cantão, Mauricio Egídio; da Silva, Marcia Cristina; Silveira, Simone; Zanella, Janice Reis Ciacci

    2015-01-01

    Passive monitoring for detection of influenza A viruses (IAVs) in pigs has been carried out in Brazil since 2009, detecting mostly the A(H1N1)pdm09 influenza virus. Since then, outbreaks of acute respiratory disease suggestive of influenza A virus infection have been observed frequently in Brazilian pig herds. During a 2010-2011 influenza monitoring, a novel H1N2 influenza virus was detected in nursery pigs showing respiratory signs. The pathologic changes were cranioventral acute necrotizing bronchiolitis to subacute proliferative and purulent bronchointerstitial pneumonia. Lung tissue samples were positive for both influenza A virus and A(H1N1)pdm09 influenza virus based on RT-qPCR of the matrix gene. Two IAVs were isolated in SPF chicken eggs. HI analysis of both swine H1N2 influenza viruses showed reactivity to the H1δ cluster. DNA sequencing was performed for all eight viral gene segments of two virus isolates. According to the phylogenetic analysis, the HA and NA genes clustered with influenza viruses of the human lineage (H1-δ cluster, N2), whereas the six internal gene segments clustered with the A(H1N1)pdm09 group. This is the first report of a reassortant human-like H1N2 influenza virus derived from pandemic H1N1 virus causing an outbreak of respiratory disease in pigs in Brazil. The emergence of a reassortant IAV demands the close monitoring of pigs through the full-genome sequencing of virus isolates in order to enhance genetic information about IAVs circulating in pigs.

  7. Persistent Zika Virus Detection in Semen in a Traveler Returning to the United Kingdom from Brazil, 2016.

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    Gaskell, Katherine M; Houlihan, Catherine; Nastouli, Eleni; Checkley, Anna M

    2017-01-01

    Zika virus is normally transmitted by mosquitos, but cases of sexual transmission have been reported. We describe a patient with symptomatic Zika virus infection in whom the virus was detected in semen for 92 days. Our findings support recommendations for 6 months of barrier contraceptive use after symptomatic Zika virus infection.

  8. Vector Surveillance for Dengue Virus Detection in the Archipelago of Fernando de Noronha, Brazil.

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    Barbosa, P P; Guedes, D R D; Melo-Santos, M A V; Cordeiro, M T; Acioli, R V; Batista, C A V; Gonçalves, L S M; Souza, M F M; Araújo, Y V; Magalhães, F J R; Regis, L; Ayres, C F J

    2016-05-01

    Aedes aegypti (L.) has become an efficient vector of important arboviruses due to its anthropophilic and domiciliary behaviors. Since the 1980s, dengue affects thousands of people every year in Brazil; in Fernando de Noronha (FN), a touristic archipelago, dengue cases have occurred since 2001. Once Ae. aegypti populations are well established in the inhabited areas of FN, the threat of dengue or another arbovirus epidemic is continuously imminent. This study aimed to monitor the DENV serotypes in mosquito samples collected in FN, where at least one resident was clinically diagnosed as dengue patient. Entomological surveillance was conducted in 2011 and 2012. Mosquitoes were sorted by sex and location and were stored in pools. DENV detection was performed using polymerase chain reaction with reverse transcription (RT-PCR) and the Platelia Dengue NS1 Ag. RNA integrity was checked by RT-PCR using rpL8 primers, and the minimum infection rate (MIR) was calculated. In total, 339 pools were analyzed, and only one was positive (DENV-1) by Multiplex RT-PCR (MIR = 1.53). When considering only pools with RNA integrity, the MIR was 2.92. Using the Platelia kit, the MIR was 9.18 (considering all the pools) and 17.54 (only 140 pools with RNA integrity). Our results showed the importance of a constant entomological surveillance in that area, the need to improve storage and transportation protocols, and an endogenous control in the RT-PCR to avoid false-negative results. Finally, our study indicated that the NS1-Ag detection was the most sensitive method and should be used routinely for DENV surveillance in mosquitoes if the serotype identification is not required. © The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  9. First detection of natural infection of Aedes aegypti with Zika virus in Brazil and throughout South America

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    Anielly Ferreira-de-Brito

    Full Text Available Zika virus (ZIKV has caused a major epidemic in Brazil and several other American countries. ZIKV is an arbovirus whose natural vectors during epidemics have been poorly determined. In this study, 1,683 mosquitoes collected in the vicinity of ZIKV suspected cases in Rio de Janeiro, Brazil, from June 2015 to May 2016 were screened for natural infection by using molecular methods. Three pools of Aedes aegypti were found with the ZIKV genome, one of which had only one male. This finding supports the occurrence of vertical and/or venereal transmission of ZIKV in Ae. aegypti in nature. None of the examined Ae. albopictus and Culex quinquefasciatus was positive. This is the first report of natural infection by ZIKV in mosquitoes in Brazil and other South American countries. So far, Ae. aegypti is the only confirmed vector of ZIKV during the ongoing Pan-American epidemics.

  10. DETECTION OF BOVINE RESPIRATORY SYNCYTIAL VIRUS IN CALVES OF RIO GRANDE DO SUL, BRAZIL

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    Ivan Paulo Demartini Gonçalves

    1993-12-01

    Full Text Available During 20 months of the 1987-1990 period, lung tissue samples from 351 calves were obtained at a slaughterhouse. These calves were from counties nearby Porto Alegre. The direct and indirect fluorescent antibody tests (FAT using polyclonal and monoclonal antibody conjugates were performed on frozen lung sections. Eighteen (5.13% of the calf lung samples were positive for the Bovine Respiratory Syncytial Virus (BRSV. The BRSV was isolated from FAT positive samples.

  11. Hepatitis B virus genotype E detected in Brazil in an African patient who is a frequent traveler

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    R Sitnik

    2007-12-01

    Full Text Available Genotype E of hepatitis B virus (HBV has not been described in Brazil and is found mainly in Africa. Genotype A is the most prevalent in Brazil, and genotypes B, C, D, and F have already been reported. We report here an HBV genotype E-infected patient and some characterization of surface (S protein, DNA polymerase (P and precore/core (preC/C coding regions based on the viral genome. The patient is a 31-year-old black man with chronic hepatitis B who was born and raised in Angola. He has been followed by a hepatologist in São Paulo, Brazil, since November 2003, and he is a frequent traveler to Latin America, Africa, and Europe. In 2003, he was diagnosed with HBV infection and started treatment with lamivudine with the later addition of adefovir dipivoxil. No known risk factor was identified. Serologically, he is HBsAg and anti-HBe positive, but HBeAg and anti-HBs negative. DNA sequence analysis of the S/P region confirmed that this patient is infected with genotype E, subtype ayw4. The preC/C region showed G1896A and G1899A mutations but no mutations in the basal core promoter. Nucleotide substitutions common in genotype E were also observed (C1772, T1858 and A1757. Although this is not an autochthonous case and there is no evidence of further spread, the description of this case in Brazil highlights the current risk of viral genotypes spreading with unprecedented speed due to constant travel around the world.

  12. Detection and partial molecular characterization of Grapevine fleck virus, Grapevine virus D, Grapevine leafroll-associated virus -5 and -6 infecting grapevines in Brazil.

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    Fajardo,Thor Vinícius Martins; Eiras,Marcelo; Nickel,Osmar; Dubiela,Carla Rosa; Souto,Eliezer Rodrigues de

    2012-01-01

    Grapevine fleck, rugose wood and leafroll are three grapevine viral diseases whose causal agents (or associated viruses) respectively are Grapevine fleck virus (GFkV), Grapevine virus D (GVD) and Grapevine leafroll-associated virus 5 and 6 (GLRaV-5 and -6). The objective of this work was to perform a partial molecular characterization of local isolates of these four viral species that infect grapevines. The nucleotide and deduced amino acid sequences of complete genes of the coat protein (CP)...

  13. SEROLOGICAL DETECTION OF HEPATITIS A VIRUS IN FREE-RANGING NEOTROPICAL PRIMATES (Sapajus spp., Alouatta caraya) FROM THE PARANÁ RIVER BASIN, BRAZIL.

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    Svoboda, Walfrido Kühl; Soares, Manoel do Carmo Pereira; Alves, Max Moreira; Rocha, Tatiana Carneiro; Gomes, Eliane Carneiro; Menoncin, Fabiana; Batista, Paulo Mira; Silva, Lineu Roberto da; Headley, Selwyn Arlington; Hilst, Carmen Lúcia Scortecci; Aguiar, Lucas M; Ludwig, Gabriela; Passos, Fernando de Camargo; Souza, Júlio Cesar de; Navarro, Italmar Teodorico

    2016-01-01

    Nonhuman primates are considered as the natural hosts of Hepatitis A virus (HAV), as well as other pathogens, and can serve as natural sentinels to investigate epizootics and endemic diseases that are of public health importance. During this study, blood samples were collected from 112 Neotropical primates (NTPs) (Sapajus nigritus and S. cay, n = 75; Alouatta caraya, n = 37) trap-captured at the Paraná River basin, Brazil, located between the States of Paraná and Mato Grosso do Sul. Anti-HAV IgG antibodies were detected in 4.5% (5/112) of NTPs, specifically in 6.7% (5/75) of Sapajus spp. and 0% (0/37) of A. caraya. In addition, all samples were negative for the presence of IgM anti-HAV antibodies. These results suggest that free-ranging NTPs were exposed to HAV within the geographical regions evaluated.

  14. Dengue Virus Type 4, Manaus, Brazil

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    Naveca, Felipe Gomes; de Souza Bastos, Michele; do Nascimento Melo, Miriam; de Souza Viana, Suziane; Mourão, Maria Paula Gomes; Costa, Cristóvão Alves; Farias, Izeni Pires

    2008-01-01

    We report dengue virus type 4 (DENV-4) in Amazonas, Brazil. This virus was isolated from serum samples of 3 patients treated at a tropical medicine reference center in Manaus. All 3 cases were confirmed by serologic and molecular tests; 1 patient was co-infected with DENV-3 and DENV-4. PMID:18394292

  15. First report of autochthonous transmission of Zika virus in Brazil

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    Camila Zanluca

    2015-06-01

    Full Text Available In the early 2015, several cases of patients presenting symptoms of mild fever, rash, conjunctivitis and arthralgia were reported in the northeastern Brazil. Although all patients lived in a dengue endemic area, molecular and serological diagnosis for dengue resulted negative. Chikungunya virus infection was also discarded. Subsequently, Zika virus (ZIKV was detected by reverse transcription-polymerase chain reaction from the sera of eight patients and the result was confirmed by DNA sequencing. Phylogenetic analysis suggests that the ZIKV identified belongs to the Asian clade. This is the first report of ZIKV infection in Brazil.

  16. Vaccine protection against Zika virus from Brazil.

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    Larocca, Rafael A; Abbink, Peter; Peron, Jean Pierre S; Zanotto, Paolo M de A; Iampietro, M Justin; Badamchi-Zadeh, Alexander; Boyd, Michael; Ng'ang'a, David; Kirilova, Marinela; Nityanandam, Ramya; Mercado, Noe B; Li, Zhenfeng; Moseley, Edward T; Bricault, Christine A; Borducchi, Erica N; Giglio, Patricia B; Jetton, David; Neubauer, George; Nkolola, Joseph P; Maxfield, Lori F; De La Barrera, Rafael A; Jarman, Richard G; Eckels, Kenneth H; Michael, Nelson L; Thomas, Stephen J; Barouch, Dan H

    2016-08-25

    Zika virus (ZIKV) is a flavivirus that is responsible for the current epidemic in Brazil and the Americas. ZIKV has been causally associated with fetal microcephaly, intrauterine growth restriction, and other birth defects in both humans and mice. The rapid development of a safe and effective ZIKV vaccine is a global health priority, but very little is currently known about ZIKV immunology and mechanisms of immune protection. Here we show that a single immunization with a plasmid DNA vaccine or a purified inactivated virus vaccine provides complete protection in susceptible mice against challenge with a strain of ZIKV involved in the outbreak in northeast Brazil. This ZIKV strain has recently been shown to cross the placenta and to induce fetal microcephaly and other congenital malformations in mice. We produced DNA vaccines expressing ZIKV pre-membrane and envelope (prM-Env), as well as a series of deletion mutants. The prM-Env DNA vaccine, but not the deletion mutants, afforded complete protection against ZIKV, as measured by absence of detectable viraemia following challenge, and protective efficacy correlated with Env-specific antibody titers. Adoptive transfer of purified IgG from vaccinated mice conferred passive protection, and depletion of CD4 and CD8 T lymphocytes in vaccinated mice did not abrogate this protection. These data demonstrate that protection against ZIKV challenge can be achieved by single-shot subunit and inactivated virus vaccines in mice and that Env-specific antibody titers represent key immunologic correlates of protection. Our findings suggest that the development of a ZIKV vaccine for humans is likely to be achievable.

  17. Detection of the B"-GWGR variant in the southernmost region of Brazil: unveiling the complexity of the human immunodeficiency virus-1 subtype B epidemic

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    Dennis Maletich Junqueira

    2013-09-01

    Full Text Available Typical human immunodeficiency virus-1 subtype B (HIV-1B sequences present a GPGR signature at the tip of the variable region 3 (V3 loop; however, unusual motifs harbouring a GWGR signature have also been isolated. Although epidemiological studies have detected this variant in approximately 17-50% of the total infections in Brazil, the prevalence of B"-GWGR in the southernmost region of Brazil is not yet clear. This study aimed to investigate the C2-V3 molecular diversity of the HIV-1B epidemic in southernmost Brazil. HIV-1 seropositive patients were ana-lysed at two distinct time points in the state of Rio Grande do Sul (RS98 and RS08 and at one time point in the state of Santa Catarina (SC08. Phylogenetic analysis classified 46 individuals in the RS98 group as HIV-1B and their molecular signatures were as follows: 26% B"-GWGR, 54% B-GPGR and 20% other motifs. In the RS08 group, HIV-1B was present in 32 samples: 22% B"-GWGR, 59% B-GPGR and 19% other motifs. In the SC08 group, 32 HIV-1B samples were found: 28% B"-GWGR, 59% B-GPGR and 13% other motifs. No association could be established between the HIV-1B V3 signatures and exposure categories in the HIV-1B epidemic in RS. However, B-GPGR seemed to be related to heterosexual individuals in the SC08 group. Our results suggest that the established B"-GWGR epidemics in both cities have similar patterns, which is likely due to their geographical proximity and cultural relationship.

  18. Detection of the B"-GWGR variant in the southernmost region of Brazil: unveiling the complexity of the human immunodeficiency virus-1 subtype B epidemic.

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    Junqueira, Dennis Maletich; Medeiros, Rúbia Marília de; Leite, Thaysse Cristina Neiva Ferreira; Guimarães, Monick Lindenmeyer; Gräf, Tiago; Pinto, Aguinaldo Roberto; Almeida, Sabrina Esteves de Matos

    2013-09-01

    Typical human immunodeficiency virus-1 subtype B (HIV-1B) sequences present a GPGR signature at the tip of the variable region 3 (V3) loop; however, unusual motifs harbouring a GWGR signature have also been isolated. Although epidemiological studies have detected this variant in approximately 17-50% of the total infections in Brazil, the prevalence of B"-GWGR in the southernmost region of Brazil is not yet clear. This study aimed to investigate the C2-V3 molecular diversity of the HIV-1B epidemic in southernmost Brazil. HIV-1 seropositive patients were ana-lysed at two distinct time points in the state of Rio Grande do Sul (RS98 and RS08) and at one time point in the state of Santa Catarina (SC08). Phylogenetic analysis classified 46 individuals in the RS98 group as HIV-1B and their molecular signatures were as follows: 26% B"-GWGR, 54% B-GPGR and 20% other motifs. In the RS08 group, HIV-1B was present in 32 samples: 22% B"-GWGR, 59% B-GPGR and 19% other motifs. In the SC08 group, 32 HIV-1B samples were found: 28% B"-GWGR, 59% B-GPGR and 13% other motifs. No association could be established between the HIV-1B V3 signatures and exposure categories in the HIV-1B epidemic in RS. However, B-GPGR seemed to be related to heterosexual individuals in the SC08 group. Our results suggest that the established B"-GWGR epidemics in both cities have similar patterns, which is likely due to their geographical proximity and cultural relationship.

  19. Detection and molecular characterisation of a diagnosis escape variant associated with occult hepatitis B virus in Brazil

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    Ricardo Wagner de Almeida

    Full Text Available BACKGROUND Many studies have identified mutations in the hepatitis B surface antigen (HBsAg as important factors limiting the ability of commercial serological assays to detect this viral antigen. However, an association between mutations in the HBsAg gene and the occurrence of occult HBV infection (OBI in patients has not been established. OBJECTIVES To detect hepatitis B virus (HBV DNA in patients with anti-HBc as a unique serological marker, a previously published, cost-effective TaqMan-based real-time polymerase chain reaction (PCR test with minor groove binding probes was adapted for use in this study. The current study also aimed to investigate HBsAg mutations and genotypes of HBV in OBI at the Viral Hepatitis Ambulatory Clinic in Rio de Janeiro to determine any possible association. METHODS Intra-assay and inter-assay reproducibility were determined, and the mean coefficient of variation values obtained were 2.07 and 3.5, respectively. Probit analysis indicated that the 95% detection level was 25 IU/mL. The prevalence of OBI was investigated in 35 serum samples with an ‘anti-HBc alone’ profile from individuals who attended our clinic between 2011 and 2013. FINDINGS HBV DNA was detected in only one sample, resulting in an OBI rate of 2.9%. Nucleotide sequencing of the pre-S/S region was performed to genotype and analyse mutations within the HBsAg gene of this HBV DNA. The HBV in the OBI case was classified as sub-genotype A1, and a sequence analysis of the small S gene revealed 12 mutations in the major hydrophilic region compared to the consensus A1 sequence. Most of these mutations occurred in amino acid residues that have been reported as clinically relevant because they have been implicated in vaccine escape and/or inability to detect HBsAg by commercial serological assays. MAIN CONCLUSIONS Our study suggests the importance of specific HBsAg mutations, different from those in D, B, and C genotypes, in sub-genotype A1 HBV associated

  20. PRESENCE OF RESPIRATORY VIRUSES IN EQUINES IN BRAZIL

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    Dalva Assunção Portari Mancini

    2014-06-01

    Full Text Available Equines are susceptible to respiratory viruses such as influenza and parainfluenza. Respiratory diseases have adversely impacted economies all over the world. This study was intended to determine the presence of influenza and parainfluenza viruses in unvaccinated horses from some regions of the state of São Paulo, Brazil. Blood serum collected from 72 equines of different towns in this state was tested by hemagglutination inhibition test to detect antibodies for both viruses using the corresponding antigens. About 98.6% (71 and 97.2% (70 of the equines responded with antibody protective titers (≥ 80 HIU/25µL H7N7 and H3N8 subtypes of influenza A viruses, respectively. All horses (72 also responded with protective titers (≥ 80 HIU/25µL against the parainfluenza virus. The difference between mean antibody titers to H7N7 and H3N8 subtypes of influenza A viruses was not statistically significant (p > 0.05. The mean titers for influenza and parainfluenza viruses, on the other hand, showed a statistically significant difference (p < 0.001. These results indicate a better antibody response from equines to parainfluenza 3 virus than to the equine influenza viruses. No statistically significant differences in the responses against H7N7 and H3N8 subtypes of influenza A and parainfluenza 3 viruses were observed according to the gender (female, male or the age (≤ 2 to 20 years-old groups. This study provides evidence of the concomitant presence of two subtypes of the equine influenza A (H7N7 and H3N8 viruses and the parainfluenza 3 virus in equines in Brazil. Thus, it is advisable to vaccinate equines against these respiratory viruses.

  1. Identification of circo-like virus-Brazil genomic sequences in raw sewage from the metropolitan area of São Paulo: evidence of circulation two and three years after the first detection.

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    Castrignano, Silvana Beres; Nagasse-Sugahara, Teresa Keico; Garrafa, Patrícia; Monezi, Telma Alves; Barrella, Karina Medici; Mehnert, Dolores Ursula

    2017-03-01

    Two novel viruses named circo-like virus-Brazil (CLV-BR) hs1 and hs2 were previously discovered in a Brazilian human fecal sample through metagenomics. CLV-BR hs1 and hs2 possess a small circular DNA genome encoding a replication initiator protein (Rep), and the two genomes exhibit 92% nucleotide identity with each other. Phylogenetic analysis based on the Rep protein showed that CLV-BRs do not cluster with circoviruses, nanoviruses, geminiviruses or cycloviruses. The aim of this study was to search for CLV-BR genomes in sewage and reclaimed water samples from the metropolitan area of São Paulo, Brazil, to verify whether the first detection of these viruses was an isolated finding. Sewage and reclaimed water samples collected concomitantly during the years 2005-2006 were purified and concentrated using methodologies designed for the study of viruses. A total of 177 treated reclaimed water samples were grouped into five pools, as were 177 treated raw sewage samples. Nucleic acid extraction, polymerase chain reaction (PCR) amplification and Sanger sequencing were then performed.e. genomes were detected in two pools of sewage samples, p6 and p9. Approximately 28% and 51% of the CLV-BR genome was amplified from p6 and p9, respectively, including 76% of the Rep gene. The detected genomes are most likely related to CLV-BR hs1. Comparative analysis showed several synonymous substitutions within Rep-encoding sequences, suggesting purifying selection for this gene, as has been observed for other eukaryotic circular Rep-encoding single-stranded DNA (CRESS-DNA) viruses. The results therefore indicated that CLV-BR has continued to circulate in Brazil two and three years after first being detected.

  2. Identification of circo-like virus-Brazil genomic sequences in raw sewage from the metropolitan area of São Paulo: evidence of circulation two and three years after the first detection

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    Silvana Beres Castrignano

    Full Text Available BACKGROUND Two novel viruses named circo-like virus-Brazil (CLV-BR hs1 and hs2 were previously discovered in a Brazilian human fecal sample through metagenomics. CLV-BR hs1 and hs2 possess a small circular DNA genome encoding a replication initiator protein (Rep, and the two genomes exhibit 92% nucleotide identity with each other. Phylogenetic analysis based on the Rep protein showed that CLV-BRs do not cluster with circoviruses, nanoviruses, geminiviruses or cycloviruses. OBJECTIVE The aim of this study was to search for CLV-BR genomes in sewage and reclaimed water samples from the metropolitan area of São Paulo, Brazil, to verify whether the first detection of these viruses was an isolated finding. METHODS Sewage and reclaimed water samples collected concomitantly during the years 2005-2006 were purified and concentrated using methodologies designed for the study of viruses. A total of 177 treated reclaimed water samples were grouped into five pools, as were 177 treated raw sewage samples. Nucleic acid extraction, polymerase chain reaction (PCR amplification and Sanger sequencing were then performed.e FINDINGS CLV-BR genomes were detected in two pools of sewage samples, p6 and p9. Approximately 28% and 51% of the CLV-BR genome was amplified from p6 and p9, respectively, including 76% of the Rep gene. The detected genomes are most likely related to CLV-BR hs1. Comparative analysis showed several synonymous substitutions within Rep-encoding sequences, suggesting purifying selection for this gene, as has been observed for other eukaryotic circular Rep-encoding single-stranded DNA (CRESS-DNA viruses. MAIN CONCLUSION The results therefore indicated that CLV-BR has continued to circulate in Brazil two and three years after first being detected.

  3. Detection of the first incidence of Akodon paranaensis naturally infected with the Jabora virus strain (Hantavirus in Brazil

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    Renata Carvalho de Oliveira

    2012-05-01

    Full Text Available We characterised hantaviruses circulating in different Akodon rodent species collected in midwestern Santa Catarina (SC, southern Brazil, where the Jabora hantavirus (JABV strain was first identified in Akodon montensis. Genetic and phylogenetic analyses based on a partial S segment indicated that, in SC, Akodon paranaensis and A. montensis carried the same type of hantavirus. Additionally, we conducted the first genomic characterisation of the complete S segment from the Brazilian JABV strain. This is the first report of A. paranaensis infected with the JABV.

  4. Zika virus RNA detection in asymptomatic blood donors during an outbreak in the northeast region of São Paulo State, Brazil, 2016.

    Science.gov (United States)

    Slavov, Svetoslav N; Hespanhol, Marta R; Rodrigues, Evandra S; Levi, José E; Ubiali, Eugênia M A; Covas, Dimas T; Kashima, Simone

    2017-12-01

    In 2015, there was a large Zika virus (ZIKV) outbreak in Brazil. The proportion of asymptomatic infections is very high, and it is possible for transfusion-transmitted ZIKV (TT-ZIKV) infection to occur. The prevalence of asymptomatic ZIKV infection among Brazilian blood donors during this epidemic outbreak is unknown. Plasma samples obtained between October 2015 and May 2016 from 1393 volunteer blood donors were tested for ZIKV RNA. The viral load was quantified using an in-house standard curve. Additionally, positive ZIKV RNA samples were tested for anti-ZIKV immunoglobulin (Ig)M and anti-ZIKV IgG. Of the 1393 blood samples, ZIKV RNA was detected in 37 (n = 37/1393; 2.7%). The median infection viral load detected was 7714 copies/mL (ranging from 135-124,220 copies/mL). The majority of the positive samples (70.3%) exhibited a viral load of approximately 103 copies/mL. Six samples that were positive for ZIKV RNA were also positive for anti-ZIKV IgM and IgG (n = 6/37; 13.5%). This is the first study evaluating the prevalence of ZIKV RNA among Brazilian blood donors, which was relatively high and might lead to TT-ZIKV infection. It is unclear whether the simultaneous presence of anti-ZIKV IgM and IgG in RNA-positive donations or the viral load influences transfusion transmission of the infection. This study also adds to the global understanding of ZIKV prevalence in blood donors during outbreaks and the transfusion impact of the infection. © 2017 AABB.

  5. Dengue virus detection by using reverse transcription-polymerase chain reaction in saliva and progeny of experimentally infected Aedes albopictus from Brazil

    Directory of Open Access Journals (Sweden)

    Márcia Gonçalves de Castro

    2004-12-01

    Full Text Available Oral susceptibility and vertical transmission of dengue virus type 2 (DENV-2 in an Aedes albopictus sample from Rio de Janeiro was estimated. The infection (36.7% and transmission (83.3% rates for Ae. albopictus were higher than those of an Ae. aegypti colony used as control, 32.8 and 60%, respectively. Fourth instar larvae and females descendants of 48.5 and 39.1% of experimentally infected Ae. albopictus showed to harbor the virus. The oral susceptibility and the high capacity to assure vertical transmission exhibited by Ae. albopictus from Brazil reinforce that this species may play a role in the maintenance of the virus in nature and be a threat for dengue control in the country.

  6. Dengue virus detection by using reverse transcription-polymerase chain reaction in saliva and progeny of experimentally infected Aedes albopictus from Brazil.

    Science.gov (United States)

    Castro, Márcia Gonçalves de; Nogueira, Rita Maria Ribeiro; Schatzmayr, Hermann Gonçalves; Miagostovich, Marize Pereira; Lourenço-de-Oliveira, Ricardo

    2004-12-01

    Oral susceptibility and vertical transmission of dengue virus type 2 (DENV-2) in an Aedes albopictus sample from Rio de Janeiro was estimated. The infection (36.7%) and transmission (83.3%) rates for Ae. albopictus were higher than those of an Ae. aegypti colony used as control, 32.8 and 60%, respectively. Fourth instar larvae and females descendants of 48.5 and 39.1% of experimentally infected Ae. albopictus showed to harbor the virus. The oral susceptibility and the high capacity to assure vertical transmission exhibited by Ae. albopictus from Brazil reinforce that this species may play a role in the maintenance of the virus in nature and be a threat for dengue control in the country.

  7. Computer Viruses: Pathology and Detection.

    Science.gov (United States)

    Maxwell, John R.; Lamon, William E.

    1992-01-01

    Explains how computer viruses were originally created, how a computer can become infected by a virus, how viruses operate, symptoms that indicate a computer is infected, how to detect and remove viruses, and how to prevent a reinfection. A sidebar lists eight antivirus resources. (four references) (LRW)

  8. Emergence and potential for spread of Chikungunya virus in Brazil.

    Science.gov (United States)

    Nunes, Marcio Roberto Teixeira; Faria, Nuno Rodrigues; de Vasconcelos, Janaina Mota; Golding, Nick; Kraemer, Moritz U G; de Oliveira, Layanna Freitas; Azevedo, Raimunda do Socorro da Silva; da Silva, Daisy Elaine Andrade; da Silva, Eliana Vieira Pinto; da Silva, Sandro Patroca; Carvalho, Valéria Lima; Coelho, Giovanini Evelim; Cruz, Ana Cecília Ribeiro; Rodrigues, Sueli Guerreiro; Vianez, Joao Lídio da Silva Gonçalves; Nunes, Bruno Tardelli Diniz; Cardoso, Jedson Ferreira; Tesh, Robert B; Hay, Simon I; Pybus, Oliver G; Vasconcelos, Pedro Fernando da Costa

    2015-04-30

    In December 2013, an outbreak of Chikungunya virus (CHIKV) caused by the Asian genotype was notified in the Caribbean. The outbreak has since spread to 38 regions in the Americas. By September 2014, the first autochthonous CHIKV infections were confirmed in Oiapoque, North Brazil, and in Feira de Santana, Northeast Brazil. We compiled epidemiological and clinical data on suspected CHIKV cases in Brazil and polymerase-chain-reaction-based diagnostic was conducted on 68 serum samples from patients with symptom onset between April and September 2014. Two imported and four autochthonous cases were selected for virus propagation, RNA isolation, full-length genome sequencing, and phylogenetic analysis. We then followed CDC/PAHO guidelines to estimate the risk of establishment of CHIKV in Brazilian municipalities. We detected 41 CHIKV importations and 27 autochthonous cases in Brazil. Epidemiological and phylogenetic analyses indicated local transmission of the Asian CHIKV genotype in Oiapoque. Unexpectedly, we also discovered that the ECSA genotype is circulating in Feira de Santana. The presumed index case of the ECSA genotype was an individual who had recently returned from Angola and developed symptoms in Feira de Santana. We estimate that, if CHIKV becomes established in Brazil, transmission could occur in 94% of municipalities in the country and provide maps of the risk of importation of each strain of CHIKV in Brazil. The etiological strains associated with the early-phase CHIKV outbreaks in Brazil belong to the Asian and ECSA genotypes. Continued surveillance and vector mitigation strategies are needed to reduce the future public health impact of CHIKV in the Americas.

  9. Novel Viruses Isolated from Mosquitoes in Pantanal, Brazil.

    Science.gov (United States)

    Pauvolid-Corrêa, Alex; Solberg, Owen; Couto-Lima, Dinair; Nogueira, Rita Maria; Langevin, Stanley; Komar, Nicholas

    2016-11-03

    Genomic sequences are described from five novel viruses and divergent strains of Brejeira and Guaico Culex viruses from mosquitoes collected in Pantanal, Brazil, in 2010. Copyright © 2016 Pauvolid-Corrêa et al.

  10. Establishment and cryptic transmission of Zika virus in Brazil and the Americas

    Science.gov (United States)

    Faria, N. R.; Quick, J.; Claro, I. M.; Thézé, J.; de Jesus, J. G.; Giovanetti, M.; Kraemer, M. U. G.; Hill, S. C.; Black, A.; da Costa, A. C.; Franco, L. C.; Silva, S. P.; Wu, C.-H.; Raghwani, J.; Cauchemez, S.; Du Plessis, L.; Verotti, M. P.; de Oliveira, W. K.; Carmo, E. H.; Coelho, G. E.; Santelli, A. C. F. S.; Vinhal, L. C.; Henriques, C. M.; Simpson, J. T.; Loose, M.; Andersen, K. G.; Grubaugh, N. D.; Somasekar, S.; Chiu, C. Y.; Muñoz-Medina, J. E.; Gonzalez-Bonilla, C. R.; Arias, C. F.; Lewis-Ximenez, L. L.; Baylis, S. A.; Chieppe, A. O.; Aguiar, S. F.; Fernandes, C. A.; Lemos, P. S.; Nascimento, B. L. S.; Monteiro, H. A. O.; Siqueira, I. C.; de Queiroz, M. G.; de Souza, T. R.; Bezerra, J. F.; Lemos, M. R.; Pereira, G. F.; Loudal, D.; Moura, L. C.; Dhalia, R.; França, R. F.; Magalhães, T.; Marques, E. T.; Jaenisch, T.; Wallau, G. L.; de Lima, M. C.; Nascimento, V.; de Cerqueira, E. M.; de Lima, M. M.; Mascarenhas, D. L.; Neto, J. P. Moura; Levin, A. S.; Tozetto-Mendoza, T. R.; Fonseca, S. N.; Mendes-Correa, M. C.; Milagres, F. P.; Segurado, A.; Holmes, E. C.; Rambaut, A.; Bedford, T.; Nunes, M. R. T.; Sabino, E. C.; Alcantara, L. C. J.; Loman, N. J.; Pybus, O. G.

    2017-06-01

    Transmission of Zika virus (ZIKV) in the Americas was first confirmed in May 2015 in northeast Brazil. Brazil has had the highest number of reported ZIKV cases worldwide (more than 200,000 by 24 December 2016) and the most cases associated with microcephaly and other birth defects (2,366 confirmed by 31 December 2016). Since the initial detection of ZIKV in Brazil, more than 45 countries in the Americas have reported local ZIKV transmission, with 24 of these reporting severe ZIKV-associated disease. However, the origin and epidemic history of ZIKV in Brazil and the Americas remain poorly understood, despite the value of this information for interpreting observed trends in reported microcephaly. Here we address this issue by generating 54 complete or partial ZIKV genomes, mostly from Brazil, and reporting data generated by a mobile genomics laboratory that travelled across northeast Brazil in 2016. One sequence represents the earliest confirmed ZIKV infection in Brazil. Analyses of viral genomes with ecological and epidemiological data yield an estimate that ZIKV was present in northeast Brazil by February 2014 and is likely to have disseminated from there, nationally and internationally, before the first detection of ZIKV in the Americas. Estimated dates for the international spread of ZIKV from Brazil indicate the duration of pre-detection cryptic transmission in recipient regions. The role of northeast Brazil in the establishment of ZIKV in the Americas is further supported by geographic analysis of ZIKV transmission potential and by estimates of the basic reproduction number of the virus.

  11. Establishment and cryptic transmission of Zika virus in Brazil and the Americas.

    Science.gov (United States)

    Faria, N R; Quick, J; Claro, I M; Thézé, J; de Jesus, J G; Giovanetti, M; Kraemer, M U G; Hill, S C; Black, A; da Costa, A C; Franco, L C; Silva, S P; Wu, C-H; Raghwani, J; Cauchemez, S; du Plessis, L; Verotti, M P; de Oliveira, W K; Carmo, E H; Coelho, G E; Santelli, A C F S; Vinhal, L C; Henriques, C M; Simpson, J T; Loose, M; Andersen, K G; Grubaugh, N D; Somasekar, S; Chiu, C Y; Muñoz-Medina, J E; Gonzalez-Bonilla, C R; Arias, C F; Lewis-Ximenez, L L; Baylis, S A; Chieppe, A O; Aguiar, S F; Fernandes, C A; Lemos, P S; Nascimento, B L S; Monteiro, H A O; Siqueira, I C; de Queiroz, M G; de Souza, T R; Bezerra, J F; Lemos, M R; Pereira, G F; Loudal, D; Moura, L C; Dhalia, R; França, R F; Magalhães, T; Marques, E T; Jaenisch, T; Wallau, G L; de Lima, M C; Nascimento, V; de Cerqueira, E M; de Lima, M M; Mascarenhas, D L; Neto, J P Moura; Levin, A S; Tozetto-Mendoza, T R; Fonseca, S N; Mendes-Correa, M C; Milagres, F P; Segurado, A; Holmes, E C; Rambaut, A; Bedford, T; Nunes, M R T; Sabino, E C; Alcantara, L C J; Loman, N J; Pybus, O G

    2017-06-15

    Transmission of Zika virus (ZIKV) in the Americas was first confirmed in May 2015 in northeast Brazil. Brazil has had the highest number of reported ZIKV cases worldwide (more than 200,000 by 24 December 2016) and the most cases associated with microcephaly and other birth defects (2,366 confirmed by 31 December 2016). Since the initial detection of ZIKV in Brazil, more than 45 countries in the Americas have reported local ZIKV transmission, with 24 of these reporting severe ZIKV-associated disease. However, the origin and epidemic history of ZIKV in Brazil and the Americas remain poorly understood, despite the value of this information for interpreting observed trends in reported microcephaly. Here we address this issue by generating 54 complete or partial ZIKV genomes, mostly from Brazil, and reporting data generated by a mobile genomics laboratory that travelled across northeast Brazil in 2016. One sequence represents the earliest confirmed ZIKV infection in Brazil. Analyses of viral genomes with ecological and epidemiological data yield an estimate that ZIKV was present in northeast Brazil by February 2014 and is likely to have disseminated from there, nationally and internationally, before the first detection of ZIKV in the Americas. Estimated dates for the international spread of ZIKV from Brazil indicate the duration of pre-detection cryptic transmission in recipient regions. The role of northeast Brazil in the establishment of ZIKV in the Americas is further supported by geographic analysis of ZIKV transmission potential and by estimates of the basic reproduction number of the virus.

  12. Zika virus infections imported from Brazil to Portugal, 2015

    Directory of Open Access Journals (Sweden)

    L. Zé-Zé

    2016-01-01

    Here, we present the clinical and laboratory aspects related to the first four imported human cases of Zika virus in Portugal from Brazil, and alert, regarding the high level of traveling between Portugal and Brazil, and the ongoing expansion of this virus in the Americas, for the threat for Zika virus introduction in Europe and the possible introduction to Madeira Island where Aedes aegypti is present.

  13. Detection and identification of dengue virus isolates from Brazil by a simplified reverse transcription - polymerase chain reaction (RT-PCR method

    Directory of Open Access Journals (Sweden)

    FIGUEIREDO Luiz Tadeu Moraes

    1997-01-01

    Full Text Available We show here a simplified RT-PCR for identification of dengue virus types 1 and 2. Five dengue virus strains, isolated from Brazilian patients, and yellow fever vaccine 17DD as a negative control, were used in this study. C6/36 cells were infected and supernatants were collected after 7 days. The RT-PCR, done in a single reaction vessel, was carried out following a 1/10 dilution of virus in distilled water or in a detergent mixture containing Nonidet P40. The 50 µl assay reaction mixture included 50 pmol of specific primers amplifying a 482 base pair sequence for dengue type 1 and 210 base pair sequence for dengue type 2. In other assays, we used dengue virus consensus primers having maximum sequence similarity to the four serotypes, amplifying a 511 base pair sequence. The reaction mixture also contained 0.1 mM of the four deoxynucleoside triphosphates, 7.5 U of reverse transcriptase, 1U of thermostable Taq DNA polymerase. The mixture was incubated for 5 minutes at 37ºC for reverse transcription followed by 30 cycles of two-step PCR amplification (92ºC for 60 seconds, 53ºC for 60 seconds with slow temperature increment. The PCR products were subjected to 1.7% agarose gel electrophoresis and visualized by UV light after staining with ethidium bromide solution. Low virus titer around 10 3, 6 TCID50/ml was detected by RT-PCR for dengue type 1. Specific DNA amplification was observed with all the Brazilian dengue strains by using dengue virus consensus primers. As compared to other RT-PCRs, this assay is less laborious, done in a shorter time, and has reduced risk of contamination

  14. Zika virus displacement by a chikungunya outbreak in Recife, Brazil.

    Directory of Open Access Journals (Sweden)

    Tereza Magalhaes

    2017-11-01

    Full Text Available Several arboviruses, including dengue virus (DENV, Zika virus (ZIKV and chikungunya virus (CHIKV, transmitted by Aedes mosquitoes, circulate in northeast Brazil. Diseases caused by these viruses are of great public health relevance, however, their epidemiological features in areas where the three viruses co-circulate are scarce. Here, we present analyses of molecular and serological diagnostics in a prospective study of acute febrile patients recruited from May 2015 to May 2016 in Recife, Brazil.Two hundred sixty-three acute febrile patients with symptoms suggestive of an arboviral disease who attended an urgent heath care clinic in the Recife Metropolitan Region in northeast Brazil were enrolled. Acute and convalescent blood samples were collected and tested using molecular and serological assays for infection with DENV, ZIKV and CHIKV.Quantitative real-time reverse-transcriptase polymerase chain reactions (qRTPCR performed on acute phase sera detected no patients positive for DENV, but 26 (9.9% positive for ZIKV and 132 (50.2% positive for CHIKV. There were a few suspected and only one confirmed dengue case. Specific serological assays for ZIKV and CHIKV confirmed the qRTPCR data. Analyses of DENV IgM and IgG ELISAs in the context of qRTPCR results suggested high levels of cross reactive antibodies in ZIKV-positive samples. Results from neutralization assays highly corroborated those from qRTPCR and ZIKV ELISA, indicating very few positive DENV cases. ZIKV infections were temporally clustered in the first months of the study and started to decrease concomitantly with an increase in CHIKV infections in August 2015. The proportion of CHIKV infections increased significantly in September 2015 and remained high until the end of the study period, with an average of 84.7% of recruited patients being diagnosed from August 2015 to May 2016. ZIKV infections exhibited a female bias and the cases were spread over the study site, while CHIKV cases had a

  15. Zika virus displacement by a chikungunya outbreak in Recife, Brazil.

    Science.gov (United States)

    Magalhaes, Tereza; Braga, Cynthia; Cordeiro, Marli T; Oliveira, Andre L S; Castanha, Priscila M S; Maciel, Ana Paula R; Amancio, Nathalia M L; Gouveia, Pollyanne N; Peixoto-da-Silva, Valter J; Peixoto, Thaciana F L; Britto, Helena; Lima, Priscilla V; Lima, Andreza R S; Rosenberger, Kerstin D; Jaenisch, Thomas; Marques, Ernesto T A

    2017-11-01

    Several arboviruses, including dengue virus (DENV), Zika virus (ZIKV) and chikungunya virus (CHIKV), transmitted by Aedes mosquitoes, circulate in northeast Brazil. Diseases caused by these viruses are of great public health relevance, however, their epidemiological features in areas where the three viruses co-circulate are scarce. Here, we present analyses of molecular and serological diagnostics in a prospective study of acute febrile patients recruited from May 2015 to May 2016 in Recife, Brazil. Two hundred sixty-three acute febrile patients with symptoms suggestive of an arboviral disease who attended an urgent heath care clinic in the Recife Metropolitan Region in northeast Brazil were enrolled. Acute and convalescent blood samples were collected and tested using molecular and serological assays for infection with DENV, ZIKV and CHIKV. Quantitative real-time reverse-transcriptase polymerase chain reactions (qRTPCR) performed on acute phase sera detected no patients positive for DENV, but 26 (9.9%) positive for ZIKV and 132 (50.2%) positive for CHIKV. There were a few suspected and only one confirmed dengue case. Specific serological assays for ZIKV and CHIKV confirmed the qRTPCR data. Analyses of DENV IgM and IgG ELISAs in the context of qRTPCR results suggested high levels of cross reactive antibodies in ZIKV-positive samples. Results from neutralization assays highly corroborated those from qRTPCR and ZIKV ELISA, indicating very few positive DENV cases. ZIKV infections were temporally clustered in the first months of the study and started to decrease concomitantly with an increase in CHIKV infections in August 2015. The proportion of CHIKV infections increased significantly in September 2015 and remained high until the end of the study period, with an average of 84.7% of recruited patients being diagnosed from August 2015 to May 2016. ZIKV infections exhibited a female bias and the cases were spread over the study site, while CHIKV cases had a male bias and

  16. Validation of Plant Virus Detection

    NARCIS (Netherlands)

    Schadewijk, van A.R.; Meekes, E.T.M.; Verbeek, M.; Verhoeven, J.Th.J.

    2011-01-01

    Validation of test methods is required for laboratories seeking ISO 17025 accreditation. Recently developed manuals help choosing relevant performance characteristics to be studied for qualitative tests common in plant virus detection. For routine testing in certification schemes additional

  17. Detection of rotavirus in dogs with diarrhea in Brazil

    Directory of Open Access Journals (Sweden)

    Gabbay Yvone B.

    2003-01-01

    Full Text Available Rotavirus was detected by the enzyme-linked immunosorbent assay (ELISA in the faeces of a diarrheic dog. Virus particles with morphology typical of rotavirus were visualized by direct electron microscopy. This sample was subsequently tested for the four main human serotypes (G1-G4, by ELISA with monoclonal antibodies. G genotyping was attempted by RT-PCR using G1-G6 and G8-G11 primers but no positive results could be yielded. Also using RT-PCR it was possible to characterize this canine strain as belonging to P[ 3] genotype. This is the first canine rotavirus detected in Brazil.

  18. Chikungunya virus infection: report of the first case diagnosed in Rio de Janeiro, Brazil

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    Isabella Gomes Cavalcanti de Albuquerque

    2012-02-01

    Full Text Available Initially diagnosed in Africa and Asia, the Chikungunya virus has been detected in the last three years in the Caribbean, Italy, France, and the United States of America. Herein, we report the first case for Rio de Janeiro, Brazil, in 2010.

  19. Detection of hepatitis C virus in patients with terminal renal disease undergoing dialysis in southern Brazil: prevalence, risk factors, genotypes, and viral load dynamics in hemodialysis patients.

    Science.gov (United States)

    Vidales-Braz, Beatris Maria; da Silva, Naylê Maria Oliveira; Lobato, Rubens; Germano, Fabiana Nunes; da Mota, Luiza Dias; Barros, Elvino J G; de Martinez, Ana Maria Barral

    2015-02-03

    Hepatitis C (HCV) is a serious public health issue, and it is estimated that 3% of the world's population is infected. Patients in hemodialysis units have an increased risk for contracting HCV, and high prevalence rates have been found in hemodialysis units around the world. This study is aimed at determining the prevalence of HCV in patients with terminal chronic renal disease (tCRD) who have been submitted to hemodialysis and peritoneal dialysis in southern Brazil to characterize the most prevalent genotypes, the viral load, and possible risk factors and to assess the validity between the ELISA and RT-PCR detection methods. Of 320 patients from three dialysis units, 318 participated in this study. According to the medical records, 55 patients were reactive to HCV, as determined via ELISA. All 318 samples were submitted to RT-PCR and genotyped using an Abbott Realtime m2000 system. Data obtained through a questionnaire and chemical variables were associated with the HCV. The prevalence of HCV was 18.24% (58), and the concordance between the HCV serology and the RT-PCR was 94%. Three patients were diagnosed to be negative for HCV using the ELISA assay but positive when using RT-PCR. Genotype 1 was the most prevalent (46.7%) genotype, within which subtype 1a was the most frequent (74.1%). One of the risk factors associated with HCV infection was the length of time that the patient had been undergoing hemodialysis treatments (p hemodialysis (p hemodialysis indicates a possible destruction or gripping of viral particles to the dialyzer membrane.

  20. First report of autochthonous transmission of Zika virus in Brazil

    National Research Council Canada - National Science Library

    Zanluca, Camila; Melo, Vanessa Campos Andrade de; Mosimann, Ana Luiza Pamplona; Santos, Glauco Igor Viana Dos; Santos, Claudia Nunes Duarte Dos; Luz, Kleber

    2015-01-01

    .... Chikungunya virus infection was also discarded. Subsequently, Zika virus (ZIKV) was detected by reverse transcription-polymerase chain reaction from the sera of eight patients and the result was confirmed by DNA sequencing...

  1. Rabies virus in Molossus molossus (Chiroptera: Molossidae in the State of Pernambuco, Northeastern Brazil

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    Luiz Augustinho Menezes da Silva

    2011-08-01

    Full Text Available Rabies virus was detected in bats (Molossus molossus from an urban area in the City of Recife, State of Pernambuco, Brazil. Four individuals were found during the day in visible, non-habitual places, lying on the ground, but still alive. No contact occurred with people or animals. Of these, only two were identified; it was not possible to identify two specimens, since they were incinerated prior to identification. Diagnosis was positive by direct immunofluorescence and intracerebral inoculation in mice. This study presents the first instance in which the virus was detected in insectivorous bats in the State of Pernambuco.

  2. Assessment of enteric viruses in a sewage treatment plant located in Porto Alegre, southern Brazil

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    AD. Vecchia

    Full Text Available In order to verify the microbial quality of the influents and effluents of one STP from southern Brazil, an eight-month survey was conducted to examine the presence of total and fecal coliforms and of adenovirus (HAdV, enterovirus (EV, genogroup A rotaviruses (GARV and Torque teno virus (TTV, in treated effluent samples from São João/Navegantes STP, Porto Alegre (Brazil. A total of 16 samples were collected, eight of influent (raw sewage, prior to treatment, and the other eight of the effluent (post-treatment sewage. Total and fecal coliform levels ranging from 3.6 × 10(4 to 4.4 × 10(7 MPN/100 mL and 2.9 × 10³ to 1.7 × 10(7 MPN/100 mL, were detected in all samples. In raw sewage, HAdV (25% and GARV (28.6% viral genomes were detected. The analysis of effluent samples revealed the presence of HAdV (50%, EV (37.5%, and TTV (12.5% genomic fragments. All samples, regardless of the month analysed, presented detection of a least one virus genus, except for in April. Higher virus detection rates were observed in treated sewage samples (62.5%, and in 80% of them (effluent positive samples HAdV was detected. Results showed that improvements in sewage monitoring and treatment processes are necessary to reduce the viral and bacterial load on the environment in southern Brazil. To the knowledge of the authors, this is the first study showing the monitoring of viral genomes in influent and effluent samples from a STP located in Porto Alegre (Rio Grande do Sul, Brazil, southern Brazil.

  3. Prevalence of Torque teno virus in healthy donors of Paraná State, southern Brazil.

    Science.gov (United States)

    Mazzola, Jocimara Costa; Saito, Patrícia Keiko; Yamakawa, Roger Haruki; Watanabe, Maria Angélica Ehara; da Silva Junior, Waldir Veríssimo; Matta, Alessandra Cristina Gobbi; Borelli, Sueli Donizete

    2015-01-01

    To determine the prevalence of the Torque teno virus in healthy donors in the northern and northwestern regions of the state of Paraná, southern Brazil. The Torque teno virus was detected by a nested polymerase chain reaction using a set of oligoprimers for the N22 region. The prevalence of the virus was 69% in 551 healthy blood donors in southern Brazil. There was no statistically significant difference between the presence of the virus and the variables gender, ethnicity and marital status. There was significant difference in the prevalence of the virus regarding the age of the donors (p-value=0.024) with a higher incidence (74.7%) in 18- to 24-year-old donors. A high prevalence of Torque teno virus was observed in the population studied. Further studies are needed to elucidate the routes of contamination and the clinical implications of the virus in the healthy population. Copyright © 2015 Associação Brasileira de Hematologia, Hemoterapia e Terapia Celular. Published by Elsevier Editora Ltda. All rights reserved.

  4. Prevalence of Torque teno virus in healthy donors of Paraná State, southern Brazil

    Directory of Open Access Journals (Sweden)

    Jocimara Costa Mazzola

    2015-10-01

    Full Text Available OBJECTIVE: To determine the prevalence of the Torque teno virus in healthy donors in the northern and northwestern regions of the state of Paraná, southern Brazil.METHODS: The Torque teno virus was detected by a nested polymerase chain reaction using a set of oligoprimers for the N22 region.RESULTS: The prevalence of the virus was 69% in 551 healthy blood donors in southern Brazil. There was no statistically significant difference between the presence of the virus and the variables gender, ethnicity and marital status. There was significant difference in the prevalence of the virus regarding the age of the donors (p-value = 0.024 with a higher incidence (74.7% in 18- to 24-year-old donors.CONCLUSION: A high prevalence of Torque teno virus was observed in the population studied. Further studies are needed to elucidate the routes of contamination and the clinical implications of the virus in the healthy population.

  5. Influenza A Viruses of Human Origin in Swine, Brazil

    Science.gov (United States)

    Schaefer, Rejane; Gava, Danielle; Cantão, Maurício Egídio; Ciacci-Zanella, Janice Reis

    2015-01-01

    The evolutionary origins of the influenza A(H1N1)pdm09 virus that caused the first outbreak of the 2009 pandemic in Mexico remain unclear, highlighting the lack of swine surveillance in Latin American countries. Although Brazil has one of the largest swine populations in the world, influenza was not thought to be endemic in Brazil’s swine until the major outbreaks of influenza A(H1N1)pdm09 in 2009. Through phylogenetic analysis of whole-genome sequences of influenza viruses of the H1N1, H1N2, and H3N2 subtypes collected in swine in Brazil during 2009–2012, we identified multiple previously uncharacterized influenza viruses of human seasonal H1N2 and H3N2 virus origin that have circulated undetected in swine for more than a decade. Viral diversity has further increased in Brazil through reassortment between co-circulating viruses, including A(H1N1)pdm09. The circulation of multiple divergent hemagglutinin lineages challenges the design of effective cross-protective vaccines and highlights the need for additional surveillance. PMID:26196759

  6. Dengue virus detection by using reverse transcription-polymerase chain reaction in saliva and progeny of experimentally infected Aedes albopictus from Brazil

    OpenAIRE

    Márcia Gonçalves Castro; Rita Maria Ribeiro Nogueira; Hermann Gonçalves Schatzmayr; Marize Pereira Miagostovich; Ricardo Lourenço-de-Oliveira

    2004-01-01

    Oral susceptibility and vertical transmission of dengue virus type 2 (DENV-2) in an Aedes albopictus sample from Rio de Janeiro was estimated. The infection (36.7%) and transmission (83.3%) rates for Ae. albopictus were higher than those of an Ae. aegypti colony used as control, 32.8 and 60%, respectively. Fourth instar larvae and females descendants of 48.5 and 39.1% of experimentally infected Ae. albopictus showed to harbor the virus. The oral susceptibility and the high capacity to assure ...

  7. Possible Association Between Zika Virus Infection and Microcephaly - Brazil, 2015.

    Science.gov (United States)

    Schuler-Faccini, Lavinia; Ribeiro, Erlane M; Feitosa, Ian M L; Horovitz, Dafne D G; Cavalcanti, Denise P; Pessoa, André; Doriqui, Maria Juliana R; Neri, Joao Ivanildo; Neto, Joao Monteiro de Pina; Wanderley, Hector Y C; Cernach, Mirlene; El-Husny, Antonette S; Pone, Marcos V S; Serao, Cassio L C; Sanseverino, Maria Teresa V

    2016-01-29

    In early 2015, an outbreak of Zika virus, a flavivirus transmitted by Aedes mosquitoes, was identified in northeast Brazil, an area where dengue virus was also circulating. By September, reports of an increase in the number of infants born with microcephaly in Zika virus-affected areas began to emerge, and Zika virus RNA was identified in the amniotic fluid of two women whose fetuses had been found to have microcephaly by prenatal ultrasound. The Brazil Ministry of Health (MoH) established a task force to investigate the possible association of microcephaly with Zika virus infection during pregnancy and a registry for incident microcephaly cases (head circumference ≥2 standard deviations [SD] below the mean for sex and gestational age at birth) and pregnancy outcomes among women suspected to have had Zika virus infection during pregnancy. Among a cohort of 35 infants with microcephaly born during August-October 2015 in eight of Brazil's 26 states and reported to the registry, the mothers of all 35 had lived in or visited Zika virus-affected areas during pregnancy, 25 (71%) infants had severe microcephaly (head circumference >3 SD below the mean for sex and gestational age), 17 (49%) had at least one neurologic abnormality, and among 27 infants who had neuroimaging studies, all had abnormalities. Tests for other congenital infections were negative. All infants had a lumbar puncture as part of the evaluation and cerebrospinal fluid (CSF) samples were sent to a reference laboratory in Brazil for Zika virus testing; results are not yet available. Further studies are needed to confirm the association of microcephaly with Zika virus infection during pregnancy and to understand any other adverse pregnancy outcomes associated with Zika virus infection. Pregnant women in Zika virus-affected areas should protect themselves from mosquito bites by using air conditioning, screens, or nets when indoors, wearing long sleeves and pants, using permethrin-treated clothing and gear

  8. Zika Virus: Transmission, Detection, Control, and Prevention

    Science.gov (United States)

    Sharma, Anshika; Lal, Sunil K.

    2017-01-01

    Zika virus (ZIKV) is a mosquito-borne Flavivirus discovered in Uganda in the 1940s. To date, three major ZIKV outbreaks have been reported. ZIKV infections have known to be primarily asymptomatic while causing mild illness in a few cases. However, the recent emergence and spread of ZIKV in the Americas has resulted in the declaration of “Public Health Emergency of International Concern” due to the potential association between the infection and prenatal microcephaly or other brain anomalies. In Brazil, a 20-fold increase in prenatal microcephaly cases and 19% increase in Guillain-Barré Syndrome (GBS) cases were reported in 2015, as compared to the preceding year. The probable deleterious effects of ZIKV infection prompt the urgent development of diagnostics and therapeutics. To this end, the existing evidences supporting the increasingly common prenatal microcephaly and GBS association and the current known ZIKV transmission dynamics, modes of detection (molecular and serology-based), and current control strategies are summarized in this review. This review also emphasizes the importance of understanding ZIKV transmission in order to design a sensitive yet cost and time-efficient detection technique. Development of an efficient detection technique would subsequently allow for better surveillance and control of ZIKV infection. Currently, limited literature is available on the pathogenesis of ZIKV, hence, focusing on the modes of ZIKV transmission could potentially contribute to the understanding of the disease spectrum and formulation of targeted treatment and control. PMID:28217114

  9. Nhumirim virus, a novel flavivirus isolated from mosquitoes from the Pantanal, Brazil

    Science.gov (United States)

    Solberg, Owen; Couto-Lima, Dinair; Kenney, Joan; Serra-Freire, Nicolau; Brault, Aaron; Nogueira, Rita; Langevin, Stanley; Komar, Nicholas

    2016-01-01

    We describe the isolation of a novel flavivirus, isolated from a pool of mosquitoes identified as Culex (Culex) chidesteri collected in 2010 in the Pantanal region of west-central Brazil. The virus is herein designated Nhumirim virus (NHUV) after the name of the ranch from which the mosquito pool was collected. Flavivirus RNA was detected by real-time RT-PCR of homogenized mosquitoes and from the corresponding C6/36 culture supernatant. Based on full-genome sequencing, the virus isolate was genetically distinct from but most closely related to Barkedji virus (BJV), a newly described flavivirus from Senegal. Phylogenetic analysis demonstrated that NHUV grouped with mosquito-borne flaviviruses forming a clade with BJV. This clade may be genetically intermediate between the Culex-borne flaviviruses amplified by birds and the insect-only flaviviruses. PMID:25252815

  10. Early Evidence for Zika Virus Circulation among Aedes aegypti Mosquitoes, Rio de Janeiro, Brazil

    Science.gov (United States)

    Ayllón, Tania; Campos, Renata de Mendonça; Brasil, Patrícia; Morone, Fernanda Cristina; Câmara, Daniel Cardoso Portela; Meira, Guilherme Louzada Silva; Tannich, Egbert; Yamamoto, Kristie Aimi; Carvalho, Marilia Sá; Pedro, Renata Saraiva; Cadar, Daniel; Ferreira, Davis Fernandes; Honório, Nildimar Alves

    2017-01-01

    During 2014–2016, we conducted mosquito-based Zika virus surveillance in Rio de Janeiro, Brazil. Results suggest that Zika virus was probably introduced into the area during May–November 2013 via multiple in-country sources. Furthermore, our results strengthen the hypothesis that Zika virus in the Americas originated in Brazil during October 2012–May 2013. PMID:28628464

  11. Serologic survey of West Nile virus in horses from Central-West, Northeast and Southeast Brazil

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    Jaqueline Raymondi Silva

    2013-11-01

    Full Text Available Since the emergence of West Nile virus (WNV in North America in 1999, there have been several reports of WNV activity in Central and South American countries. To detect WNV in Brazil, we performed a serological survey of horses from different regions of Brazil using recombinant peptides from domain III of WNV. Positive samples were validated with the neutralisation test. Our results showed that of 79 ELISA-positive horses, nine expressed WNV-specific neutralising antibodies. Eight of the infected horses were from the state of Mato Grosso do Sul and one was from the state of Paraíba. Our results provide additional evidence for the emergence of WNV in Brazil and for its circulation in multiple regions of the country.

  12. Zika virus infection in a traveller returning to Europe from Brazil, March 2015.

    Science.gov (United States)

    Zammarchi, L; Tappe, D; Fortuna, C; Remoli, M E; Günther, S; Venturi, G; Bartoloni, A; Schmidt-Chanasit, J

    2015-06-11

    We report a case of laboratory-confirmed Zika virus infection imported into Europe from the Americas. The patient developed fever, rash, and oedema of hands and feet after returning to Italy from Brazil in late March 2015. The case highlights that, together with chikungunya virus and dengue virus, three major arboviruses are now co-circulating in Brazil. These arboviruses represent a burden for the healthcare systems in Brazil and other countries where competent mosquito vectors are present.

  13. High frequency of hepatitis E virus infection in swine from South Brazil and close similarity to human HEV isolates

    Directory of Open Access Journals (Sweden)

    Ana Maria Passos-Castilho

    Full Text Available Abstract Hepatitis E virus is responsible for acute and chronic liver infections worldwide. Swine hepatitis E virus has been isolated in Brazil, and a probable zoonotic transmission has been described, although data are still scarce. The aim of this study was to investigate the frequency of hepatitis E virus infection in pigs from a small-scale farm in the rural area of Paraná State, South Brazil. Fecal samples were collected from 170 pigs and screened for hepatitis E virus RNA using a duplex real-time RT-PCR targeting a highly conserved 70 nt long sequence within overlapping parts of ORF2 and ORF3 as well as a 113 nt sequence of ORF2. Positive samples with high viral loads were subjected to direct sequencing and phylogenetic analysis. hepatitis E virus RNA was detected in 34 (20.0% of the 170 pigs following positive results in at least one set of screening real-time RT-PCR primers and probes. The swine hepatitis E virus strains clustered with the genotype hepatitis E virus-3b reference sequences in the phylogenetic analysis and showed close similarity to human hepatitis E virus isolates previously reported in Brazil.

  14. Circulation of different lineages of Dengue virus 2, genotype American/Asian in Brazil: dynamics and molecular and phylogenetic characterization.

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    Betânia Paiva Drumond

    Full Text Available The American/Asian genotype of Dengue virus type 2 (DENV-2 was introduced into the Americas in the 80's. Although there is no data showing when this genotype was first introduced into Brazil, it was first detected in Brazil in 1990. After which the virus spread throughout the country and major epidemics occurred in 1998, 2007/08 and 2010. In this study we sequenced 12 DENV-2 genomes obtained from serum samples of patients with dengue fever residing in São José do Rio Preto, São Paulo (SJRP/SP, Brazil, in 2008. The whole open reading frame or envelope sequences were used to perform phylogenetic, phylogeographic and evolutionary analyses. Isolates from SJRP/SP were grouped within one lineage (BR3 close to isolates from Rio de Janeiro, Brazil. Isolates from SJRP were probably introduced there at least in 2007, prior to its detection in the 2008 outbreak. DENV-2 circulation in Brazil is characterized by the introduction, displacement and circulation of three well-defined lineages in different times, most probably from the Caribbean. Thirty-seven unique amino acid substitutions were observed among the lineages, including seven amino acid differences in domains I to III of the envelope protein. Moreover, we dated here, for the first time, the introduction of American/Asian genotype into Brazil (lineage BR1 to 1988/89, followed by the introduction of lineages BR2 (1998-2000 and BR3 (2003-05. Our results show a delay between the introduction and detection of DENV-2 lineages in Brazil, reinforcing the importance and need for surveillance programs to detect and trace the evolution of these viruses. Additionally, Brazilian DENV-2 differed in genetic diversity, date of introduction and geographic origin and distribution in Brazil, and these are important factors for the evolution, dynamics and control of dengue.

  15. Concurrent detection of other respiratory viruses in children shedding viable human respiratory syncytial virus.

    Science.gov (United States)

    Gagliardi, T B; Paula, F E; Iwamoto, M A; Proença-Modena, J L; Santos, A E; Camara, A A; Cervi, M C; Cintra, O A L; Arruda, E

    2013-10-01

    Human respiratory syncytial virus (HRSV) is an important cause of respiratory disease. The majority of studies addressing the importance of virus co-infections to the HRSV-disease have been based on the detection of HRSV by RT-PCR, which may not distinguish current replication from prolonged shedding of remnant RNA from previous HRSV infections. To assess whether co-detections of other common respiratory viruses are associated with increased severity of HRSV illnesses from patients who were shedding viable-HRSV, nasopharyngeal aspirates from children younger than 5 years who sought medical care for respiratory infections in Ribeirão Preto (Brazil) were tested for HRSV by immunofluorescence, RT-PCR and virus isolation in cell culture. All samples with viable-HRSV were tested further by PCR for other respiratory viruses. HRSV-disease severity was assessed by a clinical score scale. A total of 266 samples from 247 children were collected and 111 (42%) were HRSV-positive. HRSV was isolated from 70 (63%), and 52 (74%) of them were positive for at least one additional virus. HRSV-positive diseases were more severe than HRSV-negative ones, but there was no difference in disease severity between patients with viable-HRSV and those HRSV-positives by RT-PCR. Co-detection of other viruses did not correlate with increased disease severity. HRSV isolation in cell culture does not seem to be superior to RT-PCR to distinguish infections associated with HRSV replication in studies of clinical impact of HRSV. A high rate of co-detection of other respiratory viruses was found in samples with viable-HRSV, but this was not associated with more severe HRSV infection. Copyright © 2013 Wiley Periodicals, Inc.

  16. Hepatitis E virus in liver and bile samples from slaughtered pigs of Brazil

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    Noemi Rovaris Gardinali

    2012-11-01

    Full Text Available The objective of this study was to detect and identify hepatitis E virus (HEV strains in liver and bile samples from slaughtered pigs in the state of Paraná, Brazil. Liver and bile samples were collected from 118 asymptomatic adult pigs at a slaughterhouse in a major Brazilian pork production area. The samples were assayed using a nested reverse transcription-polymerase chain reaction protocol with primer sets targeting open reading frames (ORF1 and 2 of the HEV genome. HEV RNA was detected in two (1.7% liver samples and one (0.84% bile sample using both primers sets. The HEV strains were classified as genotype 3b on the basis of their nucleotide sequences. These data suggest that healthy pigs may be a source of HEV infection for consumers of pig liver and slaughterhouse workers in Brazil.

  17. Detection of DENV-4 genotype I from mosquitoes collected in the city of Manaus, Brazil.

    Science.gov (United States)

    Figueiredo, Mario Luis Garcia de; Alfonso, Helda L; Amarilla, Alberto Anastacio; Figueiredo, Luiz Tadeu Moraes; Aquino, Victor Hugo; Costa, Cristóvão Alves da; Luz, Sergio Luiz Bessa

    2013-02-19

    Dengue epidemics have been reported in Brazil since 1981. In Manaus, a large city in the Amazon region, dengue is endemic with all four-virus serotypes (DENV-1, -2, -3, and -4) simultaneously causing human disease. In 2008, during a surveillance of dengue virus in mosquitoes in the district of Tancredo Neves in Manaus, 260 mosquitoes of Aedes genus were captured, identified and grouped into pools of 10 mosquitoes. RNA extracts of mosquito pools were tested by a RT-Hemi-Nested-PCR for detection of flaviviruses. One amplicon of 222 bp, compatible with dengue virus serotype 4, was obtained from a pool of Aedes aegypti. The nucleotide sequence of the amplicon indicated that the mosquitoes were infected with DENV-4 of genotype I. This virus of Asian origin has been described in Manaus in 2008 infecting acute febrile illness patients. This is the first report of dengue virus serotype 4 genotype I infecting Aedes aegypti in the Americas.

  18. Epidemic history of Hepatitis C virus in Brazil.

    Science.gov (United States)

    Lampe, Elisabeth; Espirito-Santo, Marcia P; Martins, Regina M B; Bello, Gonzalo

    2010-10-01

    Hepatitis C virus (HCV) subtypes 1a, 1b and 3a are the most prevalent strains in Brazil, but very little is known about the epidemic history of these subtypes in the country. A total of 231 HCV NS5B gene sequences (subtype 1a=89, subtype 1b=56, and subtype 3a=86) isolated in Brazil between 1995 and 2007 were analyzed in the present study. Sequences (328-pb) were subjected to phylogenetic analyses and statistical tests of phylogenetic mixing by sampling location and risk group. Our results revealed important variations in the pattern of HCV transmission among subtypes. Transmission of subtype 1a was characterized by dissemination of one major Brazilian lineage with a random virus exchange between different geographical regions but not between IDU and non-IDU populations. Transmission of subtype 1b was characterized by concurrent dissemination of multiple HCV lineages with a restricted virus exchange between country regions and risk groups. Transmission of subtype 3a was characterized by simultaneous spreading of multiple HCV lineages and random phylogenetic mixing by risk group and sampling location. Epidemic histories of major subtypes 1a, 1b and 3a Brazilian clades were estimated using a Bayesian coalescent approach. Our results indicate that all major HCV Brazilian clades probably start to circulate in the country during the second half of the 20th century and displayed roughly similar epidemic histories characterized by an initial phase of exponential expansion and by reduction of growth rates since 1980-1995. This suggests that the expansion of HCV may have been effectively controlled in Brazil. Copyright 2010 Elsevier B.V. All rights reserved.

  19. Dengue virus type 3 in Brazil: a phylogenetic perspective

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    Josélio Maria Galvão de Araújo

    2009-05-01

    Full Text Available Circulation of a new dengue virus (DENV-3 genotype was recently described in Brazil and Colombia, but the precise classification of this genotype has been controversial. Here we perform phylogenetic and nucleotide-distance analyses of the envelope gene, which support the subdivision of DENV-3 strains into five distinct genotypes (GI to GV and confirm the classification of the new South American genotype as GV. The extremely low genetic distances between Brazilian GV strains and the prototype Philippines/L11423 GV strain isolated in 1956 raise important questions regarding the origin of GV in South America.

  20. Genotyping of gastroenteric viruses in hospitalised children: first report of norovirus GII.21 in Brazil

    Directory of Open Access Journals (Sweden)

    Mônica Simões Rocha Ferreira

    2012-12-01

    Full Text Available This retrospective study (April-September 2003 was designed to investigate the roles of the main viruses responsible for cases of acute infantile gastroenteritis in hospitalised children up to two years of age. The viruses were identified in 64.7% (88/136 of the cases and the detection rates of rotavirus A (RVA, norovirus (NoV and astrovirus were 41.9% (57/136, 30.3% (24/79 and 12.7% (7/55, respectively. RVA and NoV were detected in 20 of the 24 reported nosocomial infection cases. This study identified the first circulation of the genotype NoV GII.21 in Brazil and highlights the need to establish differential diagnoses through active laboratorial surveillance.

  1. Detection of hantavirus in bats from remaining rain forest in São Paulo, Brazil.

    Science.gov (United States)

    de Araujo, Jansen; Thomazelli, Luciano Matsumiya; Henriques, Dyana Alves; Lautenschalager, Daniele; Ometto, Tatiana; Dutra, Lilia Mara; Aires, Caroline Cotrin; Favorito, Sandra; Durigon, Edison Luiz

    2012-12-21

    The significant biodiversity found in Brazil is a potential for the emergence of new zoonoses. Study in some places of the world suggest of the presence to hantavirus in tissues of bats. Researches of hantavirus in wildlife, out rodents, are very scarce in Brazil. Therefore we decided to investigate in tissues of different species of wild animals captured in the same region where rodents were detected positive for this virus. The present work analyzed ninety-one animals (64 rodents, 19 opossums, and 8 bats) from a region of the Atlantic forest in Biritiba Mirin City, São Paulo State, Brazil. Lungs and kidneys were used for RNA extraction. The samples were screened for evidence of hantavirus infection by SYBR-Green-based real-time RT-PCR. Sixteen samples positive were encountered among the wild rodents, bats, and opossums. The detection of hantavirus in the lungs and kidneys of three marsupial species (Micoureus paraguayanus, Monodelphis ihering, and Didelphis aurita) as well in two species of bats (Diphylla ecaudata and Anoura caudifer) is of significance because these new hosts could represent an important virus reservoirs. The analysis of nucleotide sequences of the partial S segment revealed that these genes were more related to the Araraquara virus strains. This work reinforces the importance of studying hantavirus in different animal species and performing a continued surveillance before this virus spreads in new hosts and generated serious problems in public health.

  2. Coding Complete Genome for the Mogiana Tick Virus, a Jingmenvirus Isolated from Ticks in Brazil

    Science.gov (United States)

    2017-05-04

    Coding Complete Genome for the Mogiana Tick virus, a Jingmenvirus isolated from ticks in Brazil Erika C Villaa, Sandra R Maruyamab, Isabel KF de...Ribeirão Preto, SP, Brasil Abstract Mogiana tick virus (MGTV) is a segmented Jingmenvirus isolated in 2011 from cattle ticks in Brazil . Here, we...Rhipicephalus microplus) collected from Holstein bulls in Ribeirão Preto, state of São Paulo, Brazil (2). MGTV was one of the earliest

  3. Molecular detection of respiratory viruses: clinical impact

    NARCIS (Netherlands)

    van de Pol, A.C.

    2009-01-01

    Viral respiratory tract infections (LRTIs) cause major morbidity in infants and children. Traditionally, respiratory viruses are detected with conventional tests (viral culture and direct immunofluorescence (DIF)), however nowadays viral diagnostics are being revolutionized by the increased

  4. Zika virus infection in Brazil and human rights obligations.

    Science.gov (United States)

    Diniz, Debora; Gumieri, Sinara; Bevilacqua, Beatriz Galli; Cook, Rebecca J; Dickens, Bernard M

    2017-01-01

    The February 2016 WHO declaration that congenital Zika virus syndrome constitutes a Public Health Emergency of International Concern reacted to the outbreak of the syndrome in Brazil. Public health emergencies can justify a spectrum of human rights responses, but in Brazil, the emergency exposed prevailing inequities in the national healthcare system. The government's urging to contain the syndrome, which is associated with microcephaly among newborns, is confounded by lack of reproductive health services. Women with low incomes in particular have little access to such health services. The emergency also illuminates the harm of restrictive abortion legislation, and the potential violation of human rights regarding women's health and under the UN Conventions on the Rights of the Child and on the Rights of Persons with Disabilities. Suggestions have been proposed by which the government can remedy the widespread healthcare inequities among the national population that are instructive for other countries where congenital Zika virus syndrome is prevalent. © 2016 International Federation of Gynecology and Obstetrics.

  5. Estimated Zika virus importations to Europe by travellers from Brazil

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    Eduardo Massad

    2016-05-01

    Full Text Available Background: Given the interconnectivity of Brazil with the rest of the world, Zika virus (ZIKV infections have the potential to spread rapidly around the world via viremic travellers. The extent of spread depends on the travel volume and the endemicity in the exporting country. In the absence of reliable surveillance data, we did mathematical modelling to estimate the number of importations of ZIKV from Brazil into Europe. Design: We applied a previously developed mathematical model on importations of dengue to estimate the number of ZIKV importations into Europe, based on the travel volume, the probability of being infected at the time of travel, the population size of Brazil, and the estimated incidence of ZIKV infections. Results: Our model estimated between 508 and 1,778 imported infections into Europe in 2016, of which we would expect between 116 and 355 symptomatic Zika infections; with the highest number of importations being into France, Portugal and Italy. Conclusions: Our model identified high-risk countries in Europe. Such data can assist policymakers and public health professionals in estimating the extent of importations in order to prepare for the scale up of laboratory diagnostic assays and estimate the occurrence of Guillain–Barré Syndrome, potential sexual transmission, and infants with congenital ZIKV syndrome.

  6. Parallel evaluation of broad virus detection methods.

    Science.gov (United States)

    Modrof, Jens; Berting, Andreas; Kreil, Thomas R

    2014-01-01

    The testing for adventitious viruses is of critical importance during development and production of biological products. The recent emergence and ongoing development of broad virus detection methods calls for an evaluation of whether these methods can appropriately be implemented into current adventitious agent testing procedures. To assess the suitability of several broad virus detection methods, a comparative experimental study was conducted: four virus preparations, which were spiked at two different concentrations each into two different cell culture media, were sent to four investigators in a blinded fashion for analysis with broad virus detection methods such as polymerase chain reaction-electrospray ionization mass spectrometry (PCR-ESI/MS), microarray, and two approaches utilizing massively parallel sequencing. The results that were reported by the investigators revealed that all methods were able to identify the majority of samples correctly (mean 83%), with a surprisingly narrow range among the methods, that is, between 72% (PCR-ESI/MS) and 95% (microarray). In addition to the correct results, a variety of unexpected assignments were reported for a minority of samples, again with little variation regarding the methods used (range 20-45%), while false negatives were reported for 0-25% of the samples. Regarding assay sensitivity, the viruses were detected by all methods included in this study at concentrations of about 4-5 log10 quantitative PCR copies/mL, and probably with higher sensitivity in some cases. In summary, the broad virus detection methods investigated were shown to be suitable even for detection of relatively low virus concentrations. However, there is also some potential for the production of false-positive as well as false-negative assignments, which indicates the requirement for further improvements before these methods can be considered for routine use. © PDA, Inc. 2014.

  7. Sentinel surveillance of influenza and other respiratory viruses, Brazil, 2000-2010

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    Felipe Teixeira de Mello Freitas

    Full Text Available There are scanty data on the epidemiology of influenza and other respiratory viruses in South America and Brazil. The aim of this study was to summarize the data from the Brazilian surveillance system of influenza and other respiratory viruses and discuss the patterns of viral circulation. The system is based on detecting cases of influenza-like illness in sentinel sites and weekly collection of five nasopharyngeal secretions samples, which are processed in state public health laboratories for respiratory viruses by indirect immunofluorescence assay. Data from 2000 to 2010 were described over time, by region, gender, and age group, and an analysis of Spearman correlation was performed between monthly influenza detection and rainfall and temperature data in two state capitals with the highest number of positive samples, one from the northeast region (Maceió and other from the southern region (Curitiba. There were 3,291,946 visits for influenza-like illness; of these, 37,120 had samples collected and 6421 tested positive: 1690 (26% influenza A, 567 (9% influenza B, 277 (4% parainfluenza 1, 571 (9% parainfluenza 2, 589 (9% parainfluenza 3, 742 (12% adenovirus, and 1985 (31% respiratory syncytial virus. Overall, increased activity of respiratory syncytial virus was observed from March to June, preceding the peak of influenza activity, from May to August, but with regional differences. In Maceió, there was a weak correlation between temperature and influenza detection (ρ = 0.05, but a moderate positive correlation between rainfall and influenza detection (ρ = 0.36. In Curitiba, a high correlation was observed between the decrease in temperature and rainfall and the increase in influenza detection (ρ = -0.83 and -0.78 respectively. These data are important to guide public health control measures as the best time for influenza vaccination and use of antivirals.

  8. MOLECULAR CHARACTERIZATION OF INFLUENZA B VIRUS OUTBREAK ON A CRUISE SHIP IN BRAZIL 2012

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    Samanta Etel Treiger Borborema

    2014-06-01

    Full Text Available In February 2012, an outbreak of respiratory illness occurred on the cruise ship MSC Armonia in Brazil. A 31-year-old female crew member was hospitalized with respiratory failure and subsequently died. To study the etiology of the respiratory illness, tissue taken at necropsy from the deceased woman and respiratory specimens from thirteen passengers and crew members with respiratory symptoms were analyzed. Influenza real-time RT-PCR assays were performed, and the full-length hemagglutinin (HA gene of influenza-positive samples was sequenced. Influenza B virus was detected in samples from seven of the individuals, suggesting that it was the cause of this respiratory illness outbreak. The sequence analysis of the HA gene indicated that the virus was closely related to the B/Brisbane/60/2008-like virus, Victoria lineage, a virus contained in the 2011-12 influenza vaccine for the Southern Hemisphere. Since the recommended composition of the influenza vaccine for use during the 2013 season changed, an intensive surveillance of viruses circulating worldwide is crucial. Molecular analysis is an important tool to characterize the pathogen responsible for an outbreak such as this. In addition, laboratory disease surveillance contributes to the control measures for vaccine-preventable influenza.

  9. An autochthonous case of hepatitis C virus genotype 5a in Brazil: phylogenetic analysis

    DEFF Research Database (Denmark)

    Ribeiro, L.C.; Souto, F.J.D.; do Espirito-Santo, M.P.

    2009-01-01

    Genotype 5 of hepatitis C virus (HCV) has been rarely identified in South America. A female of African descent who never left Brazil was found to be infected by this genotype in Mato Grosso state, Central Brazil. The patient denied drug injections and revealed that she had received blood transfus......Genotype 5 of hepatitis C virus (HCV) has been rarely identified in South America. A female of African descent who never left Brazil was found to be infected by this genotype in Mato Grosso state, Central Brazil. The patient denied drug injections and revealed that she had received blood...

  10. Geographic distribution of hepatitis C virus genotypes in Brazil

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    Campiotto S.

    2005-01-01

    Full Text Available Brazil is a country of continental dimension with a population of different ethnic backgrounds. Thus, a wide variation in the frequencies of hepatitis C virus (HCV genotypes is expected to occur. To address this point, 1,688 sequential samples from chronic HCV patients were analyzed. HCV-RNA was amplified by the RT-PCR from blood samples collected from 1995 to 2000 at different laboratories located in different cities from all Brazilian States. Samples were collected in tubes containing a gel separator, centrifuged in the site of collection and sent by express mail in a refrigerated container to Laboratório Bioquímico Jardim Paulista, São Paulo, SP, Brazil. HCV- RNA was extracted from serum and submitted to RT and nested PCR using standard procedures. Nested PCR products were submitted to cycle sequencing reactions without prior purification. Sequences were analyzed for genotype determination and the following frequencies were found: 64.9% (1,095 for genotype 1, 4.6% (78 for genotype 2, 30.2% (510 for genotype 3, 0.2% (3 for genotype 4, and 0.1% (2 for genotype 5. The frequencies of HCV genotypes were statistically different among Brazilian regions (P = 0.00017. In all regions, genotype 1 was the most frequent (51.7 to 74.1%, reaching the highest value in the North; genotype 2 was more prevalent in the Center-West region (11.4%, especially in Mato Grosso State (25.8%, while genotype 3 was more common in the South (43.2%. Genotypes 4 and 5 were rarely found and only in the Southeast, in São Paulo State. The present data indicate the need for careful epidemiological surveys throughout Brazil since knowing the frequency and distribution of the genotypes would provide key information for understanding the spread of HCV.

  11. Phylogenetic characterization of rabies virus isolates from Carnivora in Brazil.

    Science.gov (United States)

    Kobayashi, Yuki; Inoue, Nana; Sato, Go; Itou, Takuya; Santos, Hamilton P; Brito, Cristina J C; Gomes, Albério A B; Santos, Marli F C; Silva, Marlon V; Mota, Carla S; Ito, Fumio H; Sakai, Takeo

    2007-07-01

    The incidence of canine rabies has been widely reported in Brazil, and new rabies virus (RV) variants, genetically similar to canine RV, have recently been isolated from foxes. In order to derive the epidemiological characteristics of Brazilian Carnivora RV, Brazilian RVs isolated from dogs, cats, and foxes were genetically analyzed. Brazilian Carnivora RV isolates were divided into 2 main lineages. The predominant lineage was found in dogs and cats, which included the Argentinean and Bolivian Carnivora RV isolates, and was extensively distributed throughout Brazil and surrounding countries. The other lineage consisted of three sublineages containing Brazilian dog and fox RV isolates, with the dog sublineages located on an internal branch of 2 fox sublineages, suggesting that RV transmission events might have occurred between foxes and dogs in the past. These results suggest that contact between dogs and wildlife has the potential to generate new rabies variants and that it is important to control RV infection cycles in both dogs and wildlife to prevent spread of rabies infection.

  12. Absence of Bovine leukemia virus (BLV) infection in buffaloes from Amazon and southeast region in Brazil.

    Science.gov (United States)

    De Oliveira, Cairo H S; Resende, Cláudia F; Oliveira, Carlos M C; Barbosa, José D; Fonseca, Antônio A; Leite, Rômulo C; Reis, Jenner K P

    2016-07-01

    Enzootic bovine leucosis is an infectious disease caused by Bovine leukemia virus (BLV) and is well described in bovines. The majority of infected animals are asymptomatic, one to five percent develop lymphoma and from 30 to 50% present a persistent lymphocytosis. The virus occurs naturally in cattle and experimentally in buffaloes, capybaras and rabbits. The occurrence of lymphoma in buffaloes has been attributed to BLV infection by some authors in India and Venezuela, but not confirmed by other studies and little information on natural BLV infection in buffaloes is available. The aim of this study was to evaluate the occurrence of BLV in a sub-sample of buffalo from Amazon and southeast regions in Brazil. Three hundred and fifteen serum samples were negative using commercial AGID and ELISA (ELISA-gp51) which detect anti-BLV glycoprotein gp51 antibodies. The same samples were also evaluated for antibodies to whole virus through a commercial ELISA (ELISA-BLV) in which 77 (24.44%) were found seropositive and two (0.63%) inconclusive. On the other hand, all animals were negative by PCR to BLV targeted to the env and tax genes. These results suggest that ELISA-BLV produces false positive results in buffalo serum (p<0.001). In addition, one buffalo lymphoma sample was negative in both PCR assays used in this study. BLV was not detected in buffaloes from the Amazon basin and the southeast region of Brazil. Serological tests, like ELISA-BLV, usually used for cattle may produce false-positive results for BLV in buffaloes and direct detection tests such as PCR should be chosen in these surveys. The occurrence of lymphoma in buffalo was not associated with BLV infection in the one case analyzed in this work and the etiology and pathogenesis of this disease should be clarified. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. Swine Influenza Virus and Association with the Porcine Respiratory Disease Complex in Pig Farms in Southern Brazil.

    Science.gov (United States)

    Schmidt, C; Cibulski, S P; Andrade, C P; Teixeira, T F; Varela, A P M; Scheffer, C M; Franco, A C; de Almeida, L L; Roehe, P M

    2016-05-01

    Despite the putative endemic status of swine influenza A virus (swIAV) infections, data on the occurrence of swine influenza outbreaks are scarce in Brazil. The aim of this study was to detect and subtype swIAVs from six outbreaks of porcine respiratory disease complex (PRDC) in southern Brazil. Nasal swabs were collected from 66 piglets with signs of respiratory disease in six herds. Lung tissue samples were collected from six necropsied animals. Virus detection was performed by PCR screening and confirmed by virus isolation and hemagglutination (HA). Influenza A subtyping was performed by a real-time reverse transcriptase PCR (rRT-PCR) to detect the A(H1N1)pdm09; other swIAV subtypes were determined by multiplex RT-PCR. In lung tissues, the major bacterial and viral pathogens associated with PRDC (Pasteurella multocida, Mycoplasma hyopneumoniae, Actinobacillus pleuropneumoniae, Haemophilus parasuis and PCV2) were investigated. In some affected pigs, clinico-pathological evaluations were conducted. Influenza A was detected by screening PCR in 46 of 66 swab samples and from five of six lungs. Virus was recovered from pigs of all six herds. Subtype A(H1N1)pdm09 was detected in four of six herds and H1N2 in the other two herds. In lung tissues, further agents involved in PRDC were detected in all cases; Pasteurella multocida was identified in five of six samples and Mycoplasma hyopneumoniae in three of six. Actinobacillus pleuropneumoniae (1/6), Haemophilus parasuis (1/6) and PCV2 (1/6) were also detected. These findings indicate that subtypes A(H1N1)pdm09 and H1N2 were present in pigs in southern Brazil and were associated with PRDC outbreaks. © 2015 Blackwell Verlag GmbH.

  14. Isolation of a phylogenetically distinct rabies virus from a tufted capuchin monkey (Cebus apella) in Brazil.

    Science.gov (United States)

    Kobayashi, Yuki; Sugimoto, Kahori; Mochizuki, Nobuyuki; Segawa, Takao; Itou, Takuya; Carvalho, Adolorata A B; Nociti, Darci P; Mello, Rosane M; Santos, Anna K R A; Ito, Fumio H; Sakai, Takeo

    2013-12-26

    A rabies virus isolate (BRmk1358 strain) was discovered from a rabid tufted capuchin monkey in Brazil. The present study determined the nucleotide sequence of the BRmk1358 strain and compared with the rabies viruses isolated from marmosets and other animals in the Americas. Phylogenetic analyses showed that the BRmk1358 strain formed a lineage distant from that of marmoset rabies virus within the Chiroptera-related rabies virus cluster. This result suggests that the source of rabies infection in the tufted capuchin monkey may have been bat, and that they have a risk to act as rabies reservoir in Brazil. Copyright © 2013 Elsevier B.V. All rights reserved.

  15. Metagenomic analysis reveals Hepatitis A virus in suspected yellow fever cases in Brazil.

    Science.gov (United States)

    Conteville, Liliane C; Filippis, Ana Maria B de; Nogueira, Rita Maria R; Mendonça, Marcos César L de; Vicente, Ana Carolina P

    2018-01-01

    Using a metagenomic approach, we identified hepatitis A virus among cases of acute febrile illnesses that occurred in 2008-2012 in Brazil suspected as yellow fever. These findings reinforce the challenge facing routine clinical diagnosis in complex epidemiological scenarios.

  16. Complete Genome Sequence of a Dengue Virus Serotype 4 Strain Isolated in Roraima, Brazil

    Science.gov (United States)

    Souza, Victor C.; Silva, George A. V.; Maito, Rodrigo M.; Granja, Fabiana; Siqueira, Thalita; Acosta, Pablo O. A.

    2012-01-01

    Dengue is the most important arboviral disease worldwide. We report the complete genome sequence of a dengue virus serotype 4, genotype II strain isolated in 2010 from a patient with classical dengue fever in Boa Vista, Roraima, Brazil. PMID:22247521

  17. Acute gastroenteritis and enteric viruses in hospitalised children in southern Brazil: aetiology, seasonality and clinical outcomes

    Directory of Open Access Journals (Sweden)

    Sonia Maria Raboni

    2014-07-01

    Full Text Available Viral acute gastroenteritis (AG is a significant cause of hospitalisation in children younger than five years. Group A rotavirus (RVA is responsible for 30% of these cases. Following the introduction of RVA immunisation in Brazil in 2006, a decreased circulation of this virus has been observed. However, AG remains an important cause of hospitalisation of paediatric patients and only limited data are available regarding the role of other enteric viruses in these cases. We conducted a prospective study of paediatric patients hospitalised for AG. Stool samples were collected to investigate human adenovirus (HAdV, RVA, norovirus (NoV and astrovirus (AstV. NoV typing was performed by nucleotide sequencing and phylogenetic analysis. From the 225 samples tested, 60 (26% were positive for at least one viral agent. HAdV, NoV, RVA and AstV were detected in 16%, 8%, 6% and 0% of the samples, respectively. Mixed infections were found in nine patients: HAdV/RVA (5, HAdV/NoV (3 and HAdV/NoV/RVA (1. The frequency of fever and lymphocytosis was significantly higher in virus-infected patients. Phylogenetic analysis of NoV indicated that all of these viruses belonged to genotype GII.4. The significant frequency of these pathogens in patients with AG highlights the need to routinely implement laboratory investigations.

  18. Dengue virus type 3 in Rio de Janeiro, Brazil

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    Nogueira Rita Maria R

    2001-01-01

    Full Text Available Dengue virus type 3 was isolated for the first time in the country as an indigenous case from a 40 year-old woman presenting signs and symptoms of a classical dengue fever in the municipality of Nova Iguaçu, State of Rio de Janeiro. This serotype has been associated with dengue haemorrhagic epidemics and the information could be used to implement appropriate prevention and control measures. Virological surveillance was essential in order to detected this new serotype.

  19. Detecting viruses by using salivary diagnostics

    Science.gov (United States)

    Corstjens, Paul L.A.M.; Abrams, William R.; Malamud, Daniel

    2014-01-01

    Background Diagnostics that involve the use of oral fluids have become increasingly available commercially in recent years and are of particular interest because of their relative ease of use, low cost and noninvasive collection of oral fluid for testing. Types of Studies Reviewed The authors discuss the use of salivary diagnostics for virus detection with an emphasis on rapid detection of infection by using point-of-care devices. In particular, they review salivary diagnostics for human immunodeficiency virus, hepatitis C virus and human papillomavirus. Oral mucosal transudate contains secretory immunoglobulin (Ig) A, as well as IgM and IgG, which makes it a good source for immunodiagnostic-based devices. Clinical Implications Because patients often visit a dentist more regularly than they do a physician, there is increased discussion in the dental community regarding the need for practitioners to be aware of salivary diagnostics and to be willing and able to administer these tests to their patients. PMID:23034833

  20. Isolation of Saint Louis Encephalitis Virus from a Horse with Neurological Disease in Brazil

    Science.gov (United States)

    Rosa, Roberta; Costa, Erica Azevedo; Marques, Rafael Elias; Oliveira, Taismara Simas; Furtini, Ronaldo; Bomfim, Maria Rosa Quaresma; Teixeira, Mauro Martins; Paixão, Tatiane Alves; Santos, Renato Lima

    2013-01-01

    St. Louis encephalitis virus (SLEV) is a causative agent of encephalitis in humans in the Western hemisphere. SLEV is a positive-sense RNA virus that belongs to the Flavivirus genus, which includes West Nile encephalitis virus, Japanese encephalitis virus, Dengue virus and other medically important viruses. Recently, we isolated a SLEV strain from the brain of a horse with neurological signs in the countryside of Minas Gerais, Brazil. The SLEV isolation was confirmed by reverse-transcription RT-PCR and sequencing of the E protein gene. Virus identity was also confirmed by indirect immunofluorescence using commercial antibodies against SLEV. To characterize this newly isolated strain in vivo, serial passages in newborn mice were performed and led to hemorrhagic manifestations associated with recruitment of inflammatory cells into the central nervous system of newborns. In summary this is the first isolation of SLEV from a horse with neurological signs in Brazil. PMID:24278489

  1. First report of multiple lineages of dengue viruses type 1 in Rio de Janeiro, Brazil.

    Science.gov (United States)

    dos Santos, Flavia B; Nogueira, Fernanda B; Castro, Márcia G; Nunes, Priscila Cg; de Filippis, Ana Maria B; Faria, Nieli Rc; Simões, Jaqueline Bs; Sampaio, Simone A; Santos, Clarice R; Nogueira, Rita Maria R

    2011-08-03

    In Brazil dengue has been a major public health problem since DENV-1 introduction and spread in 1986. After a low or silent co-circulation, DENV-1 re-emerged in 2009 causing a major epidemic in the country in 2010 and 2011. In this study, the phylogeny of DENV-1 strains isolated in RJ after its first introduction in 1986 and after its emergence in 2009 and 2010 was performed in order to document possible evolutionary patterns or introductions in a re-emergent virus. The analysis of the E gene sequences demonstrated that DENV-1 isolated during 2009/2010 still belong to genotype V (Americas/Africa) but grouping in a distinct clade (lineage II) of that represented by earlier DENV-1 (lineage I). However, strains isolated in 2011 grouped together forming another distinct clade (lineage III). The monitoring of DENV is important to observe the spread of potentially virulent strains as well to evaluate its impact over the population during an outbreak. Whether explosive epidemics reported in Brazil caused mainly by DENV-1 was due to lineage replacement, or due the population susceptibility to this serotype which has not circulated for almost a decade or even due to the occurrence of secondary infections in a hyperendemic country, is not clear. This is the first report of multiple lineages of DENV-1 detected in Brazil.

  2. First report of multiple lineages of dengue viruses type 1 in Rio de Janeiro, Brazil

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    Simões Jaqueline BS

    2011-08-01

    Full Text Available Abstract Background In Brazil dengue has been a major public health problem since DENV-1 introduction and spread in 1986. After a low or silent co-circulation, DENV-1 re-emerged in 2009 causing a major epidemic in the country in 2010 and 2011. In this study, the phylogeny of DENV-1 strains isolated in RJ after its first introduction in 1986 and after its emergence in 2009 and 2010 was performed in order to document possible evolutionary patterns or introductions in a re-emergent virus. Findings The analysis of the E gene sequences demonstrated that DENV-1 isolated during 2009/2010 still belong to genotype V (Americas/Africa but grouping in a distinct clade (lineage II of that represented by earlier DENV-1 (lineage I. However, strains isolated in 2011 grouped together forming another distinct clade (lineage III. Conclusions The monitoring of DENV is important to observe the spread of potentially virulent strains as well to evaluate its impact over the population during an outbreak. Whether explosive epidemics reported in Brazil caused mainly by DENV-1 was due to lineage replacement, or due the population susceptibility to this serotype which has not circulated for almost a decade or even due to the occurrence of secondary infections in a hyperendemic country, is not clear. This is the first report of multiple lineages of DENV-1 detected in Brazil.

  3. Hepatitis E virus in swine and effluent samples from slaughterhouses in Brazil.

    Science.gov (United States)

    dos Santos, Debora Regina Lopes; de Paula, Vanessa Salete; de Oliveira, Jaqueline Mendes; Marchevsky, Renato Sérgio; Pinto, Marcelo Alves

    2011-04-21

    Hepatitis E is an infectious disease which virus (HEV) is highly disseminated in swine herd populations. Sporadic acute human hepatitis E cases have been associated to genotype 3 and 4 strains of HEV also reported in swine populations of endemic and non-endemic areas. With the aim to evaluate the incidence of animals with current infection of HEV, 115 bile samples were collected from three slaughterhouses under inspection by Animal Sanitary Protection Agency of Rio de Janeiro, Brazil. In parallel, effluent samples were collected from six sewage pipe exit sites of two slaughterhouses. HEV RNA was detected in 11 out of 115 (9.6%) bile samples collected and three waste samples from one slaughterhouse. Viral loads observed for bile samples varied from 10(1)-10(5) genome copies/mL and for effluent samples mean load was 10(2) genome copies/mL. Sequencing and phylogenetic analysis classified samples within genotype 3 subtype 3b closely related to the sample obtained from the first reported autochthonous human case and samples from swine of commercial herds in Brazil. Our data demonstrates that although most animals achieve slaughter age (around 20 weeks old) already immune to HEV, a significant number of animals are with current infection at commercial age. Further studies should be addressed to consider risk analysis and possible evaluation of inspection regulations considering food safety measures regarding hepatitis E zoonotic aspect in Brazil. Copyright © 2010 Elsevier B.V. All rights reserved.

  4. Detection of measles, mumps, and rubella viruses.

    Science.gov (United States)

    Tipples, Graham; Hiebert, Joanne

    2011-01-01

    Measles, mumps, and rubella are infections caused by RNA viruses of the same name and are vaccine preventable. The vaccines are frequently administered in a trivalent form. Laboratory diagnostic methods can include indirect detection via antibody (IgM and IgG) detection methods and direct detection by viral culture or viral genome detection. There are challenges for the laboratory in areas with low prevalence due to high vaccine uptake. In those areas, routine serological methods such as IgM detection may have a reduced positive predictive value and thus require confirmation by other methods. Direct detection of viral genomic material using reverse transcription polymerase chain reaction (RT-PCR) methodologies can play an important role for laboratory confirmation of acute infections. Furthermore, genotyping of these three viruses provides useful molecular epidemiological data for differentiating vaccine from wild-type strains, linking cases and outbreaks, and tracking geographic spread and elimination. The purpose of this chapter is to provide guidance for the laboratory diagnosis of measles, mumps, and rubella virus infections. Where assays are commercially available or previously published, the appropriate references are provided as well as brief comments on the interpretation of results. Detailed protocols are provided for the molecular assays which have been developed and more commonly applied in recent years.

  5. The efficiency of concentration methods used to detect enteric viruses in anaerobically digested sludge

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    Tatiana Prado

    2013-02-01

    Full Text Available The presence of enteric viruses in biosolids can be underestimated due to the inefficient methods (mainly molecular methods used to recover the viruses from these matrices. Therefore, the goal of this study was to evaluate the different methods used to recover adenoviruses (AdV, rotavirus species A (RVA, norovirus genogroup II (NoV GII and the hepatitis A virus (HAV from biosolid samples at a large urban wastewater treatment plant in Brazil after they had been treated by mesophilic anaerobic digestion. Quantitative polymerase chain reaction (PCR was used for spiking experiments to compare the detection limits of feasible methods, such as beef extract elution and ultracentrifugation. Tests were performed to detect the inhibition levels and the bacteriophage PP7 was used as an internal control. The results showed that the inhibitors affected the efficiency of the PCR reaction and that beef extract elution is a suitable method for detecting enteric viruses, mainly AdV from biosolid samples. All of the viral groups were detected in the biosolid samples: AdV (90%, RVA, NoV GII (45% and HAV (18%, indicating the viruses' resistance to the anaerobic treatment process. This is the first study in Brazil to detect the presence of RVA, AdV, NoV GII and HAV in anaerobically digested sludge, highlighting the importance of adequate waste management.

  6. Association between microcephaly, Zika virus infection, and other risk factors in Brazil: final report of a case-control study.

    Science.gov (United States)

    de Araújo, Thalia Velho Barreto; Ximenes, Ricardo Arraes de Alencar; Miranda-Filho, Demócrito de Barros; Souza, Wayner Vieira; Montarroyos, Ulisses Ramos; de Melo, Ana Paula Lopes; Valongueiro, Sandra; de Albuquerque, Maria de Fátima Pessoa Militão; Braga, Cynthia; Filho, Sinval Pinto Brandão; Cordeiro, Marli Tenório; Vazquez, Enrique; Cruz, Danielle di Cavalcanti Souza; Henriques, Claudio Maierovitch Pessanha; Bezerra, Luciana Caroline Albuquerque; Castanha, Priscila Mayrelle da Silva; Dhalia, Rafael; Marques-Júnior, Ernesto Torres Azevedo; Martelli, Celina Maria Turchi; Rodrigues, Laura Cunha

    2018-03-01

    A Zika virus epidemic emerged in northeast Brazil in 2015 and was followed by a striking increase in congenital microcephaly cases, triggering a declaration of an international public health emergency. This is the final report of the first case-control study evaluating the potential causes of microcephaly: congenital Zika virus infection, vaccines, and larvicides. The published preliminary report suggested a strong association between microcephaly and congenital Zika virus infection. We did a case-control study in eight public maternity hospitals in Recife, Brazil. Cases were neonates born with microcephaly, defined as a head circumference of 2 SD below the mean. Two controls without microcephaly were matched to each case by expected date of delivery and area of residence. We tested the serum of cases and controls and the CSF of cases for detection of Zika virus genomes with quantitative RT-PCR and for detection of IgM antibodies with capture-IgM ELISA. We also tested maternal serum with plaque reduction neutralisation assays for Zika and dengue viruses. We estimated matched crude and adjusted odds ratios with exact conditional logistic regression to determine the association between microcephaly and Zika virus infection. We screened neonates born between Jan 15 and Nov 30, 2016, and prospectively recruited 91 cases and 173 controls. In 32 (35%) cases, congenital Zika virus infection was confirmed by laboratory tests and no controls had confirmed Zika virus infections. 69 (83%) of 83 cases with known birthweight were small for gestational age, compared with eight (5%) of 173 controls. The overall matched odds ratio was 73·1 (95% CI 13·0-∞) for microcephaly and Zika virus infection after adjustments. Neither vaccination during pregnancy or use of the larvicide pyriproxyfen was associated with microcephaly. Results of laboratory tests for Zika virus and brain imaging results were available for 79 (87%) cases; within these cases, ten were positive for Zika virus

  7. SAINT LOUIS ENCEPHALITIS VIRUS IN MATO GROSSO, CENTRAL-WESTERN BRAZIL

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    Letícia Borges da Silva HEINEN

    2015-06-01

    Full Text Available The dengue virus (DENV, which is frequently involved in large epidemics, and the yellow fever virus (YFV, which is responsible for sporadic sylvatic outbreaks, are considered the most important flaviviruses circulating in Brazil. Because of that, laboratorial diagnosis of acute undifferentiated febrile illness during epidemic periods is frequently directed towards these viruses, which may eventually hinder the detection of other circulating flaviviruses, including the Saint Louis encephalitis virus (SLEV, which is widely dispersed across the Americas. The aim of this study was to conduct a molecular investigation of 11 flaviviruses using 604 serum samples obtained from patients during a large dengue fever outbreak in the state of Mato Grosso (MT between 2011 and 2012. Simultaneously, 3,433 female Culex spp. collected with Nasci aspirators in the city of Cuiabá, MT, in 2013, and allocated to 409 pools containing 1-10 mosquitoes, were also tested by multiplex semi-nested reverse transcription PCR for the same flaviviruses. SLEV was detected in three patients co-infected with DENV-4 from the cities of Cuiabá and Várzea Grande. One of them was a triple co-infection with DENV-1. None of them mentioned recent travel or access to sylvatic/rural regions, indicating that transmission might have occurred within the metropolitan area. Regarding mosquito samples, one pool containing one Culex quinquefasciatus female was positive for SLEV, with a minimum infection rate (MIR of 0.29 per 1000 specimens of this species. Phylogenetic analysis indicates both human and mosquito SLEV cluster, with isolates from genotype V-A obtained from animals in the Amazon region, in the state of Pará. This is the first report of SLEV molecular identification in MT.

  8. Feline immunodeficiency virus and feline leukemia virus: frequency and associated factors in cats in northeastern Brazil.

    Science.gov (United States)

    Lacerda, L C; Silva, A N; Freitas, J S; Cruz, R D S; Said, R A; Munhoz, A D

    2017-05-10

    Our aims were to determine the frequencies of feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) in owned and stray cats in the northeastern region of Brazil, ascertain the status of FeLV infection, and investigate potential associated factors among the owned cats. Blood samples from 200 asymptomatic owned cats and 30 stray cats were processed using nested PCR and commercial immunochromatographic tests to diagnose infections. To evaluate the factors associated with FIV and/or FeLV in owned cats, a semi-structured interview was conducted with each owner about the animal's environment, and these data were subjected to unconditional logistic regression. The frequencies for owned cats were 6% (12/200) and 3% (6/200) for FIV and FeLV, respectively. No owned cat was positive for both viruses. Stray cats showed frequencies of 6.66% (2/30) and 0% (0/30) for FIV and FeLV, respectively. Contact with other cats and living in peri-urban areas were considered to be risk factors (P < 0.05) for FIV. We did not identify any factors associated with infections with FeLV. Our results confirm the presence of these two retroviruses in the region under study. Our use of different diagnostic techniques allowed us to determine the frequency of retroviruses in the feline population more accurately, particularly with regard to infections by FeLV, which have complex pathogenesis.

  9. Detection of Aichi virus in South Korea.

    Science.gov (United States)

    Han, Tae-Hee; Park, Sang Hun; Hwang, Eung-Soo; Reuter, Gabor; Chung, Ju-Young

    2014-07-01

    Aichi virus (AiV) is considered to be a possible etiologic agent of acute gastroenteritis (GE). We analyzed 1,568 stool samples collected by the Seoul Metropolitan Health Research Center from patients with GE during outbreaks in Seoul, together with 378 archived common-enteric-virus-negative stool samples from children with GE hospitalized at a tertiary hospital in Seoul. AiV was detected in 1.7 % (27/1,568) of the first group but not found in the second group (0 %, 0/378). Genotypes A and B of AiV were both detected in this study. This is the first study confirming the circulation of AiV in Korea.

  10. Severity of tegumentary leishmaniasis is not exclusively associated with Leishmania RNA virus 1 infection in Brazil

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    Luiza de Oliveira Ramos Pereira

    2013-08-01

    Full Text Available Leishmania RNA virus (LRV has been shown to be a symbiotic component of Leishmania parasites in South America. Nested retro-transcription polymerase chain reaction was employed to investigate LRV1 presence in leishmaniasis lesions from Brazil. In endemic areas of Rio de Janeiro (RJ, no LRV1 infection was observed even with mucosal involvement. LRV1 was only detected in Leishmania (V. guyanensis cutaneous lesions from the northern region, which were obtained from patients presenting with disease reactivation after clinical cure of their primary lesions. Our results indicated that the severity of leishmaniasis in some areas of RJ, where Leishmania (V. brazi-liensis is the primary etiological agent, was not associated with Leishmania LRV1 infection.

  11. Detection of virus in shrimp using digital color correlation

    Science.gov (United States)

    Alvarez-Borrego, Josue; Chavez-Sanchez, Cristina; Bueno-Ibarra, Mario A.

    1999-07-01

    Detection of virus in shrimp tissue using digital color correlation is presented. Phase filters in three channels (red, green and blue) were used in order to detect HPV virus like target. These first results obtained showed that is possible to detect virus in shrimp tissue. More research must be made with color correlation in order to consider natural morphology of the virus, color, scale and rotation and noise in the samples.

  12. Detection of three honeybee viruses simultaneously by a single ...

    African Journals Online (AJOL)

    A single multiplex reverse transcriptase (RT) polymerase chain reaction (PCR) assay was developed for the simultaneous detection of three honeybee viruses: acute bee paralysis virus (ABPV), sacbrood virus (SBV) and black queen cell virus (BQCV). Unique PCR primers were designed from the complete genome ...

  13. Detection, isolation, and persistence of viruses within bivalve mollusks

    Science.gov (United States)

    Norovirus (NV), hepatitis A virus (HAV), and other virus transmission by molluscan shellfish is a significant issue. Research at the ARS-Dover DE laboratory has led to the development of improved methods for detecting these viruses. To identify pathogenic viruses within mollusks, a rapid highly-se...

  14. Isolation of Infective Zika Virus from Urine and Saliva of Patients in Brazil.

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    Myrna C Bonaldo

    2016-06-01

    Full Text Available Zika virus (ZIKV is an emergent threat provoking a worldwide explosive outbreak. Since January 2015, 41 countries reported autochthonous cases. In Brazil, an increase in Guillain-Barré syndrome and microcephaly cases was linked to ZIKV infections. A recent report describing low experimental transmission efficiency of its main putative vector, Ae. aegypti, in conjunction with apparent sexual transmission notifications, prompted the investigation of other potential sources of viral dissemination. Urine and saliva have been previously established as useful tools in ZIKV diagnosis. Here, we described the presence and isolation of infectious ZIKV particles from saliva and urine of acute phase patients in the Rio de Janeiro state, Brazil.Nine urine and five saliva samples from nine patients from Rio de Janeiro presenting rash and other typical Zika acute phase symptoms were inoculated in Vero cell culture and submitted to specific ZIKV RNA detection and quantification through, respectively, NAT-Zika, RT-PCR and RT-qPCR. Two ZIKV isolates were achieved, one from urine and one from saliva specimens. ZIKV nucleic acid was identified by all methods in four patients. Whenever both urine and saliva samples were available from the same patient, urine viral loads were higher, corroborating the general sense that it is a better source for ZIKV molecular diagnostic. In spite of this, from the two isolated strains, each from one patient, only one derived from urine, suggesting that other factors, like the acidic nature of this fluid, might interfere with virion infectivity. The complete genome of both ZIKV isolates was obtained. Phylogenetic analysis revealed similarity with strains previously isolated during the South America outbreak.The detection of infectious ZIKV particles in urine and saliva of patients during the acute phase may represent a critical factor in the spread of virus. The epidemiological relevance of this finding, regarding the contribution

  15. Isolation of Infective Zika Virus from Urine and Saliva of Patients in Brazil.

    Science.gov (United States)

    Bonaldo, Myrna C; Ribeiro, Ieda P; Lima, Noemia S; Dos Santos, Alexandre A C; Menezes, Lidiane S R; da Cruz, Stephanie O D; de Mello, Iasmim S; Furtado, Nathália D; de Moura, Elaine E; Damasceno, Luana; da Silva, Kely A B; de Castro, Marcia G; Gerber, Alexandra L; de Almeida, Luiz G P; Lourenço-de-Oliveira, Ricardo; Vasconcelos, Ana Tereza R; Brasil, Patrícia

    2016-06-01

    Zika virus (ZIKV) is an emergent threat provoking a worldwide explosive outbreak. Since January 2015, 41 countries reported autochthonous cases. In Brazil, an increase in Guillain-Barré syndrome and microcephaly cases was linked to ZIKV infections. A recent report describing low experimental transmission efficiency of its main putative vector, Ae. aegypti, in conjunction with apparent sexual transmission notifications, prompted the investigation of other potential sources of viral dissemination. Urine and saliva have been previously established as useful tools in ZIKV diagnosis. Here, we described the presence and isolation of infectious ZIKV particles from saliva and urine of acute phase patients in the Rio de Janeiro state, Brazil. Nine urine and five saliva samples from nine patients from Rio de Janeiro presenting rash and other typical Zika acute phase symptoms were inoculated in Vero cell culture and submitted to specific ZIKV RNA detection and quantification through, respectively, NAT-Zika, RT-PCR and RT-qPCR. Two ZIKV isolates were achieved, one from urine and one from saliva specimens. ZIKV nucleic acid was identified by all methods in four patients. Whenever both urine and saliva samples were available from the same patient, urine viral loads were higher, corroborating the general sense that it is a better source for ZIKV molecular diagnostic. In spite of this, from the two isolated strains, each from one patient, only one derived from urine, suggesting that other factors, like the acidic nature of this fluid, might interfere with virion infectivity. The complete genome of both ZIKV isolates was obtained. Phylogenetic analysis revealed similarity with strains previously isolated during the South America outbreak. The detection of infectious ZIKV particles in urine and saliva of patients during the acute phase may represent a critical factor in the spread of virus. The epidemiological relevance of this finding, regarding the contribution of alternative non

  16. Isolation of Infective Zika Virus from Urine and Saliva of Patients in Brazil

    Science.gov (United States)

    da Silva, Kely A. B.; de Castro, Marcia G.; Gerber, Alexandra L.; de Almeida, Luiz G. P.; Lourenço-de-Oliveira, Ricardo; Vasconcelos, Ana Tereza R.

    2016-01-01

    Background Zika virus (ZIKV) is an emergent threat provoking a worldwide explosive outbreak. Since January 2015, 41 countries reported autochthonous cases. In Brazil, an increase in Guillain-Barré syndrome and microcephaly cases was linked to ZIKV infections. A recent report describing low experimental transmission efficiency of its main putative vector, Ae. aegypti, in conjunction with apparent sexual transmission notifications, prompted the investigation of other potential sources of viral dissemination. Urine and saliva have been previously established as useful tools in ZIKV diagnosis. Here, we described the presence and isolation of infectious ZIKV particles from saliva and urine of acute phase patients in the Rio de Janeiro state, Brazil. Methodology/Principal Findings Nine urine and five saliva samples from nine patients from Rio de Janeiro presenting rash and other typical Zika acute phase symptoms were inoculated in Vero cell culture and submitted to specific ZIKV RNA detection and quantification through, respectively, NAT-Zika, RT-PCR and RT-qPCR. Two ZIKV isolates were achieved, one from urine and one from saliva specimens. ZIKV nucleic acid was identified by all methods in four patients. Whenever both urine and saliva samples were available from the same patient, urine viral loads were higher, corroborating the general sense that it is a better source for ZIKV molecular diagnostic. In spite of this, from the two isolated strains, each from one patient, only one derived from urine, suggesting that other factors, like the acidic nature of this fluid, might interfere with virion infectivity. The complete genome of both ZIKV isolates was obtained. Phylogenetic analysis revealed similarity with strains previously isolated during the South America outbreak. Conclusions/Significance The detection of infectious ZIKV particles in urine and saliva of patients during the acute phase may represent a critical factor in the spread of virus. The epidemiological

  17. The Unknown Computer Viruses Detection Based on Similarity

    Science.gov (United States)

    Liu, Zhongda; Nakaya, Naoshi; Koui, Yuuji

    New computer viruses are continually being generated and they cause damage all over the world. In general, current anti-virus software detects viruses by matching a pattern based on the signature; thus, unknown viruses without any signature cannot be detected. Although there are some static analysis technologies that do not depend on signatures, virus writers often use code obfuscation techniques, which make it difficult to execute a code analysis. As is generally known, unknown viruses and known viruses share a common feature. In this paper we propose a new static analysis technology that can circumvent code obfuscation to extract the common feature and detect unknown viruses based on similarity. The results of evaluation experiments demonstrated that this technique is able to detect unknown viruses without false positives.

  18. Real-Time Detection of a Virus Using Detection Dogs

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    Craig eAngle

    2016-01-01

    Full Text Available Viral infections are ubiquitous in humans, animals, and plants. Real-time methods to identify viral infections are limited and do not exist for use in harsh or resource-constrained environments. Previous research identified that tissues produce unique volatile organic compounds (VOC and demonstrated that VOC concentrations change during pathologic states including infection, neoplasia, or metabolic disease. Patterns of VOC expression may be pathogen-specific and may be associated with an odor that could be used for disease detection.We investigated the ability of two trained dogs to detect cell cultures infected with bovine viral diarrhea virus (BVDV and to discriminate BVDV-infected cell cultures from uninfected cell cultures and from cell cultures infected with bovine herpes virus 1 (BHV 1 and bovine parainfluenza virus 3 (BPIV 3. Dogs were trained to recognize cell cultures infected with two different biotypes of BVDV propagated in MDBK cells using one of three culture media. For detection trials, one target and seven distractors were presented on a scent wheel by a dog handler unaware of the location of targets and distractors.Detection of BVDV- infected cell cultures by Dog 1 had a diagnostic sensitivity of 0.850 (95% CI: 0.701 - 0.942, which was lower than Dog 2 (0.967, 95% CI: 0.837 - 0.994. Both dogs exhibited very high diagnostic specificity (0.981, 95% CI: 0.960 - 0.993 and (0.993, 95% CI: 0.975 - 0.999, respectively.These findings demonstrate that trained dogs can differentiate between cultured cells infected with BVDV, BHV1, and BPIV3 and are a realistic real-time mobile pathogen sensing technology for viral pathogens. The ability to discriminate between target and distractor samples plausibly results from expression of unique VOC patterns virus-infected and uninfected cells.

  19. Rapid Detection of the Varicella Zoster Virus

    Science.gov (United States)

    Lewis, Michelle P.; Harding, Robert

    2011-01-01

    1.Technology Description-Researchers discovered that when the Varicella Zoster Virus (VZV) reactivates from latency in the body, the virus is consistently present in saliva before the appearance of skin lesions. A small saliva sample is mixed with a specialized reagent in a test kit. If the virus is present in the saliva sample, the mixture turns a red color. The sensitivity and specificity emanates from an antibody-antigen reaction. This technology is a rapid, non-invasive, point of-of-care testing kit for detecting the virus from a saliva sample. The device is easy to use and can be used in clinics and in remote locations to quickly detect VZV and begin treatment with antiviral drugs. 2.Market Opportunity- RST Bioscience will be the first and only company to market a rapid, same day test kit for the detection of VZV in saliva. The RST detection test kit will have several advantages over existing, competitive technology. The test kit is self contained and laboratory equipment is not required for analysis of the sample. Only a single saliva sample is required to be taken instead of blood or cerebral spinal fluid. The test kit is portable, sterile and disposable after use. RST detection test kits require no electrical power or expensive storage equipment and can be used in remote locations. 3.Market Analysis- According to the CDC, it is estimated that 1 million cases of shingles occur each year in the U.S. with more than half over the age of sixty. There is a high demand for rapid diagnostics by the public. The point-of-care testing (POCT) market is growing faster than other segments of in vitro diagnostics. According to a July 2007 InteLab Corporation industry report the overall market for POCT was forecast to increase from $10.3 billion in 2005 to $18.7 billion by 2011. The market value of this test kit has not been determined. 4.Competition- The VZV vaccine prevents 50% of cases and reduces neuralgia by 66%. The most popular test detects VZV-specific IgM antibody

  20. Genome analysis of yellow fever virus of the ongoing outbreak in Brazil reveals polymorphisms

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    Myrna C Bonaldo

    Full Text Available The current yellow fever outbreak in Brazil is the most severe one in the country in recent times. It has rapidly spread to areas where YF virus (YFV activity has not been observed for more than 70 years and vaccine coverage is almost null. Here, we sequenced the whole YFV genome of two naturally infected howler-monkeys (Alouatta clamitans obtained from the Municipality of Domingos Martins, state of Espírito Santo, Brazil. These two ongoing-outbreak genome sequences are identical. They clustered in the 1E sub-clade (South America genotype I along with the Brazilian and Venezuelan strains recently characterised from infections in humans and non-human primates that have been described in the last 20 years. However, we detected eight unique amino acid changes in the viral proteins, including the structural capsid protein (one change, and the components of the viral replicase complex, the NS3 (two changes and NS5 (five changes proteins, that could impact the capacity of viral infection in vertebrate and/or invertebrate hosts and spreading of the ongoing outbreak.

  1. Estimated global exportations of Zika virus infections via travellers from Brazil from 2014 to 2015.

    Science.gov (United States)

    Quam, Mikkel B; Wilder-Smith, Annelies

    2016-06-01

    The ongoing Zika pandemic in Latin America illustrates a potential source for further globalized spread. Here, we assessed global travel-related Zika virus exportations from Brazil during the initial year of the epidemic. Similar to subsequent national notifications, we estimated 584-1786 exported Zika cases from Brazil occurred September 2014-August 2015. © International Society of Travel Medicine, 2016. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  2. The Zika Virus Epidemic in Brazil: From Discovery to Future Implications

    Directory of Open Access Journals (Sweden)

    Rachel Lowe

    2018-01-01

    Full Text Available The first confirmed case of Zika virus infection in the Americas was reported in Northeast Brazil in May 2015, although phylogenetic studies indicate virus introduction as early as 2013. Zika rapidly spread across Brazil and to more than 50 other countries and territories on the American continent. The Aedes aegypti mosquito is thought to be the principal vector responsible for the widespread transmission of the virus. However, sexual transmission has also been reported. The explosively emerging epidemic has had diverse impacts on population health, coinciding with cases of Guillain–Barré Syndrome and an unexpected epidemic of newborns with microcephaly and other neurological impairments. This led to Brazil declaring a national public health emergency in November 2015, followed by a similar decision by the World Health Organization three months later. While dengue virus serotypes took several decades to spread across Brazil, the Zika virus epidemic diffused within months, extending beyond the area of permanent dengue transmission, which is bound by a climatic barrier in the south and low population density areas in the north. This rapid spread was probably due to a combination of factors, including a massive susceptible population, climatic conditions conducive for the mosquito vector, alternative non-vector transmission, and a highly mobile population. The epidemic has since subsided, but many unanswered questions remain. In this article, we provide an overview of the discovery of Zika virus in Brazil, including its emergence and spread, epidemiological surveillance, vector and non-vector transmission routes, clinical complications, and socio-economic impacts. We discuss gaps in the knowledge and the challenges ahead to anticipate, prevent, and control emerging and re-emerging epidemics of arboviruses in Brazil and worldwide.

  3. Persistence of the tissue culture origin vaccine for infectious laryngotracheitis virus in commercial chicken flocks in Brazil.

    Science.gov (United States)

    Parra, Silvana H Santander; Nuñez, Luis F; Astolfi-Ferreira, Claudete S; Ferreira, Antonio J Piantino

    2015-11-01

    Infectious laryngotracheitis (ILT) is a respiratory disease of great importance that causes serious economic losses in the poultry industry. Its control is based on biosecurity procedures and vaccination programs that use live attenuated vaccines such as tissue culture origin (TCO), chicken embryo origin (CEO), and vectored vaccines. However, problems have been reported, such as the reversion of virulence, virus latency, and field virus outbreaks. Several molecular techniques have been developed to differentiate between the field and vaccine strains. This study was conducted to determine the presence of infectious laryngotracheitis virus (ILTV) in Brazil from 2012 to 2014. PCR-RFLP (restriction fragment length polymorphism) was used to detect and differentiate ILTV strains; DNA sequencing and predictive RFLP analysis were also used for this purpose. Molecular analysis detected the presence of ILTV in 15 samples that were characterized as strains of TCO vaccine origin. This study showed that the ILTV TCO vaccine strain has been circulating in commercial chicken flocks in Brazil since its introduction during the 2002 outbreak. © 2015 Poultry Science Association Inc.

  4. INTERMEDIATE ENDEMICITY OF HEPATITIS A VIRUS INFECTION IN RURAL SETTLEMENT PROJECTS OF SOUTHWEST GOIÁS, BRAZIL.

    Science.gov (United States)

    Pinheiro, Raquel Silva; Araújo, Lyriane Apolinário de; Caetano, Karlla Antonieta Amorim; Matos, Marcos André de; Carneiro, Megmar Aparecida dos Santos; Teles, Sheila Araújo

    2015-01-01

    Rural populations present an elevated risk of exposure to hepatitis A virus. The objective of this study was to estimate the prevalence and risk factors associated with hepatitis A virus infection among residents of rural settlement projects of southwest Goiás, Central Brazil. A total of 466 residents were interviewed and tested for the detection of anti- hepatitis A virus antibodies by ELISA. The global prevalence of anti- hepatitis A virus was 82.2%. In individuals aged 5-9 years and 10-19 years, the prevalence was 15% and 58.8%, respectively. Persons in the 10-19 age group, with a history of life in encampments, with more than five people per residence consuming well water, were predictors for exposure to hepatitis A virus. Our results suggest that the hepatitis A virus endemicity in rural settlements in southwest Goiás similar to that found in the urban population of the Midwest Region, confirming the implementation of universal hepatitis A vaccination in children.

  5. Extreme Water Deficit in Brazil Detected from Space

    Science.gov (United States)

    Vieira Getirana

    2016-01-01

    Extreme droughts have caused significant socioeconomic and environmental damage worldwide. In Brazil, ineffective energy development and water management policies have magnified the impacts of recent severe droughts, which include massive agricultural losses, water supply restrictions, and energy rationing. Spaceborne remote sensing data advance our understanding of the spatiotemporal variability of large-scale droughts and enhance the detection and monitoring of extreme water-related events. In this study, data derived from the Gravity Recovery and Climate Experiment (GRACE) mission are used to detect and quantify an extended major drought over eastern Brazil and provide estimates of impacted areas and region-specific water deficits. Two structural breakpoint detection methods were applied to time series of GRACE-based terrestrial water storage anomalies (TWSA), determining when two abrupt changes occurred. One, in particular, defines the beginning of the current drought. Using TWSA, a water loss rate of 26.1 cmyr21 over southeastern Brazil was detected from 2012 to 2015. Based on analysis of Global Land Data Assimilation System(GLDAS) outputs, the extreme drought is mostly related to lower-than-usual precipitation rates, resulting in high soil moisture depletion and lower-than-usual rates of evapotranspiration. A reduction of 2023 of precipitation over an extended period of 3 years is enough to raise serious water scarcity conditions in the country. Correlations between monthly time series of both grid-based TWSA and ground-based water storage measurements at 16 reservoirs located within southeastern Brazil varied from 0.42 to 0.82. Differences are mainly explained by reservoir sizes and proximity to the drought nucleus.

  6. Human vaccinia-like virus outbreaks in São Paulo and Goiás States, Brazil: virus detection, isolation and identification Surtos de vírus Vaccinia-like nos Estados de São Paulo e Goiás, Brasil: detecção, isolamento e identificação viral

    Directory of Open Access Journals (Sweden)

    Teresa Keico Nagasse-Sugahara

    2004-04-01

    Full Text Available Since October 2001, the Adolfo Lutz Institute has been receiving vesicular fluids and scab specimens of patients from Paraíba Valley region in the São Paulo and Minas Gerais States and from São Patricio Valley, in the Goiás State. Epidemiological data suggested that the outbreaks were caused by Cowpox virus or Vaccinia virus. Most of the patients are dairy milkers that had vesiculo-pustular lesions on the hands, arms, forearms, and some of them, on the face. Virus particles with orthopoxvirus morphology were detected by direct electron microscopy (DEM in samples of 49 (66.21% patients of a total of 74 analyzed. Viruses were isolated in Vero cell culture and on chorioallantoic membrane (CAM of embryonated chicken eggs. Among 21 samples submitted to PCR using primers for hemagglutinin (HA gene, 19 were positive. Restriction digestion with TaqI resulted in four characteristic Vaccinia virus fragments. HA nucleotide sequences showed 99.9% similarity with Cantagalo virus, described as a strain of Vaccinia virus. The only difference observed was the substitution of one nucleotide in the position 616 leading to change in one amino acid of the protein in the position 206. The phylogenetic analysis showed that the isolates clustered together with Cantagalo virus, other Vaccinia strains and Rabbitpox virus.A partir de outubro de 2001, o Instituto Adolfo Lutz tem recebido amostras de líquido vesicular e crostas de lesões de pele de pacientes das regiões do Vale do Paraíba, Estado de São Paulo e do Vale do São Patricio, Estado de Goiás. Os dados clínicos e epidemiológicos sugeriam que os surtos poderiam ser causados por Cowpox virus ou Vaccinia virus. A maioria dos pacientes era ordenhadores que tinham lesões vesicopustulares nas mãos, braços, antebraços e alguns na face. A análise por microscopia eletrônica direta (MED detectou partículas com morfologia de vírus do gênero Orthopoxvirus em amostras de 49 (66,21% pacientes dos 74

  7. Enteric viruses in HIV-1 seropositive and HIV-1 seronegative children with diarrheal diseases in Brazil

    Science.gov (United States)

    Rocha, Monica Simões; Fumian, Tulio Machado; Maranhão, Adriana Gonçalves; de Assis, Rosane Maria; Xavier, Maria da Penha Trindade Pinheiro; Rocha, Myrna Santos; Miagostovich, Marize Pereira; Leite, José Paulo Gagliardi; Volotão, Eduardo de Mello

    2017-01-01

    Diarrheal diseases (DD) have distinct etiological profiles in immune-deficient and immune-competent patients. This study compares detection rates, genotype distribution and viral loads of different enteric viral agents in HIV-1 seropositive (n = 200) and HIV-1 seronegative (n = 125) children hospitalized with DD in Rio de Janeiro, Brazil. Except for group A rotavirus (RVA), which were detected through enzyme immunoassay, the other enteric viruses (norovirus [NoV], astrovirus [HAstV], adenovirus [HAdV] and bocavirus [HBoV]) were detected through PCR or RT-PCR. A quantitative PCR was performed for RVA, NoV, HAstV, HAdV and HBoV. Infections with NoV (19% vs. 9.6%; p<0.001), HBoV (14% vs. 7.2%; p = 0.042) and HAdV (30.5% vs. 14.4%; p<0.001) were significantly more frequent among HIV-1 seropositive children. RVA was significantly less frequent among HIV-1 seropositive patients (6.5% vs. 20%; p<0.001). Similarly, frequency of infection with HAstV was lower among HIV-1 seropositive children (5.5% vs. 12.8%; p = 0.018). Among HIV-1 seropositive children 33 (16.5%) had co-infections, including three enteric viruses, such as NoV, HBoV and HAdV (n = 2) and NoV, HAstV and HAdV (n = 2). The frequency of infection with more than one virus was 17 (13.6%) in the HIV-1 negative group, triple infection (NoV + HAstV + HBoV) being observed in only one patient. The median viral load of HAstV in feces was significantly higher among HIV-1 positive children compared to HIV-1 negative children. Concerning children infected with RVA, NoV, HBoV and HAdV, no statistically significant differences were observed in the medians of viral loads in feces, comparing HIV-1 seropositive and HIV-1 seronegative children. Similar detection rates were observed for RVA, HAstV and HAdV, whilst NoV and HBoV were significantly more prevalent among children with CD4+ T lymphocyte count below 200 cells/mm3. Enteric viruses should be considered an important cause of DD in HIV-1 seropositive children, along with

  8. Enteric viruses in HIV-1 seropositive and HIV-1 seronegative children with diarrheal diseases in Brazil.

    Directory of Open Access Journals (Sweden)

    Silvana Augusta Rodrigues Portes

    Full Text Available Diarrheal diseases (DD have distinct etiological profiles in immune-deficient and immune-competent patients. This study compares detection rates, genotype distribution and viral loads of different enteric viral agents in HIV-1 seropositive (n = 200 and HIV-1 seronegative (n = 125 children hospitalized with DD in Rio de Janeiro, Brazil. Except for group A rotavirus (RVA, which were detected through enzyme immunoassay, the other enteric viruses (norovirus [NoV], astrovirus [HAstV], adenovirus [HAdV] and bocavirus [HBoV] were detected through PCR or RT-PCR. A quantitative PCR was performed for RVA, NoV, HAstV, HAdV and HBoV. Infections with NoV (19% vs. 9.6%; p<0.001, HBoV (14% vs. 7.2%; p = 0.042 and HAdV (30.5% vs. 14.4%; p<0.001 were significantly more frequent among HIV-1 seropositive children. RVA was significantly less frequent among HIV-1 seropositive patients (6.5% vs. 20%; p<0.001. Similarly, frequency of infection with HAstV was lower among HIV-1 seropositive children (5.5% vs. 12.8%; p = 0.018. Among HIV-1 seropositive children 33 (16.5% had co-infections, including three enteric viruses, such as NoV, HBoV and HAdV (n = 2 and NoV, HAstV and HAdV (n = 2. The frequency of infection with more than one virus was 17 (13.6% in the HIV-1 negative group, triple infection (NoV + HAstV + HBoV being observed in only one patient. The median viral load of HAstV in feces was significantly higher among HIV-1 positive children compared to HIV-1 negative children. Concerning children infected with RVA, NoV, HBoV and HAdV, no statistically significant differences were observed in the medians of viral loads in feces, comparing HIV-1 seropositive and HIV-1 seronegative children. Similar detection rates were observed for RVA, HAstV and HAdV, whilst NoV and HBoV were significantly more prevalent among children with CD4+ T lymphocyte count below 200 cells/mm3. Enteric viruses should be considered an important cause of DD in HIV-1 seropositive children, along

  9. Bovine respiratory disease associated with Histophilus somni and bovine respiratory syncytial virus in a beef cattle feedlot from Southeastern Brazil

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    Selwyn Arligton Headley

    2017-03-01

    Full Text Available Bovine respiratory disease (BRD is a complex multifactorial and multi-etiological disease entity that is responsible for the morbidity and mortality particularly in feedlot cattle from North America. Information relative to the occurrence of BRD in Brazil and the associated infectious agents are lacking. This study investigated the participation of infectious agents of BRD in a beef cattle feedlot from Southeastern Brazil. Nasopharyngeal swabs of 11% (10/90 of cattle (n, 450 with clinical manifestations of respiratory distress were analyzed by targeting specific genes of the principal infectious pathogens of BRD. In addition, pulmonary fragments of one the animals that died were collected for histopathological and molecular diagnoses. The nucleic acids of Histophilus somni and bovine respiratory syncytial virus (BRSV were identified in 20% (2/10 of the nasopharyngeal swabs of the animals with respiratory distress; another contained only BRSV RNA. Moreover, the nucleic acids of both infectious agents were amplified from the pulmonary fragments of the animal that died with histopathological evidence of bronchopneumonia and interstitial pneumonia; the nasopharyngeal swab of this animal also contained the nucleic acids of both pathogens. Additionally, all PCR and/or RT-PCR assays designed to detect the specific genes of Mannheimia haemolytica, Pasteurella multocida, Mycoplasma bovis, bovine viral diarrhea virus, bovine herpesvirus -1, bovine parainfluenza virus-3, and bovine coronavirus yielded negative results. Phylogenetic analyses suggest that the isolates of H. somni circulating in Brazil are similar to those identified elsewhere, while there seem to be diversity between the isolates of BRSV within cattle herds from different geographical locations of Brazil.

  10. Aedes albopictus may not be vector of dengue virus in human epidemics in Brazil

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    Degallier Nicolas

    2003-01-01

    Full Text Available Over 60,500 dengue cases were reported in the state of Espírito Santo (ES, Brazil, between 1995 and 1998. The study's purpose was to identify whether Aedes albopictus was transmitting the dengue virus during an epidemic in the locality of Vila Bethânia (Viana County,Vitória, ES. From April 3 to 9, 1998, blood and serum samples were collected daily for virus isolation and serological testing. Four autochthonous cases were confirmed through DEN 1 virus isolation and two autochthonous cases through MAC ELISA testing. Of 37 Ae. aegypti and 200 Ae. albopictus adult mosquitoes collected and inoculated, DEN1 virus was isolated only from a pool of two Ae. aegypti female mosquitoes. The study results suggest that Ae. albopictus still cannot be considered an inter-human vector in dengue epidemics in Brazil.

  11. Respiratory syncytial virus groups A and B in Porto Alegre, Brazil, from 1990 to 1995 and 1998

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    Straliotto Selir M

    2001-01-01

    Full Text Available We analyzed the respiratory syncytial virus (RSV groups and their epidemiological pattern that were detected over the course of seven years in southern Brazil. The two RSV groups co-circulated each year, but frequencies of groups A and B varied both between and within yearly outbreaks. In 1991, group A predominated over group B (p=0.0016. RSV outbreaks analyzed showed a temperature-dependent pattern and no association with rainfall, similarly to other countries from southern South America. Knowledge of the variants is important in terms of both diagnosis and definition of a vaccine composition.

  12. Recurrent Potent Human Neutralizing Antibodies to Zika Virus in Brazil and Mexico

    OpenAIRE

    Robbiani, Davide F.; Khouri, Ricardo; Gristick, Harry B.; Lee, Yu E.; West, Anthony P.; Bjorkman, Pamela J.

    2017-01-01

    Antibodies to Zika virus (ZIKV) can be protective. To examine the antibody response in individuals who develop high titers of anti-ZIKV antibodies, we screened cohorts in Brazil and Mexico for ZIKV envelope domain III (ZEDIII) binding and neutralization. We find that serologic reactivity to dengue 1 virus (DENV1) EDIII before ZIKV exposure is associated with increased ZIKV neutralizing titers after exposure. Antibody cloning shows that donors with high ZIKV neutralizing antibody titers have e...

  13. Bovine leukaemia virus genotypes 5 and 6 are circulating in cattle from the state of São Paulo, Brazil.

    Science.gov (United States)

    Gregory, Lilian; Carrillo Gaeta, Natália; Araújo, Jansen; Matsumiya Thomazelli, Luciano; Harakawa, Ricardo; Ikuno, Alice A; Hiromi Okuda, Liria; de Stefano, Eliana; Pituco, Edviges Maristela

    2017-12-01

    Enzootic bovine leucosis (EBL) is a silent disease caused by a retrovirus [bovine leukaemia virus (BLV)]. BLV is classified into almost 10 genotypes that are distributed in several countries. The present research aimed to describe two BLV gp51 env sequences of strains detected in the state of São Paulo, Brazil and perform a phylogenetic analysis to compare them to other BLV gp51 env sequences of strains around the world. Two bovines from different herds were admitted to the Bovine and Small Ruminant Hospital, School of Veterinary Medicine and Animal Science, University of São Paulo, Brazil. In both, lymphosarcoma was detected and the presence of BLV was confirmed by nested PCR. The neighbour-joining algorithm distance method was used to genotype the BLV sequences by phylogenetic reconstruction, and the maximum likelihood method was used for the phylogenetic reconstruction. The phylogeny estimates were calculated by performing 1000 bootstrap replicates. Analysis of the partial envelope glycoprotein (env) gene sequences from two isolates (25 and 31) revealed two different genotypes of BLV. Isolate 25 clustered with ten genotype 6 isolates from Brazil, Argentina, Thailand and Paraguay. On the other hand, isolate 31 clustered with two genotype 5 isolates (one was also from São Paulo and one was from Costa Rica). The detected genotypes corroborate the results of previous studies conducted in the state of São Paulo, Brazil. The prediction of amino acids showed substitutions, particularly between positions 136 and 150 in 11 out of 13 sequences analysed, including sequences from GenBank. BLV is still important in Brazil and this research should be continued.

  14. Adenovirus, calicivirus and astrovirus detection in fecal samples of hospitalized children with acute gastroenteritis from Campo Grande, MS, Brazil

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    Marcia Sueli Assis Andreasi

    2008-11-01

    Full Text Available We analyzed fecal samples from hospitalized children up to three years of age with acute gastroenteritis at Campo Grande, Mato Grosso do Sul, Brazil, from May 2000-January 2004. Astrovirus and calicivirus were detected by Reverse Transcription-Polymerase Chain Reaction and adenovirus was detected using the Rotavirus and Adenovirus combined immunoenzyme assay. Astrovirus, adenovirus and calicivirus were detected at rates of 3.1%, 3.6% and 7.6%, respectively. These results re-emphasize the need for the establishment of regional vigilance systems to evaluate the impact of enteric viruses on viral gastroenteritis.

  15. Detection of Zika virus in saliva.

    Science.gov (United States)

    Musso, Didier; Roche, Claudine; Nhan, Tu-Xuan; Robin, Emilie; Teissier, Anita; Cao-Lormeau, Van-Mai

    2015-07-01

    During the largest Zika virus (ZIKV) outbreak ever reported that occurred from October 2013 to March 2014 in French Polynesia, we observed that several patients presenting the symptoms of acute phase Zika fever were tested negative in blood by ZIKV real-time PCR (RT-PCR). As we have previously detected ZIKV RNA in the saliva of a young child, we investigated the use of saliva as an alternative sample for routine ZIKV RNA detection. Over a 6 month period, 1,067 samples collected from 855 patients presenting symptoms of Zika fever (saliva only, blood only or both samples) were tested using a specific ZIKV RT-PCR. A medical questionnaire was available for most of the patients. ZIKV was more frequently detected in saliva compared to blood. For the 182 patients with both samples collected, tests were positive for 35 (19.2%) in saliva while negative in blood and tests were positive for 16 (8.8%) in blood while negative in saliva; the difference in mean days after symptoms onset and the percentage of the main symptoms of Zika fever for patients only positive in saliva or in blood was not significant. The use of saliva sample increased the rate of molecular detection of ZIKV at the acute phase of the disease but did not enlarge the window of detection of ZIKV RNA. Saliva was of particular interest when blood was difficult to collect (children and neonates especially). Copyright © 2015 Elsevier B.V. All rights reserved.

  16. Simultaneous detection of Apple mosaic virus in cultivated hazelnuts ...

    African Journals Online (AJOL)

    The most economically damaging ilarvirus affecting hazelnut on a worldwide scale is the related apple mosaic virus (ApMV). Attempts were made to isolate the virus RNA from hazelnut tissues using different extraction methods. The most suitable extraction method that could detect the virus occurring naturally in hazelnut by ...

  17. Zika virus epidemic in Brazil. I. Fatal disease in adults: Clinical and laboratorial aspects.

    Science.gov (United States)

    Azevedo, Raimunda S S; Araujo, Marialva T; Martins Filho, Arnaldo J; Oliveira, Consuelo S; Nunes, Bruno T D; Cruz, Ana C R; Nascimento, Ana G P A C; Medeiros, Rita C; Caldas, Cezar A M; Araujo, Fernando C; Quaresma, Juarez A S; Vasconcelos, Barbara C B; Queiroz, Maria G L; da Rosa, Elizabeth S Travassos; Henriques, Daniele F; Silva, Eliana V P; Chiang, Jannifer O; Martins, Lívia C; Medeiros, Daniele B A; Lima, Juliana A; Nunes, Márcio R T; Cardoso, Jedson F; Silva, Sandro P; Shi, Pei-Yong; Tesh, Robert B; Rodrigues, Sueli G; Vasconcelos, Pedro F C

    2016-12-01

    Zika virus (ZIKV) was first detected in Brazil in May 2015 and the country experienced an explosive epidemic. However, recent studies indicate that the introduction of ZIKV occurred in late 2013. Cases of microcephaly and deaths associated with ZIKV infection were identified in Brazil in November, 2015. To determine the etiology of three fatal adult cases. Here we report three fatal adult cases of ZIKV disease. ZIKV infection in these patients was confirmed by cells culture and/or real-time reverse transcriptase polymerase chain reaction (RT-qPCR) and by antigen detection using immunohistochemical assay. Samples of brain and other selected organs taken at autopsy from three patients were also analyzed by histopathological and immunohistological examination. The first patient, a 36-year-old man with lupus and receiving prednisone therapy, developed a fulminant ZIKV infection. At autopsy, RT-qPCR of blood and tissues was positive for ZIKV RNA, and the virus was cultured from an organ homogenate. The second patient, a previously healthy female, 16 years of age, presented classic symptoms of Zika fever, but later developed severe thrombocytopenia, anemia and hemorrhagic manifestations and died. A blood sample taken on the seventh day of her illness was positive RT-PCR for ZIKV RNA and research in the serum was positive for antinuclear factor fine speckled (1/640), suggesting Evans syndrome (hemolytic anemia an autoimmune disorder with immune thrombocytopenic purpura) secondary to ZIKV infection. The third patient was a 20-year-old woman hospitalized with fever, pneumonia and hemorrhages, who died on 13days after admission. Histopathological changes were observed in all viscera examined. ZIKV antigens were detected by immunohistochemistry in viscera specimens of patients 1 and 3. These three cases demonstrate other potential complications of ZIKV infection, in addition to microcephaly and Guillain-Barre syndrome (GBS), and they suggest that individuals with immune

  18. Detection of caliciviruses associated with acute infantile gastroenteritis in Salvador, an urban center in Northeast Brazil

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    M.P.T.P. Xavier

    2009-05-01

    Full Text Available Acute gastroenteritis caused by viruses is one of the leading causes of infantile morbidity. The aim of the present study was to investigate the presence of human caliciviruses of the genera norovirus and sapovirus in children up to 3 years of age with acute gastroenteritis from low-income communities in the city of Salvador, Brazil. This study is an extension of previous work carried out to establish the profile of the most prevalent enteric pathogens present in these communities. In this report, 139 fecal samples, collected from July 2001 to January 2002 were analyzed by RT-PCR and 13 (9% were positive for human caliciviruses. By sequencing, seven isolates were characterized as norovirus genogroup GII and one as sapovirus genotype GII/1. Sequencing of the previously detected group-A rotaviruses and human astroviruses was also performed and revealed the circulation of rotavirus group A genotypes G1P[8] and G9P[8], and human astrovirus genotypes 6, 7, and 8. No mixed infection was observed. Community-based studies provide geographically representative information on disease burden. However, there are only a few reports in developing countries concerning the genotypes of the most important gastroenteric viruses detected in such communities. The present findings demonstrate the wide diversity of genotypes of the most important viruses responsible for acute gastroenteritis circulating in low-income communities.

  19. Detection of influenza C virus but not influenza D virus in Scottish respiratory samples

    Science.gov (United States)

    Smith, Donald B.; Gaunt, Eleanor R.; Digard, Paul; Templeton, Kate; Simmonds, Peter

    2016-01-01

    Background A newly proposed genus of influenza virus (influenza D) is associated with respiratory disease in pigs and cattle. The novel virus is most closely related to human influenza C virus and can infect ferrets but infection has not been reported in humans. Objectives To ascertain if influenza D virus can be detected retrospectively in patient respiratory samples. Study design 3300 human respiratory samples from Edinburgh, Scotland, covering the period 2006–2008, were screened in pools of 10 by RT-PCR using primers capable of detecting both influenza C and D viruses. Results Influenza D was not detected in any sample. Influenza C was present in 6 samples (0.2%), compared with frequencies of 3.3% and 0.9% for influenza A and B viruses from RT-PCR testing of respiratory samples over the same period. Influenza C virus was detected in samples from individuals 45 years old, with cases occurring throughout the year. Phylogenetic analysis of nearly complete sequences of all seven segments revealed the presence of multiple, reassortant lineages. Conclusion We were unable to detect viruses related to influenza D virus in human respiratory samples. Influenza C virus was less prevalent than influenza A and B viruses, was associated with mild disease in the young (45 years) and comprised multiple, reassortant lineages. Inclusion of influenza C virus as part of a diagnostic testing panel for respiratory infections would be of limited additional value. PMID:26655269

  20. Hepatitis B virus infection profile in hemodialysis patients in Central Brazil: prevalence, risk factors, and genotypes

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    Renata C Ferreira

    2006-09-01

    Full Text Available Hemodialysis patients are at high risk for hepatitis B virus (HBV infection. A survey was conducted in the hemodialysis population of the state of Goiás, Central Brazil, aiming to assess the prevalence of HBV infection, to analyse associated risk factors, and also to investigate HBV genotypes distribution. A total of 1095 patients were interviewed in 15 dialysis units. Serum samples were screened for HBV serological markers by enzyme-linked immunosorbent assay. Hepatitis B surface antigen (HBsAg positive samples were tested for HBV DNA by polymerase chain reaction and genotyped by restriction fragment length polymorphism. Global HBV infection prevalence was 29.8% (95% CI: 27.1-32.5. Multivariate analysis of risk factors showed that male gender, length of time on hemodialysis, and blood transfusion before 1993 were associated with HBV positivity. HBV DNA was detected in 65.4% (17/26 of the HBsAg-positive samples. Thirteen of 17 HBV DNA positive samples were genotyped. Genotype D (61.5% was predominant, followed by A (30.8%, while genotype F was detected in only one (7.7% sample.

  1. Zika Virus Outbreak in Rio de Janeiro, Brazil: Clinical Characterization, Epidemiological and Virological Aspects.

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    Patrícia Brasil

    2016-04-01

    Full Text Available In 2015, Brazil was faced with the cocirculation of three arboviruses of major public health importance. The emergence of Zika virus (ZIKV presents new challenges to both clinicians and public health authorities. Overlapping clinical features between diseases caused by ZIKV, Dengue (DENV and Chikungunya (CHIKV and the lack of validated serological assays for ZIKV make accurate diagnosis difficult.The outpatient service for acute febrile illnesses in Fiocruz initiated a syndromic clinical observational study in 2007 to capture unusual presentations of DENV infections. In January 2015, an increase of cases with exanthematic disease was observed. Trained physicians evaluated the patients using a detailed case report form that included clinical assessment and laboratory investigations. The laboratory diagnostic algorithm included assays for detection of ZIKV, CHIKV and DENV. 364 suspected cases of Zika virus disease were identified based on clinical criteria between January and July 2015. Of these, 262 (71.9% were tested and 119 (45.4% were confirmed by the detection of ZIKV RNA. All of the samples with sequence information available clustered within the Asian genotype.This is the first report of a ZIKV outbreak in the state of Rio de Janeiro, based on a large number of suspected (n = 364 and laboratory confirmed cases (n = 119. We were able to demonstrate that ZIKV was circulating in Rio de Janeiro as early as January 2015. The peak of the outbreak was documented in May/June 2015. More than half of the patients reported headache, arthralgia, myalgia, non-purulent conjunctivitis, and lower back pain, consistent with the case definition of suspected ZIKV disease issued by the Pan American Health Organization (PAHO. However, fever, when present, was low-intensity and short-termed. In our opinion, pruritus, the second most common clinical sign presented by the confirmed cases, should be added to the PAHO case definition, while fever could be given less

  2. Zika Virus Outbreak in Rio de Janeiro, Brazil: Clinical Characterization, Epidemiological and Virological Aspects.

    Science.gov (United States)

    Brasil, Patrícia; Calvet, Guilherme Amaral; Siqueira, André Machado; Wakimoto, Mayumi; de Sequeira, Patrícia Carvalho; Nobre, Aline; Quintana, Marcel de Souza Borges; Mendonça, Marco Cesar Lima de; Lupi, Otilia; de Souza, Rogerio Valls; Romero, Carolina; Zogbi, Heruza; Bressan, Clarisse da Silveira; Alves, Simone Sampaio; Lourenço-de-Oliveira, Ricardo; Nogueira, Rita Maria Ribeiro; Carvalho, Marilia Sá; de Filippis, Ana Maria Bispo; Jaenisch, Thomas

    2016-04-01

    In 2015, Brazil was faced with the cocirculation of three arboviruses of major public health importance. The emergence of Zika virus (ZIKV) presents new challenges to both clinicians and public health authorities. Overlapping clinical features between diseases caused by ZIKV, Dengue (DENV) and Chikungunya (CHIKV) and the lack of validated serological assays for ZIKV make accurate diagnosis difficult. The outpatient service for acute febrile illnesses in Fiocruz initiated a syndromic clinical observational study in 2007 to capture unusual presentations of DENV infections. In January 2015, an increase of cases with exanthematic disease was observed. Trained physicians evaluated the patients using a detailed case report form that included clinical assessment and laboratory investigations. The laboratory diagnostic algorithm included assays for detection of ZIKV, CHIKV and DENV. 364 suspected cases of Zika virus disease were identified based on clinical criteria between January and July 2015. Of these, 262 (71.9%) were tested and 119 (45.4%) were confirmed by the detection of ZIKV RNA. All of the samples with sequence information available clustered within the Asian genotype. This is the first report of a ZIKV outbreak in the state of Rio de Janeiro, based on a large number of suspected (n = 364) and laboratory confirmed cases (n = 119). We were able to demonstrate that ZIKV was circulating in Rio de Janeiro as early as January 2015. The peak of the outbreak was documented in May/June 2015. More than half of the patients reported headache, arthralgia, myalgia, non-purulent conjunctivitis, and lower back pain, consistent with the case definition of suspected ZIKV disease issued by the Pan American Health Organization (PAHO). However, fever, when present, was low-intensity and short-termed. In our opinion, pruritus, the second most common clinical sign presented by the confirmed cases, should be added to the PAHO case definition, while fever could be given less emphasis

  3. VirusDetect: An automated pipeline for efficient virus discovery using deep sequencing of small RNAs

    Science.gov (United States)

    Accurate detection of viruses in plants and animals is critical for agriculture production and human health. Deep sequencing and assembly of virus-derived siRNAs has proven to be a highly efficient approach for virus discovery. However, to date no computational tools specifically designed for both k...

  4. Practical and sensitive screening strategy for detection of influenza virus.

    Science.gov (United States)

    Newton, Duane W; Mellen, Cindy F; Baxter, Barbara D; Atmar, Robert L; Menegus, Marilyn A

    2002-11-01

    This study evaluated the performance of Directigen FluA combined with a 3-day flu screening culture for the detection of influenza virus. This abbreviated protocol was a useful and effective tool and resulted in a substantial reduction in time, effort, and money spent, while not compromising sensitivity of influenza virus detection.

  5. Molecular identification based on coat protein sequences of the Barley yellow dwarf virus from Brazil

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    Talita Bernardon Mar

    2013-12-01

    Full Text Available Yellow dwarf disease, one of the most important diseases of cereal crops worldwide, is caused by virus species belonging to the Luteoviridae family. Forty-two virus isolates obtained from oat (Avena sativa L., wheat (Triticum aestivum L., barley (Hordeum vulgare L., corn (Zea mays L., and ryegrass (Lolium multiflorum Lam. collected between 2007 and 2008 from winter cereal crop regions in southern Brazil were screened by polymerase chain reaction (PCR with primers designed on ORF 3 (coat protein - CP for the presence of Barley yellow dwarf virus and Cereal yellow dwarf virus (B/CYDV. PCR products of expected size (~357 bp for subgroup II and (~831 bp for subgroup I were obtained for three and 39 samples, respectively. These products were cloned and sequenced. The subgroup II 3' partial CP amino acid deduced sequences were identified as BYDV-RMV (92 - 93 % of identity with "Illinois" Z14123 isolate. The complete CP amino acid deduced sequences of subgroup I isolates were confirmed as BYDV-PAV (94 - 99 % of identity and established a very homogeneous group (identity higher than 99 %. These results support the prevalence of BYDV-PAV in southern Brazil as previously diagnosed by Enzyme-Linked Immunosorbent Assay (ELISA and suggest that this population is very homogeneous. To our knowledge, this is the first report of BYDV-RMV in Brazil and the first genetic diversity study on B/CYDV in South America.

  6. Molecular Characterization of Subtype H11N9 Avian Influenza Virus Isolated from Shorebirds in Brazil.

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    Renata Hurtado

    Full Text Available Migratory aquatic birds play an important role in the maintenance and spread of avian influenza viruses (AIV. Many species of aquatic migratory birds tend to use similar migration routes, also known as flyways, which serve as important circuits for the dissemination of AIV. In recent years there has been extensive surveillance of the virus in aquatic birds in the Northern Hemisphere; however in contrast only a few studies have been attempted to detect AIV in wild birds in South America. There are major flyways connecting South America to Central and North America, whereas avian migration routes between South America and the remaining continents are uncommon. As a result, it has been hypothesized that South American AIV strains would be most closely related to the strains from North America than to those from other regions in the world. We characterized the full genome of three AIV subtype H11N9 isolates obtained from ruddy turnstones (Arenaria interpres on the Amazon coast of Brazil. For all gene segments, all three strains consistently clustered together within evolutionary lineages of AIV that had been previously described from aquatic birds in North America. In particular, the H11N9 isolates were remarkably closely related to AIV strains from shorebirds sampled at the Delaware Bay region, on the Northeastern coast of the USA, more than 5000 km away from where the isolates were retrieved. Additionally, there was also evidence of genetic similarity to AIV strains from ducks and teals from interior USA and Canada. These findings corroborate that migratory flyways of aquatic birds play an important role in determining the genetic structure of AIV in the Western hemisphere, with a strong epidemiological connectivity between North and South America.

  7. Neuraminidase as an enzymatic marker for detecting airborne Influenza virus and other viruses.

    Science.gov (United States)

    Turgeon, Nathalie; Toulouse, Marie-Josée; Ho, Jim; Li, Dongqing; Duchaine, Caroline

    2017-02-01

    Little information is available regarding the effectiveness of air samplers to collect viruses and regarding the effects of sampling processes on viral integrity. The neuraminidase enzyme is present on the surface of viruses that are of agricultural and medical importance. It has been demonstrated that viruses carrying this enzyme can be detected using commercial substrates without having to process the sample by methods such as RNA extraction. This project aims at evaluating the effects of 3 aerosol-sampling devices on the neuraminidase enzyme activity of airborne viruses. The purified neuraminidase enzymes from Clostridium perfringens, a strain of Influenza A (H1N1) virus, the FluMist influenza vaccine, and the Newcastle disease virus were used as models. The neuraminidase models were aerosolized in aerosol chambers and sampled with 3 different air samplers (SKC BioSampler, 3-piece cassettes with polycarbonate filters, and Coriolis μ) to assess the effect on neuraminidase enzyme activity. Our results demonstrated that Influenza virus and Newcastle disease virus neuraminidase enzymes are resistant to aerosolization and sampling with all air samplers tested. Moreover, we demonstrated that the enzymatic neuraminidase assay is as sensitive as RT-qPCR for detecting low concentrations of Influenza virus and Newcastle disease virus. Therefore, given the sensitivity of the assay and its compatibility with air sampling methods, viruses carrying the neuraminidase enzyme can be rapidly detected from air samples using neuraminidase activity assay without having to preprocess the samples.

  8. Design of microarray probes for virus identification and detection of emerging viruses at the genus level

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    Ho Mei-Shang

    2006-04-01

    Full Text Available Abstract Background Most virus detection methods are geared towards the detection of specific single viruses or just a few known targets, and lack the capability to uncover the novel viruses that cause emerging viral infections. To address this issue, we developed a computational method that identifies the conserved viral sequences at the genus level for all viral genomes available in GenBank, and established a virus probe library. The virus probes are used not only to identify known viruses but also for discerning the genera of emerging or uncharacterized ones. Results Using the microarray approach, the identity of the virus in a test sample is determined by the signals of both genus and species-specific probes. The genera of emerging and uncharacterized viruses are determined based on hybridization of the viral sequences to the conserved probes for the existing viral genera. A detection and classification procedure to determine the identity of a virus directly from detection signals results in the rapid identification of the virus. Conclusion We have demonstrated the validity and feasibility of the above strategy with a small number of viral samples. The probe design algorithm can be applied to any publicly available viral sequence database. The strategy of using separate genus and species probe sets enables the use of a straightforward virus identity calculation directly based on the hybridization signals. Our virus identification strategy has great potential in the diagnosis of viral infections. The virus genus and specific probe database and the associated summary tables are available at http://genestamp.sinica.edu.tw/virus/index.htm.

  9. Design of microarray probes for virus identification and detection of emerging viruses at the genus level.

    Science.gov (United States)

    Chou, Cheng-Chung; Lee, Te-Tsui; Chen, Chun-Houh; Hsiao, Hsiang-Yun; Lin, Yi-Ling; Ho, Mei-Shang; Yang, Pan-Chyr; Peck, Konan

    2006-04-28

    Most virus detection methods are geared towards the detection of specific single viruses or just a few known targets, and lack the capability to uncover the novel viruses that cause emerging viral infections. To address this issue, we developed a computational method that identifies the conserved viral sequences at the genus level for all viral genomes available in GenBank, and established a virus probe library. The virus probes are used not only to identify known viruses but also for discerning the genera of emerging or uncharacterized ones. Using the microarray approach, the identity of the virus in a test sample is determined by the signals of both genus and species-specific probes. The genera of emerging and uncharacterized viruses are determined based on hybridization of the viral sequences to the conserved probes for the existing viral genera. A detection and classification procedure to determine the identity of a virus directly from detection signals results in the rapid identification of the virus. We have demonstrated the validity and feasibility of the above strategy with a small number of viral samples. The probe design algorithm can be applied to any publicly available viral sequence database. The strategy of using separate genus and species probe sets enables the use of a straightforward virus identity calculation directly based on the hybridization signals. Our virus identification strategy has great potential in the diagnosis of viral infections. The virus genus and specific probe database and the associated summary tables are available at http://genestamp.sinica.edu.tw/virus/index.htm.

  10. Nucleic acid hybridization techniques for the detection of bluetongue virus

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    Schoepp, R.J.

    1989-01-01

    Virus isolation, antigen detection, and in situ hybridization were compared in their abilities to detect in cell culture, the five serotypes of bluetongue virus (BTV) occurring in the United States, serotypes 2, 10, 11, 13, and 17. For isolation, virus was propagated in baby hamster kidney (BHK-21) cell culture. For antigen detection, two techniques, indirect fluorescent-antibody (IFA) and enzyme immunocytoassay (EICA) were used. For in situ hybridization, a complementary DNA (cDNA) of the L3 RNA genome segment of BTV, serotype 17 (BTV-17) labeled with {sup 35}S was used as a group-specific probe. Virus isolation was the most sensitive technique, often detecting input virus and then detecting virus throughout the course of the study. IFA and EICA were of similar sensitivity and detected BTV antigen shortly after detection of virus by isolation. A direct-blot hybridization technique using a {sup 32}P-labeled, strand-specific RNA transcript probe was developed, optimized, and used to detect BTV in pools of infected Culicoides variipennis midges. The technique was able to detect as few as one infected Culicoides midge in a pool of 100 and as little as 3.5 log{sub 10} TCID{sub 50} per ml of virus. A sandwich hybridization technique was developed and used to detect BTV in pools of infected Culicoides variipennis midges. The sandwich hybridization technique used a single-stranded DNA catcher sequence bound to a solid support and a {sup 32}P-labeled, single-stranded RNA detector sequence. Sandwich hybridization was compared to direct blot hybridization using a strand-specific RNA transcript probe or a cDNA probe. Sandwich hybridization was able to detect as few as one infected Culicoides midge in a pool of 50; however, the technique was approximately tenfold less sensitive than direct blot hybridization.

  11. Hepatitis C and hepatitis B virus infection in different hemodialysis units in Belo Horizonte, Minas Gerais, Brazil

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    Busek Solange U

    2002-01-01

    Full Text Available The prevalence, virological and epidemilogical aspects of the hepatitis C virus (HCV and the hepatitis B virus (HBV infections vary among hemodialysis patients in different countries. Aiming at analyzing these aspects of HCV and HBV infections in hemodialysis patients in Belo Horizonte, MG, Brazil, we studied three hemodialysis units including 434 patients. Serology was used to detect anti-HCV and HBsAg. Reverse trancriptase nested polymerase chain reaction (RT-nested-PCR of the 5'-noncoding region was used to detect circulating HCV RNA and restriction fragment length polymorphism analysis for genotyping. Seroprevalence varied from 26.5% to 11.1% for hepatitis C and from 5.9% to 0% for hepatitis B. Risk factors observed for HBV and/or HCV infections were the number of patients per dialysis unit, duration of treatment, number of clinics attended, number of blood units transfused, and lower level scholarity. Alanine aminotransferase levels were altered with a higher frequency in HBV or HCV seropositive patients. Half of ten patients, negative for anti-HCV, had detectable viremia by RT-nested-PCR, indicating that this technique should be used to confirm infections in this group of patients. The HCV genotype 1 was the most frequently observed, followed by the genotype 2, but no correlation was detected between genotype and clinical or epidemiological data.

  12. Phylogeography and evolutionary history of hepatitis B virus genotype F in Brazil.

    Science.gov (United States)

    Mello, Francisco C A; Araujo, Oscar C; Lago, Barbara V; Motta-Castro, Ana Rita C; Moraes, Marcia Terezinha B; Gomes, Selma A; Bello, Gonzalo; Araujo, Natalia M

    2013-07-16

    Hepatitis B virus (HBV) genotype F (HBV/F) is considered to be indigenous to the Americas, but its emergence and spread in the continent remain unknown. Previously, only two HBV/F complete genome sequences from Brazil were available, limiting the contribution of Brazilian isolates to the phylogenetic studies of HBV/F. The present study was carried out to assess the proportion and geographic distributions of HBV/F subgenotypes in Brazil, to determine the full-length genomic sequences of HBV/F isolates from different Brazilian geographic regions, and to investigate the detailed evolutionary history and phylogeography of HBV/F in Brazil. Complete HBV/F genomes isolated from 12 Brazilian patients, representing the HBV/F subgenotypes circulating in Brazil, were sequenced and analyzed together with sequences retrieved from GenBank, using the Bayesian coalescent and phylogeographic framework. Phylogenetic analysis using all Brazilian HBV/F S-gene sequences available in GenBank showed that HBV/F2a is found at higher frequencies countrywide and corresponds to all sequences isolated in the Brazilian Amazon Basin. In addition, the evolutionary analysis using complete genome sequences estimated an older median ancestral age for the Brazilian HBV/F2a compared to the Brazilian HBV/F1b and HBV/F4 subgenotypes, suggesting that HBV/F2a represents the original native HBV of Brazil. The phylogeographic patterns suggested a north-to-south flow of HBV/F2a from Venezuela to Brazil, whereas HBV/F1b and HBV/F4 strains appeared to have spread from Argentina to Brazil. This study suggests a plausible route of introduction of HBV/F subgenotypes in Brazil and demonstrates the usefulness of recently developed computational tools for investigating the evolutionary history of HBV.

  13. Oral Susceptibility to Yellow Fever Virus of Aedes aegypti from Brazil

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    Lourenço-de-Oliveira Ricardo

    2002-01-01

    Full Text Available The oral susceptibility to yellow fever virus was evaluated in 23 Aedes aegypti samples from Brazil. Six Ae. aegypti samples from Africa, America and Asia were also tested for comparison. Mosquito samples from Asia showed the highest infection rates. Infection rates for the Brazilian Ae. aegypti reached 48.6%, but were under 13% in 60% of sample tested. We concluded that although the low infection rates estimated for some Brazilian mosquito samples may not favor the establishment of urban cycle of yellow fever in some parts of the country, the founding of Ae. aegypti of noteworthy susceptibility to the virus in cities located in endemic and transition areas of sylvatic yellow fever, do pose a threat of the re-emergence of the urban transmission of the disease in Brazil.

  14. Detection and diagnosis of rice-infecting viruses

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    Tamaki Uehara Ichiki

    2013-10-01

    Full Text Available Rice-infecting viruses have caused serious damage to rice production in Asian, American, and African countries, where about 30 rice viruses and diseases have been reported. To control these diseases, developing accurate, quick methods to detect and diagnose the viruses in the host plants and any insect vectors of the viruses is very important. Based on an antigen–antibody reaction, serological methods such as latex agglutination reaction (LAR and enzyme-linked immunosorbent assay (ELISA have advanced to detect viral particles or major proteins derived from viruses. They aid in forecasting disease and surveying disease spread and are widely used for virus detection at plant protection stations and research laboratories. From the early 2000s, based on sequence information for the target virus, several other methods such as reverse transcription-polymerase chain reaction (RT-PCR and reverse transcription- loop-mediated isothermal amplification (RT-LAMP have been developed that are sensitive, rapid, and able to differentiate closely related viruses. Recent techniques such as real-time RT-PCR can be used to quantify the pathogen in target samples and monitor population dynamics of a virus, and metagenomic analyses using next-generation sequencing and microarrays show potential for use in the diagnosis of rice diseases.

  15. A multiplex PCR for detection of six viruses in ducks.

    Science.gov (United States)

    Wang, Yongjuan; Zhu, Shanyuan; Hong, Weiming; Wang, Anping; Zuo, Weiyong

    2017-10-01

    In this study, six pairs of specific primers that can amplify DNA fragments of different sizes were designed and synthesized according to viral protein gene sequences published in GenBank. Then, a multiplex PCR method was established for rapid detection of duck hepatitis virus 1, duck plague virus, duck Tembusu virus, muscovy duck parvovirus, muscovy duck reovirus, and duck H9N2 avian influenza virus, and achieve simple and rapid detection of viral diseases in ducks. Single PCR was used to confirm primer specificity, and PCR conditions were optimized to construct a multiplex PCR system. Specificity and sensitivity assays were also developed. The multiplex PCR was used to detect duck embryos infected with mixed viruses and those with clinically suspected diseases to verify the feasibility of the multiplex PCR. Results show that the primers can specifically amplify target fragments, without any cross-amplification with other viruses. The multiplex PCR system can amplify six DNA fragments from the pooled viral genomes and specifically detect nucleic acids of the six duck susceptible viruses when the template amount is 10 2 copies/μl. In addition, the system can be used to detect viral nucleic acids in duck embryos infected with the six common viruses. The detection results for clinical samples are consistent with those detected by single PCR. Therefore, the established multiplex PCR method can perform specific, sensitive, and high-throughput detection of six duck-infecting viruses and can be applied to clinical identification and diagnosis of viral infection in ducks. Copyright © 2017. Published by Elsevier B.V.

  16. Microcephaly Case Fatality Rate Associated with Zika Virus Infection in Brazil: Current Estimates.

    Science.gov (United States)

    Cunha, Antonio José Ledo Alves da; de Magalhães-Barbosa, Maria Clara; Lima-Setta, Fernanda; Medronho, Roberto de Andrade; Prata-Barbosa, Arnaldo

    2017-05-01

    Considering the currently confirmed cases of microcephaly and related deaths associated with Zika virus in Brazil, the estimated case fatality rate is 8.3% (95% confidence interval: 7.2-9.6). However, a third of the reported cases remain under investigation. If the confirmation rates of cases and deaths are the same in the future, the estimated case fatality rate will be as high as 10.5% (95% confidence interval: 9.5-11.7).

  17. Microcephaly and Zika virus: a clinical and epidemiological analysis of the current outbreak in Brazil,

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    Magda Lahorgue Nunes

    2016-06-01

    Full Text Available Abstract Objective: This study aimed to critically review the literature available regarding the Zika virus outbreak in Brazil and its possible association with microcephaly cases. Sources: Experts from Instituto do Cérebro do Rio Grande do Sul performed a critical (nonsystematic literature review regarding different aspects of the Zika virus outbreak in Brazil, such as transmission, epidemiology, diagnostic criteria, and its possible association with the increase of microcephaly reports. The PubMed search using the key word “Zika virus” in February 2016 yielded 151 articles. The manuscripts were reviewed, as well as all publications/guidelines from the Brazilian Ministry of Health, World Health Organization and Centers for Disease Control and Prevention (CDC – United States. Summary of findings: Epidemiological data suggest a temporal association between the increased number of microcephaly notifications in Brazil and outbreak of Zika virus, primarily in the Brazil's Northeast. It has been previously documented that many different viruses might cause congenital acquired microcephaly. Still there is no consensus on the best curve to measure cephalic circumference, specifically in preterm neonates. Conflicting opinions regarding the diagnosis of microcephaly (below 2 or 3 standard deviations that should be used for the notifications were also found in the literature. Conclusion: The development of diagnostic techniques that confirm a cause–effect association and studies regarding the physiopathology of the central nervous system impairment should be prioritized. It is also necessary to strictly define the criteria for the diagnosis of microcephaly to identify cases that should undergo an etiological investigation.

  18. GB virus C/hepatitis G virus infection in dialysis patients and kidney transplant recipients in central Brazil

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    Ramon Ramos Filho

    2004-10-01

    Full Text Available In order to investigate the prevalence of GB virus C (GBV-C/hepatitis G virus (HGV infection in dialysis patients and kidney transplant recipients in Central Brazil and also to analyze the virus genotypes distribution, a total of 123 patients including 98 on hemodialysis, 13 on continuous ambulatory peritoneal dialysis treatment, and 12 who received kidney transplantation were interviewed in one unit of dialysis treatment in Goiânia city. Blood samples were collected and serum samples tested for GBV-C/HGV RNA by polymerase chain reaction. Genotypes were determined by restriction fragment length polymorphism (RFLP analysis. Eighteen samples were GBV-C/HGV RNA-positive, resulting in an overall prevalence of 14.6% (95% CI: 9.2-21.7. A high positivity for GBV-C/HGV RNA was observed in patients who had received kidney transplant (16.7%, followed by those on hemodialysis (15.3%, and peritoneal dialysis (7.7%. RFLP analysis revealed the presence of genotypes 1, 2, and 3 of GBV-C/HGV; more precisely, 9 (50% samples were found belonging to the 2b subtype, 4 (22% to the 2a subtype, 3 (17% to genotype 1, and 2 (11% to genotype 3. The present data indicate an intermediate prevalence of GBV-C/HGV infection among dialysis patients and kidney transplant recipients in Central Brazil. Genotype 2 (subtype 2b seems to be the most prevalent GBV-C/HGV genotype in our region.

  19. Detection of bla KPC-2 in Proteus mirabilis in Brazil

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    Adriane Borges Cabral

    2015-02-01

    Full Text Available INTRODUCTION : Infections caused by Klebsiella pneumoniae carbapenemase (KPC-producing isolates pose a major worldwide public health problem today. METHODS : A carbapenem-resistant Proteus mirabilis clinical isolate was investigated for plasmid profiles and the occurrence of β-lactamase genes. RESULTS : The isolate exhibited resistance to ertapenem and imipenem and was susceptible to meropenem, polymyxin, and tigecycline. Five plasmids were identified in this isolate. DNA sequencing analysis revealed the presence of bla KPC-2 and bla TEM-1 genes. An additional PCR using plasmid DNA confirmed that bla KPC-2 was present in one of these plasmids. Conclusions: We report the detection of bla KPC-2 in P. mirabilis in Brazil for the first time. This finding highlights the continuous transfer of bla KPC between bacterial genera, which presents a serious challenge to the prevention of infection by multidrug-resistant bacteria.

  20. Prevalence and genotypes of GB Virus C/Hepatitis G virus among blood donors in Central Brazil

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    Luciana A Oliveira

    2002-10-01

    Full Text Available A survey was conducted in a blood donor population of Central Brazil aiming to investigate the prevalence of GB virus C (GBV-C/hepatitis G virus (HGV infection and also to analyze the virus genotypes distribution. A total of 241 voluntary blood donors were interviewed at the State Blood Bank in Goiânia, State of Goiás, Brazil. Blood samples were collected and serum samples tested for GBV-C/HGV RNA by polymerase chain reaction. Genotypes were determined by restriction fragment length polymorphism (RFLP analysis. Seventeen samples were GBV-C/HGV RNA-positive, resulting in a prevalence of 7.1% (95% CI: 4.2-11.1. A significant trend of GBV-C/HGV RNA positivity in relation to age was observed, with the highest prevalence in donors between 29-39 years old. Ten infected individuals were characterized by reporting parenteral (30%, sexual (18%, both (6% and intrafamiliar (6% transmission. However, 7 (40% GBV-C/HGV RNA-positive donors did not mention any potential transmission route. RFLP analysis revealed the presence of genotypes 1 and 2 of GBV-C/HGV; more precisely, 10 (58.9% samples were found belonging to the 2b subtype, 4 (23.5% to the 2a subtype, and 3 (17.6% to genotype 1. The present data indicate an intermediate endemicity of GBV-C/HGV infection among this blood donor population, and a predominant circulation of genotype 2 (subtype 2b in Central Brazil.

  1. Detection of pathogenic viruses in sewage provided early warnings of hepatitis A virus and norovirus outbreaks.

    Science.gov (United States)

    Hellmér, Maria; Paxéus, Nicklas; Magnius, Lars; Enache, Lucica; Arnholm, Birgitta; Johansson, Annette; Bergström, Tomas; Norder, Heléne

    2014-11-01

    Most persons infected with enterically transmitted viruses shed large amounts of virus in feces for days or weeks, both before and after onset of symptoms. Therefore, viruses causing gastroenteritis may be detected in wastewater, even if only a few persons are infected. In this study, the presence of eight pathogenic viruses (norovirus, astrovirus, rotavirus, adenovirus, Aichi virus, parechovirus, hepatitis A virus [HAV], and hepatitis E virus) was investigated in sewage to explore whether their identification could be used as an early warning of outbreaks. Samples of the untreated sewage were collected in proportion to flow at Ryaverket, Gothenburg, Sweden. Daily samples collected during every second week between January and May 2013 were pooled and analyzed for detection of viruses by concentration through adsorption to milk proteins and PCR. The largest amount of noroviruses was detected in sewage 2 to 3 weeks before most patients were diagnosed with this infection in Gothenburg. The other viruses were detected at lower levels. HAV was detected between weeks 5 and 13, and partial sequencing of the structural VP1protein identified three different strains. Two strains were involved in an ongoing outbreak in Scandinavia and were also identified in samples from patients with acute hepatitis A in Gothenburg during spring of 2013. The third strain was unique and was not detected in any patient sample. The method used may thus be a tool to detect incipient outbreaks of these viruses and provide early warning before the causative pathogens have been recognized in health care. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  2. Ilheus virus isolation in the Pantanal, west-central Brazil.

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    Pauvolid-Corrêa, Alex; Kenney, Joan L; Couto-Lima, Dinair; Campos, Zilca M S; Schatzmayr, Hermann G; Nogueira, Rita M R; Brault, Aaron C; Komar, Nicholas

    2013-01-01

    The wetlands of the Brazilian Pantanal host large concentrations of diverse wildlife species and hematophagous arthropods, conditions that favor the circulation of zoonotic arboviruses. A recent study from the Nhecolândia sub-region of Pantanal reported serological evidence of various flaviviruses, including West Nile virus and Ilheus virus (ILHV). According to the age of seropositive horses, at least three flaviviruses, including ILHV, circulated in the Brazilian Pantanal between 2005 and 2009. To extend this study, we collected 3,234 adult mosquitoes of 16 species during 2009 and 2010 in the same sub-region. Mosquito pool homogenates were assayed for infectious virus on C6/36 and Vero cell monolayers and also tested for flaviviral RNA by a group-specific real-time RT-PCR. One pool containing 50 non-engorged female specimens of Aedes scapularis tested positive for ILHV by culture and for ILHV RNA by real-time RT-PCR, indicating a minimum infection rate of 2.5 per 1000. Full-length genomic sequence exhibited 95% identity to the only full genome sequence available for ILHV. The present data confirm the circulation of ILHV in the Brazilian Pantanal.

  3. Spirometry filters can be used to detect exhaled respiratory viruses.

    Science.gov (United States)

    Mitchell, Alicia B; Mourad, Bassel; Tovey, Euan; Buddle, Lachlan; Peters, Matthew; Morgan, Lucy; Oliver, Brian G

    2016-09-26

    Respiratory viruses are very common in the community and contribute to the burden of illness for patients with chronic respiratory diseases, including acute exacerbations. Traditional sampling methods are invasive and problematic to repeat. Accordingly, we explored whether respiratory viruses could be isolated from disposable spirometry filters and whether detection of viruses in this context represented presence in the upper or lower respiratory tract. Discovery (n  =  53) and validation (n  =  49) cohorts were recruited from a hospital outpatient department during two different time periods. Spirometry mouthpiece filters were collected from all participants. Respiratory secretions were sampled from the upper and lower respiratory tract by nasal washing (NW), sputum, and bronchoalveolar lavage (BAL). All samples were examined using RT-PCR to identify a panel of respiratory viruses (rhinovirus, respiratory syncytial virus, influenza A, influenza B, parainfluenza virus 1, 2 & 3, and human metapneumovirus). Rhinovirus was quantified using qPCR. Paired filter-NW samples (n  =  29), filter-sputum samples (n  =  24), filter-BAL samples (n  =  39) and filter-NW-BAL samples (n  =  10) provided a range of comparisons. At least one virus was detected in any sample in 85% of participants in the discovery cohort versus 45% in the validation cohort. Overall, 72% of viruses identified in the paired comparator method matched those detected in spirometry filters. There was a high correlation between viruses identified in spirometry filters compared with viruses identified in both the upper and lower respiratory tract using traditional sampling methods. Our results suggest that examination of spirometry filters may be a novel and inexpensive sampling method for the presence of respiratory viruses in exhaled breath.

  4. First detection of bluetongue virus serotype 14 in Poland.

    Science.gov (United States)

    Orłowska, Anna; Trębas, Paweł; Smreczak, Marcin; Marzec, Anna; Żmudziński, Jan F

    2016-07-01

    Here, we present the first detected cases of bluetongue virus (BTV) in native cattle from Poland. The virus was found in animals located near the Polish-Belarusian and Polish-Lithuanian borders. The positive animals were detected through an official epidemiological surveillance program. A combination of type-specific real-time RT-PCR and phylogenetic tests revealed the presence of BTV serotype 14 (BTV-14). This serotype is highly homologous to the vaccine strain and BTV-14 present in Russia, Lithuania, and Spain (from an animal imported from Lithuania). The most probable route of virus introduction to Poland was transmission through midges. All of the cases were subclinical.

  5. Epstein-Barr virus DNA associated with gastric adenocarcinoma and adjacent non-cancerous mucosa in patients from Manaus, Brazil.

    Science.gov (United States)

    de Aquino, P F; Carvalho, P C; da Gama Fischer, J S; de Souza, A Q L; Viana, J S; Chalub, S R S; de Souza, A D L; Carvalho, M G C

    2012-12-17

    Gastric cancer is one of most frequent causes of death in Brazil. The city of Manaus has one of the highest incidences of this disease in Brazil. The Epstein-Barr virus (EBV) is a ubiquitous herpesvirus that is classified as a group 1 carcinogen by the International Agency for Research on Cancer. We obtained biopsies from 6 control subjects and 10 patients with gastric carcinomas living in Manaus. In the patients, the samples were taken from tumors and from adjacent non-cancerous mucosa. These samples were screened for EBV DNA by PCR to amplify the 288-bp fragments from the Bam M region. The EBV DNA was detected in 8 of the 10 tumor cases and in none of the 6 control subjects. In the positively identified samples, EBV DNA was detected in five corresponding resection margins. Previous research indicated only a weak association between EBV and gastric cancer. We suggest that EBV should be considered as a risk factor for gastric adenocarcinomas in Manaus.

  6. On the origin and timing of Zika virus introduction in Brazil.

    Science.gov (United States)

    Massad, E; Burattini, M Nascimento; Khan, K; Struchiner, C J; Coutinho, F A B; Wilder-Smith, A

    2017-08-01

    The timing and origin of Zika virus (ZIKV) introduction in Brazil has been the subject of controversy. Initially, it was assumed that the virus was introduced during the FIFA World Cup in June-July 2014. Then, it was speculated that ZIKV may have been introduced by athletes from French Polynesia (FP) who competed in a canoe race in Rio de Janeiro in August 2014. We attempted to apply mathematical models to determine the most likely time window of ZIKV introduction in Brazil. Given that the timing and origin of ZIKV introduction in Brazil may be a politically sensitive issue, its determination (or the provision of a plausible hypothesis) may help to prevent undeserved blame. We used a simple mathematical model to estimate the force of infection and the corresponding individual probability of being infected with ZIKV in FP. Taking into account the air travel volume from FP to Brazil between October 2013 and March 2014, we estimated the expected number of infected travellers arriving at Brazilian airports during that period. During the period between December 2013 and February 2014, 51 individuals travelled from FP airports to 11 Brazilian cities. Basing on the calculated force of ZIKV infection (the per capita rate of new infections per time unit) and risk of infection (probability of at least one new infection), we estimated that 18 (95% CI 12-22) individuals who arrived in seven of the evaluated cities were infected. When basic ZIKV reproduction numbers greater than one were assumed in the seven evaluated cities, ZIKV could have been introduced in any one of the cities. Based on the force of infection in FP, basic reproduction ZIKV number in selected Brazilian cities, and estimated travel volume, we concluded that ZIKV was most likely introduced and established in Brazil by infected travellers arriving from FP in the period between October 2013 and March 2014, which was prior to the two aforementioned sporting events.

  7. Neutralising antibodies for Mayaro virus in Pantanal, Brazil

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    Alex Pauvolid-Corrêa

    2015-02-01

    Full Text Available The Pantanal hosts diverse wildlife species and therefore is a hotspot for arbovirus studies in South America. A serosurvey for Mayaro virus (MAYV, eastern (EEEV, western (WEEV and Venezuelan (VEEV equine encephalitis viruses was conducted with 237 sheep, 87 free-ranging caimans and 748 equids, including 37 collected from a ranch where a neurologic disorder outbreak had been recently reported. Sera were tested for specific viral antibodies using plaque-reduction neutralisation test. From a total of 748 equids, of which 264 were immunised with vaccine composed of EEEV and WEEV and 484 had no history of immunisation, 10 (1.3% were seropositive for MAYV and two (0.3% for VEEV using criteria of a ≥ 4-fold antibody titre difference. Among the 484 equids without history of immunisation, 48 (9.9% were seropositive for EEEV and four (0.8% for WEEV using the same criteria. Among the sheep, five were sero- positive for equine encephalitis alphaviruses, with one (0.4% for EEEV, one (0.4% for WEEV and three (1.3% for VEEV. Regarding free-ranging caimans, one (1.1% and three (3.4%, respectively, had low titres for neutralising antibodies to VEEV and undetermined alphaviruses. The neurological disorder outbreak could not be linked to the alphaviruses tested. Our findings represent strong evidence that MAYV and all equine encephalitis alphaviruses circulated in the Pantanal.

  8. Neutralising antibodies for Mayaro virus in Pantanal, Brazil.

    Science.gov (United States)

    Pauvolid-Corrêa, Alex; Juliano, Raquel Soares; Campos, Zilca; Velez, Jason; Nogueira, Rita Maria Ribeiro; Komar, Nicholas

    2015-02-01

    The Pantanal hosts diverse wildlife species and therefore is a hotspot for arbovirus studies in South America. A serosurvey for Mayaro virus (MAYV), eastern (EEEV), western (WEEV) and Venezuelan (VEEV) equine encephalitis viruses was conducted with 237 sheep, 87 free-ranging caimans and 748 equids, including 37 collected from a ranch where a neurologic disorder outbreak had been recently reported. Sera were tested for specific viral antibodies using plaque-reduction neutralisation test. From a total of 748 equids, of which 264 were immunised with vaccine composed of EEEV and WEEV and 484 had no history of immunisation, 10 (1.3%) were seropositive for MAYV and two (0.3%) for VEEV using criteria of a ≥ 4-fold antibody titre difference. Among the 484 equids without history of immunisation, 48 (9.9%) were seropositive for EEEV and four (0.8%) for WEEV using the same criteria. Among the sheep, five were sero- positive for equine encephalitis alphaviruses, with one (0.4%) for EEEV, one (0.4%) for WEEV and three (1.3%) for VEEV. Regarding free-ranging caimans, one (1.1%) and three (3.4%), respectively, had low titres for neutralising antibodies to VEEV and undetermined alphaviruses. The neurological disorder outbreak could not be linked to the alphaviruses tested. Our findings represent strong evidence that MAYV and all equine encephalitis alphaviruses circulated in the Pantanal.

  9. Real-Time PCR Assay To Detect Smallpox Virus

    Science.gov (United States)

    Sofi Ibrahim, M.; Kulesh, David A.; Saleh, Sharron S.; Damon, Inger K.; Esposito, Joseph J.; Schmaljohn, Alan L.; Jahrling, Peter B.

    2003-01-01

    We developed a highly sensitive and specific assay for the rapid detection of smallpox virus DNA on both the Smart Cycler and LightCycler platforms. The assay is based on TaqMan chemistry with the orthopoxvirus hemagglutinin gene used as the target sequence. With genomic DNA purified from variola virus Bangladesh 1975, the limit of detection was estimated to be approximately 25 copies on both machines. The assay was evaluated in a blinded study with 322 coded samples that included genomic DNA from 48 different isolates of variola virus; 25 different strains and isolates of camelpox, cowpox, ectromelia, gerbilpox, herpes, monkeypox, myxoma, rabbitpox, raccoonpox, skunkpox, vaccinia, and varicella-zoster viruses; and two rickettsial species at concentrations mostly ranging from 100 fg/μl to 1 ng/μl. Contained within those 322 samples were variola virus DNA, obtained from purified viral preparations, at concentrations of 1 fg/μl to 1 ng/μl. On the Smart Cycler platform, 2 samples with false-positive results were detected among the 116 samples not containing variola virus tested; i.e., the overall specificity of the assay was 98.3%. On the LightCycler platform, five samples with false-positive results were detected (overall specificity, 95.7%). Of the 206 samples that contained variola virus DNA ranging in concentrations from 100 fg/μl to 1 ng/μl, 8 samples were considered negative on the Smart Cycler platform and 1 sample was considered negative on the LightCycler platform. Thus, the clinical sensitivities were 96.1% for the Smart Cycler instrument and 99.5% for the LightCycler instrument. The vast majority of these samples were derived from virus-infected cell cultures and variola virus-infected tissues; thus, the DNA material contained both viral DNA and cellular DNA. Of the 43 samples that contained purified variola virus DNA ranging in concentration from 1 fg/μl to 1 ng/μl, the assay correctly detected the virus in all 43 samples on both the Smart Cycler

  10. Epidemiology of hepatitis B virus infection among recyclable waste collectors in central Brazil

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    Tamíris Augusto Marinho

    2014-01-01

    Full Text Available Introduction: The collection of recyclable waste materials is a widespread activity among the urban poor. Today, this occupation attracts an increasingly large number of individuals. Despite its economic and environmental importance, this activity is associated with unsafe and unhealthy working conditions. The aim of this study was to investigate the seroepidemiological profile of hepatitis B virus (HBV infection in a population of recyclable waste collectors in central Brazil. Methods: Recyclable waste collectors from all 15 recycling cooperatives in Goiânia City were invited to participate in the study. The participants (n = 431 were interviewed and screened for hepatitis B surface antigen (HBsAg and antibodies against HBsAg (anti-HBs and hepatitis B core antigen (anti-HBc by enzyme-linked immunosorbent assay (ELISA. HBsAg- and anti-HBc-positive samples were tested for HBV DNA and genotyped. Results: The overall prevalence of HBV infection (HBsAg- and/or anti-HBc-positive was 12.8%. An age over 40 years and illicit drug use were associated with HBV infection. HBV DNA was detected in 2/3 HBsAg-positive samples and in 1/52 anti-HBc-positive/HBsAg-negative samples (an occult HBV infection rate of 1.9%, in which the genotypes/subgenotypes A/A1, D/D3 and F/F2 were identified. Only 12.3% of the recyclable waste collectors had serological evidence of previous HBV vaccination. Conclusions: These findings highlight the vulnerability of recyclable waste collectors to HBV infection and reinforce the importance of public health policies that address the health and safety of this socially vulnerable population.

  11. BLUETONGUE VIRUS ANTIBODIES DETECTIONS IN SHEEP FROM ARAÇATUBA REGION –SAO PAULO, BRAZIL DETECÇÃO DE ANTICORPOS CONTRA O VÍRUS DA LÍNGUA AZUL EM OVINOS NA REGIÃO DE ARAÇATUBA – SÃO PAULO, BRASIL

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    Adriana Hellmeister de Campos Nogueira

    2009-12-01

    Full Text Available

    Bluetongue (BT is an infectious, insect-born viral disease of ruminants. The causative agent of BT is bluetongue virus (BTV that belongs to the family Reoviridae genus Orbivirus. Insect vectors in the genus Culicoides transmit this virus. BT affects domestic and wild ruminants, however small ruminants are considered the most affected specie. The aim of the study was to detect antibodies against BTV in commercial sheep farms, of the Northeastern region of Sao Paulo State, Brazil. A total of 1002 sera samples collected from adult sheep (above 1 year-old, comprising a total of 31 farms, were screened for the presence of BTV antibodies, by agar gel immunodiffusion test (AGID and ELISA-CFS (Enzyme Linked Immunosorbent Assay – competitive solid phase, both produced by Pan American Center of FMDV. From a total of 1002 samples, 651 (65% were positive by AGID and 742 (74.1%, were positive by ELISA-CFS. These results suggest that the BTV is widespread among farms, probably causing subclinical infections.

    KEY WORDS: AGID, bluetongue virus, ELISA-CFS, seroepidemiological survey.

    A língua azul é uma doença viral, cujo agente etiológico pertence à família Reoviridae, gênero Orbivirus, transmitida por um vetor (artrópode hematófago, do gênero Culicoides. Os animais acometidos são ruminantes domésticos e selvagens, porém os pequenos ruminantes são os mais afetados. O estudo teve como objetivo detectar a presença de anticorpos para língua azul em ovinos da região de Araçatuba, por possuir um rebanho expressivo e condições climáticas favoráveis à multiplicação de insetos. Foram analisadas 1.002 amostras de soros ovinos, provenientes de 31 cabanhas, pelas provas de imunodifusão dupla em gel de ágar (AGID e ELISA (Enzyme Linked immunosorbent Assay de competição da fase sólida (ELISA CFS, provenientes do Centro Panamericano de Febre Aftosa. Desses soros, 651 (65% foram

  12. Molecular characterization of rabies virus isolated from non-haematophagous bats in Brazil

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    Avelino Albas

    2011-12-01

    Full Text Available INTRODUCTION: Rabies is an important zoonosis that causes thousands of deaths worldwide each year. Although the terrestrial cycle, mainly transmitted by dogs, is controlled in Brazil, the aerial cycle remains a serious public health issue, besides the economic problem. In the aerial cycle, the haematophagous bat Desmodus rotundus is the main source of infection, where several different species of non-haematophagous bats can be infected and can transmit the virus. METHODS: The aim of this work was to study the epidemiological pattern of rabies using antigenic characterization with monoclonal antibodies and genetic characterization by reverse-transcriptase polymerase chain reaction followed by sequencing and phylogenetic analysis of non-haematophagous bats' and herbivorous animals' central nervous system samples from the western region of the State of São Paulo, Brazil. RESULTS: From 27 samples, 3 antigenic variants were identified: AgV-3, AgV-4, and AgV-6; and from 29 samples, 5 different clusters were identified, all belonging to the rabies virus species. CONCLUSIONS: Although only non-haematophagous bats were evaluated in the studied region, the majority of samples were from antigenic and genetic variants related to haematophagous bats Desmodus rotundus. Samples from the same antigenic variant were segregated in more than one genetic cluster. This study demonstrated the diversity of rabies virus genetic lineages presented and circulating in non-haematophagous bats in the studied region.

  13. Phylogenetic analysis of rabies virus isolated from canids in North and Northeast Brazil.

    Science.gov (United States)

    de Souza, Débora Nunes; Carnieli, Pedro; Macedo, Carla Isabel; de Novaes Oliveira, Rafael; de Carvalho Ruthner Batista, Helena Beatriz; Rodrigues, Adriana Candido; Pereira, Patricia Mariano Cruz; Achkar, Samira Maria; Vieira, Luiz Fernando Pereira; Kawai, Juliana Galera Castilho

    2017-01-01

    Cases of canine rabies continue to occur in North and Northeast Brazil, and the number of notifications of rabies cases in wild canids has increased as a result of the expansion of urban areas at the expense of areas with native vegetation. In light of this, we performed molecular characterization of rabies virus isolates from dogs and Cerdocyon thous from various states in North and Northeast Brazil. In all, 102 samples from dogs (n = 56) and Cerdocyon thous (n = 46) collected between 2006 and 2012 were used. The nucleotide sequences obtained for the N gene of rabies virus were analyzed, and phylogenetic analysis revealed the presence of two distinct genetic lineages, one associated with canids and one with bats, and, within the canid cluster, two distinct sublineages circulating among dogs and Cerdocyon thous. In addition, phylogenetic groups associated with geographic region and fourteen cases of interspecific infection were observed among the isolates from canids. Our findings show that analysis of rabies virus lineages isolated from reservoirs such as canids must be constantly evaluated because the mutation rate is high.

  14. Vaccinia Virus Natural Infections in Brazil: The Good, the Bad, and the Ugly

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    Jaqueline Silva de Oliveira

    2017-11-01

    Full Text Available The orthopoxviruses (OPV comprise several emerging viruses with great importance to human and veterinary medicine, including vaccinia virus (VACV, which causes outbreaks of bovine vaccinia (BV in South America. Historically, VACV is the most comprehensively studied virus, however, its origin and natural hosts remain unknown. VACV was the primary component of the smallpox vaccine, largely used during the smallpox eradication campaign. After smallpox was declared eradicated, the vaccination that conferred immunity to OPV was discontinued, favoring a new contingent of susceptible individuals to OPV. VACV infections occur naturally after direct contact with infected dairy cattle, in recently vaccinated individuals, or through alternative routes of exposure. In Brazil, VACV outbreaks are frequently reported in rural areas, affecting mainly farm animals and humans. Recent studies have shown the role of wildlife in the VACV transmission chain, exploring the role of wild rodents as reservoirs that facilitate VACV spread throughout rural areas. Furthermore, VACV circulation in urban environments and the significance of this with respect to public health, have also been explored. In this review, we discuss the history, epidemiological, ecological and clinical aspects of natural VACV infections in Brazil, also highlighting alternative routes of VACV transmission, the factors involved in susceptibility to infection, and the natural history of the disease in humans and animals, and the potential for dissemination to urban environments.

  15. Avian influenza virus (H11N9 in migratory shorebirds wintering in the Amazon Region, Brazil.

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    Jansen de Araujo

    Full Text Available Aquatic birds are the natural reservoir for avian influenza viruses (AIV. Habitats in Brazil provide stopover and wintering sites for water birds that migrate between North and South America. The current study was conducted to elucidate the possibility of the transport of influenza A viruses by birds that migrate annually between the Northern and Southern Hemispheres. In total, 556 orotracheal/cloacal swab samples were collected for influenza A virus screening using real-time RT-PCR (rRT-PCR. The influenza A virus-positive samples were subjected to viral isolation. Four samples were positive for the influenza A matrix gene by rRT-PCR. From these samples, three viruses were isolated, sequenced and characterized. All positive samples originated from a single bird species, the ruddy turnstone (Arenaria interpres, that was caught in the Amazon region at Caeté Bay, Northeast Pará, at Ilha de Canelas. To our knowledge, this is the first isolation of H11N9 in the ruddy turnstone in South America.

  16. Avian Influenza Virus (H11N9) in Migratory Shorebirds Wintering in the Amazon Region, Brazil

    Science.gov (United States)

    de Araujo, Jansen; de Azevedo Júnior, Severino M.; Gaidet, Nicolas; Hurtado, Renata F.; Walker, David; Thomazelli, Luciano M.; Ometto, Tatiana; Seixas, Marina M. M.; Rodrigues, Roberta; Galindo, Daniele B.; da Silva, Adriana C. S.; Rodrigues, Arlinéa M. M.; Bomfim, Leonardo L.; Mota, Marcelo A.; Larrazábal, Maria E.; Branco, Joaquim O.; Serafini, Patricia; Neto, Isaac S.; Franks, John; Webby, Richard J.; Webster, Robert G.; Durigon, Edison L.

    2014-01-01

    Aquatic birds are the natural reservoir for avian influenza viruses (AIV). Habitats in Brazil provide stopover and wintering sites for water birds that migrate between North and South America. The current study was conducted to elucidate the possibility of the transport of influenza A viruses by birds that migrate annually between the Northern and Southern Hemispheres. In total, 556 orotracheal/cloacal swab samples were collected for influenza A virus screening using real-time RT-PCR (rRT-PCR). The influenza A virus-positive samples were subjected to viral isolation. Four samples were positive for the influenza A matrix gene by rRT-PCR. From these samples, three viruses were isolated, sequenced and characterized. All positive samples originated from a single bird species, the ruddy turnstone (Arenaria interpres), that was caught in the Amazon region at Caeté Bay, Northeast Pará, at Ilha de Canelas. To our knowledge, this is the first isolation of H11N9 in the ruddy turnstone in South America. PMID:25329399

  17. Molecular characterization of hepatitis B virus genotype A from Argentina and Brazil.

    Science.gov (United States)

    Mbayed, Viviana Andrea; Piñeiro y Leone, Flavia Guadalupe; Pezzano, Silvana Claudia; Campos, Rodolfo Héctor

    2009-01-01

    To study the diversification of hepatitis B virus genotype A in Latin America, we analyzed seven new Argentinian isolates and published sequences from Argentina and Brazil and other countries from the region. We found that the European subgenotype A2 prevailed in most of the countries except for Brazil, where the African subgenotype A1 predominated. A2 isolates did not differ significantly from the GenBank sequences, whereas some A1 isolates carried, concomitantly, amino acids characteristic of the subgenotypes A3 (R(501) in P protein) and A2 (D(355) in P/T(54) in preS2). This combination is absent in the A1 subgenotype around the world. We discuss the origin, distribution and introduction of those subgenotypes in the Americas.

  18. Characterization of Aujeszky's disease virus isolates from South and Southeast Brazil by RFLP analysis

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    Piatti Rosa Maria

    2001-01-01

    Full Text Available The genomic DNA of thirty strains of Aujeszky's disease virus (ADV isolated in the South and Southeast regions of Brazil from 1982 to 1996 were characterized by restriction endonuclease analysis with BamHI. Twenty seven strains were isolated from pigs, 1 from cattle, 1 from cat and 1 from dog. Using a systematization previously described, the 30 ADV strains could be classified as genomic types I (n = 2 and II (n = 28. Genomic type III was not observed. In this first study of genomic type characterization of brazilian ADV strains, we could demonstrate the occurence in Brazil of the genomic types I and II, with a large predominance of genomic type II.

  19. Serological markers and risk factors related to hepatitis B virus in dentists in the Central West region of Brazil

    Science.gov (United States)

    de Paiva, Enilza Maria Mendonça; Tiplle, Anaclara Ferreira Veiga; de Paiva Silva, Eliane; de Paula Cardoso, Divina das Dores

    2008-01-01

    The hepatitis B virus (HBV) has been considered the major occupational risk agent for dentists. The Central West region of Brazil is considered an intermediate endemic pattern area, but currently there is no information about the HBV prevalence in dentists of Goiânia, Goiás. This study aimed at the detection of the HBV infection rate and risk factors for dentists of Goiânia and the comparison of the obtained data with the general population and other groups. A randomized sample of 680 professionals participated in this study. All dentists gave written consent for the procedure and filled out a questionnaire about risk factors. The HBV serological markers were analyzed using ELISA test and the presence of anti-HBc was observed in 41 (6.0%) of the dentists. None of them was HBsAg positive. Significant relationships with HBV positivity were observed with gender, the time working as a dentist and the use of incomplete personal protective equipment (PPE). The HBV prevalence found in this group of dentists was lower than the endemic pattern of the general population, other health care workers of the region and the dentists from other regions in Brazil. These results may indicate a positive impact of vaccination considering the high adherence of the dentists to the immunization program (98.4%). Finally, the use of complete PPE by the majority as well as other standard precautions recommended for health care workers could be responsible for the low HBV seroprevalence. PMID:24031211

  20. Identification of novel recombinants of hepatitis B virus genotypes F and G in human immunodeficiency virus-positive patients from Argentina and Brazil.

    Science.gov (United States)

    Araujo, Natalia M; Araujo, Oscar C; Silva, Edinete M; Villela-Nogueira, Cristiane A; Nabuco, Letícia C; Parana, Raymundo; Bessone, Fernando; Gomes, Selma A; Trepo, Christian; Kay, Alan

    2013-01-01

    Hepatitis B virus (HBV) genotype G (HBV/G) infection is almost always detected along with a co-infecting HBV strain that can supply HBeAg, typically HBV/A2. In this study we describe, in two human immunodeficiency virus (HIV)-positive patients from Argentina and Brazil, the first report of HBV/G infection in Argentina and co-circulation of HBV/G, HBV/F and G/F recombinants in the American continent. HBV isolates carrying the 36 bp insertion of HBV/G were the most prevalent in both patients, with >99 % of colonies hybridizing to a probe specific for this insertion. Phylogenetic analyses of full-length genomes and precore/core fragments revealed that F4 and F1b were the co-infecting subgenotypes in the Brazilian and Argentinian patients, respectively. Bootscanning analysis provided evidence of recombination in several clones from both patients, with recombination breakpoints located mainly at the precore/core region. These data should encourage further investigations on the clinical implications of HBV/G recombinants in HBV/HIV co-infected patients.

  1. Prevalence of hepatitis C Virus infection among hemophiliacs in Central Brazil

    Directory of Open Access Journals (Sweden)

    Adriana P Barbosa

    2002-07-01

    Full Text Available In order to investigate the hepatitis C virus (HCV infection prevalence and risk factors in hemophiliacs in Central Brazil, 90 patients were interviewed and serum samples tested for HCV RNA and anti-HCV antibodies. An overall prevalence of 63.3% (CI 95%: 53.0-72.7 was found. Multivariate analysis of risk factors showed that number of blood transfusions was significantly associated with this infection. Most hemophiliacs received locally produced cryoprecipitate. All infected patients were transfused before the screening of blood units for anti-HCV. However, hemophiliacs who received exclusively screened cryoprecipitate were HCV negative. It confirms the expected decline in transfusion-acquired hepatitis C.

  2. Detecting RNA viruses in living mammalian cells by fluorescence microscopy.

    Science.gov (United States)

    Sivaraman, Divya; Biswas, Payal; Cella, Lakshmi N; Yates, Marylynn V; Chen, Wilfred

    2011-07-01

    Traditional methods that rely on viral isolation and culture techniques continue to be the gold standards used for detection of infectious viral particles. However, new techniques that rely on visualization of live cells can shed light on understanding virus-host interaction for early stage detection and potential drug discovery. Live-cell imaging techniques that incorporate fluorescent probes into viral components provide opportunities for understanding mRNA expression, interaction, and virus movement and localization. Other viral replication events inside a host cell can be exploited for non-invasive detection, such as single-virus tracking, which does not inhibit viral infectivity or cellular function. This review highlights some of the recent advances made using these novel approaches for visualization of viral entry and replication in live cells. Copyright © 2011 Elsevier Ltd. All rights reserved.

  3. Presence and Distribution of Oilseed Pumpkin Viruses and Molecular Detection of Zucchini Yellow Mosaic Virus

    Directory of Open Access Journals (Sweden)

    Ana Vučurović

    2009-01-01

    Full Text Available Over the past decade, intensive spread of virus infections of oilseed pumpkin has resulted in significant economic losses in pumpkin crop production, which is currently expanding in our country. In 2007 and 2008, a survey for the presence and distribution of oilseed pumpkin viruses was carried out in order to identify viruses responsible for epidemics and incidences of very destructive symptoms on cucurbit leaves and fruits. Monitoring andcollecting samples of oil pumpkin, as well as other species such as winter and butternut squash and buffalo and bottle gourd with viral infection symptoms, was conducted in several localities of Vojvodina Province. The collected plant samples were tested by DAS-ELISA using polyclonal antisera specific for the detection of six most economically harmful pumpkin viruses: Cucumber mosaic virus (CMV, Zucchini yellow mosaic virus (ZYMV, Watermelon mosaic virus (WMW, Squash mosaic virus (SqMV, Papaya ringspot virus (PRSV and Tobaccoringspot virus (TRSV that are included in A1 quarantine list of harmful organisms in Serbia.Identification of viruses in the collected samples indicated the presence of three viruses, ZYMV, WMV and CMV, in individual and mixed infections. Frequency of the identified viruses varied depending on locality and year of investigations. In 2007, WMV was the most frequent virus (94.2%, while ZYMV was prevalent (98.04% in 2008. High frequency of ZYMV determined in both years of investigation indicated the need for its rapid and reliable molecular detection. During this investigation, a protocol for ZYMVdetection was developed and optimized using specific primers CPfwd/Cprev and commercial kits for total RNA extraction, as well as for RT-PCR. In RT-PCR reaction using these primers, a DNA fragment of approximately 1100 bp, which included coat protein gene, was amplified in the samples of infected pumkin leaves. Although serological methods are still useful for large-scale testing of a great number of

  4. The SIR model of Zika virus disease outbreak in Brazil at year 2015

    Science.gov (United States)

    Aik, Lim Eng; Kiang, Lam Chee; Hong, Tan Wei; Abu, Mohd Syafarudy

    2017-05-01

    This research study demonstrates a numerical model intended for comprehension the spread of the year 2015 Zika virus disease utilizing the standard SIR framework. In modeling virulent disease dynamics, it is important to explore whether the illness spread could accomplish a pandemic level or it could be eradicated. Information from the year 2015 Zika virus disease event is utilized and Brazil where the event began is considered in this research study. A three dimensional nonlinear differential equation is formulated and solved numerically utilizing the Euler's method in MS excel. It is appeared from the research study that, with health intercessions of public, the viable regenerative number can be decreased making it feasible for the event to cease to exist. It is additionally indicated numerically that the pandemic can just cease to exist when there are no new infected people in the populace.

  5. Single Assay Detection of Acute Bee Paralysis Virus, Kashmir Bee Virus and Israeli Acute Paralysis Virus

    DEFF Research Database (Denmark)

    Francis, Roy Mathew; Kryger, Per

    2012-01-01

    A new RT-PCR primer pair designed to identify Acute Bee Paralysis Virus (ABPV), Kashmir Bee Virus (KBV) or Israeli Acute Bee Paralysis Virus (IAPV) of honey bees (Apis mellifera L.) in a single assay is described. These primers are used to screen samples for ABPV, KBV, or IAPV in a single RT...

  6. A novel, highly divergent ssDNA virus identified in Brazil infecting apple, pear and grapevine.

    Science.gov (United States)

    Basso, Marcos Fernando; da Silva, José Cleydson Ferreira; Fajardo, Thor Vinícius Martins; Fontes, Elizabeth Pacheco Batista; Zerbini, Francisco Murilo

    2015-12-02

    Fruit trees of temperate and tropical climates are of great economical importance worldwide and several viruses have been reported affecting their productivity and longevity. Fruit trees of different Brazilian regions displaying virus-like symptoms were evaluated for infection by circular DNA viruses. Seventy-four fruit trees were sampled and a novel, highly divergent, monopartite circular ssDNA virus was cloned from apple, pear and grapevine trees. Forty-five complete viral genomes were sequenced, with a size of approx. 3.4 kb and organized into five ORFs. Deduced amino acid sequences showed identities in the range of 38% with unclassified circular ssDNA viruses, nanoviruses and alphasatellites (putative Replication-associated protein, Rep), and begomo-, curto- and mastreviruses (putative coat protein, CP, and movement protein, MP). A large intergenic region contains a short palindromic sequence capable of forming a hairpin-like structure with the loop sequence TAGTATTAC, identical to the conserved nonanucleotide of circoviruses, nanoviruses and alphasatellites. Recombination events were not detected and phylogenetic analysis showed a relationship with circo-, nano- and geminiviruses. PCR confirmed the presence of this novel ssDNA virus in field plants. Infectivity tests using the cloned viral genome confirmed its ability to infect apple and pear tree seedlings, but not Nicotiana benthamiana. The name "Temperate fruit decay-associated virus" (TFDaV) is proposed for this novel virus. Copyright © 2015 Elsevier B.V. All rights reserved.

  7. High Zika Virus Seroprevalence in Salvador, Northeastern Brazil Limits the Potential for Further Outbreaks

    Science.gov (United States)

    Netto, Eduardo Martins; Moreira-Soto, Andres; Pedroso, Celia; Höser, Christoph; Funk, Sebastian; Kucharski, Adam J.; Rockstroh, Alexandra; Kümmerer, Beate M.; Sampaio, Gilmara Souza; Luz, Estela; Vaz, Sara Nunes; Dias, Juarez Pereira; Bastos, Fernanda Anjos; Cabral, Renata; Kistemann, Thomas; Ulbert, Sebastian; de Lamballerie, Xavier; Jaenisch, Thomas; Brady, Oliver J.; Drosten, Christian; Sarno, Manoel

    2017-01-01

    ABSTRACT During 2015 to 2016, Brazil reported more Zika virus (ZIKV) cases than any other country, yet population exposure remains unknown. Serological studies of ZIKV are hampered by cross-reactive immune responses against heterologous viruses. We conducted serosurveys for ZIKV, dengue virus (DENV), and Chikungunya virus (CHIKV) in 633 individuals prospectively sampled during 2015 to 2016, including microcephaly and non-microcephaly pregnancies, HIV-infected patients, tuberculosis patients, and university staff in Salvador in northeastern Brazil using enzyme-linked immunosorbent assays (ELISAs) and plaque reduction neutralization tests. Sera sampled retrospectively during 2013 to 2015 from 277 HIV-infected patients were used to assess the spread of ZIKV over time. Individuals were georeferenced, and sociodemographic indicators were compared between ZIKV-positive and -negative areas and areas with and without microcephaly cases. Epidemiological key parameters were modeled in a Bayesian framework. ZIKV seroprevalence increased rapidly during 2015 to 2016, reaching 63.3% by 2016 (95% confidence interval [CI], 59.4 to 66.8%), comparable to the seroprevalence of DENV (75.7%; CI, 69.4 to 81.1%) and higher than that of CHIKV (7.4%; CI, 5.6 to 9.8%). Of 19 microcephaly pregnancies, 94.7% showed ZIKV IgG antibodies, compared to 69.3% of 257 non-microcephaly pregnancies (P = 0.017). Analyses of sociodemographic data revealed a higher ZIKV burden in low socioeconomic status (SES) areas. High seroprevalence, combined with case data dynamics allowed estimates of the basic reproduction number R0 of 2.1 (CI, 1.8 to 2.5) at the onset of the outbreak and an effective reproductive number Reff of ZIKV-associated congenital disease and an association of low SES and ZIKV infection and suggest that population immunity caused cessation of the outbreak. Similar studies from other areas will be required to determine the fate of the American ZIKV outbreak. PMID:29138300

  8. Detection of chikungunya virus in saliva and urine.

    Science.gov (United States)

    Musso, Didier; Teissier, Anita; Rouault, Eline; Teururai, Sylviane; de Pina, Jean-Jacques; Nhan, Tu-Xuan

    2016-06-16

    Saliva and urine have been used for arthropod-borne viruses molecular detection but not yet for chikungunya virus (CHIKV). We investigated the use of saliva and urine for molecular detection of CHIKV during the French Polynesian outbreak. During the French Polynesian chikungunya outbreak (2014-2015), we collected the same day blood and saliva samples from 60 patients with probable chikungunya (47 during the 1st week post symptoms onset and 13 after), urine was available for 39 of them. All samples were tested using a CHIKV reverse-transcription PCR. Forty eight patients had confirmed chikungunya. For confirmed chikungunya presenting during the 1st week post symptoms onset, CHIKV RNA was detected from 86.1 % (31/36) of blood, 58.3 % (21/36) of saliva and 8.3 % (2/24) of urine. Detection rate of CHIKV RNA was significantly higher in blood compared to saliva. For confirmed chikungunya presenting after the 1st week post symptoms onset, CHIKV RNA was detected from 8.3 % (1/12) of blood, 8.3 % (1/12) of saliva and 0 % (0/8) of urine. In contrast to Zika virus (ZIKV), saliva did not increased the detection rate of CHIKV RNA during the 1st week post symptoms onset. In contrast to ZIKV, dengue virus and West Nile virus, urine did not enlarged the window of detection of CHIKV RNA after the 1st week post symptoms onset. Saliva can be used for molecular detection of CHIKV during the 1st week post symptoms onset only if blood is impossible to collect but with a lower sensitivity compared to blood.

  9. Plum pox virus variability detected by the advanced analytical methods.

    Science.gov (United States)

    Subr, Z; Glasa, M

    2008-01-01

    Plum pox virus (PPV) infects stone-fruit trees with important economical impact mainly in Europe and Mediterranean region. The data about PPV intra-species variability accumulated markedly in the last two decades. Six PPV strains have been recognized using different approaches including serology, protein analysis, specific amplification, and genome sequencing. Reliable and sensitive diagnostics is the most important requirement for application of early control and safety measures. Therefore, many techniques and their modifications have been adapted to detect PPV and its different forms. Here, we review the improvement of the PPV detection and variability analysis in the context of progress in laboratory methods since the virus discovery till today.

  10. Detection and analysis of the Chameleon WiFi access point virus

    National Research Council Canada - National Science Library

    Milliken, Jonny; Selis, Valerio; Marshall, Alan

    2013-01-01

    This paper analyses and proposes a novel detection strategy for the 'Chameleon’ WiFi AP-AP virus. Previous research has considered virus construction, likely virus behaviour and propagation methods...

  11. Easy and Rapid Detection of Mumps Virus by Live Fluorescent Visualization of Virus-Infected Cells.

    Science.gov (United States)

    Takahashi, Tadanobu; Agarikuchi, Takashi; Kurebayashi, Yuuki; Shibahara, Nona; Suzuki, Chihiro; Kishikawa, Akiko; Fukushima, Keijo; Takano, Maiko; Suzuki, Fumie; Wada, Hirohisa; Otsubo, Tadamune; Ikeda, Kiyoshi; Minami, Akira; Suzuki, Takashi

    2015-01-01

    Mumps viruses show diverse cytopathic effects (CPEs) of infected cells and viral plaque formation (no CPE or no plaque formation in some cases) depending on the viral strain, highlighting the difficulty in mumps laboratory studies. In our previous study, a new sialidase substrate, 2-(benzothiazol-2-yl)-4-bromophenyl 5-acetamido-3,5-dideoxy-α-D-glycero-D-galacto-2-nonulopyranosidonic acid (BTP3-Neu5Ac), was developed for visualization of sialidase activity. BTP3-Neu5Ac can easily and rapidly perform histochemical fluorescent visualization of influenza viruses and virus-infected cells without an antiviral antibody and cell fixation. In the present study, the potential utility of BTP3-Neu5Ac for rapid detection of mumps virus was demonstrated. BTP3-Neu5Ac could visualize dot-blotted mumps virus, virus-infected cells, and plaques (plaques should be called focuses due to staining of infected cells in this study), even if a CPE was not observed. Furthermore, virus cultivation was possible by direct pick-up from a fluorescent focus. In conventional methods, visible appearance of the CPE and focuses often requires more than 6 days after infection, but the new method with BTP3-Neu5Ac clearly visualized infected cells after 2 days and focuses after 4 days. The BTP3-Neu5Ac assay is a precise, easy, and rapid assay for confirmation and titration of mumps virus.

  12. Easy and Rapid Detection of Mumps Virus by Live Fluorescent Visualization of Virus-Infected Cells.

    Directory of Open Access Journals (Sweden)

    Tadanobu Takahashi

    Full Text Available Mumps viruses show diverse cytopathic effects (CPEs of infected cells and viral plaque formation (no CPE or no plaque formation in some cases depending on the viral strain, highlighting the difficulty in mumps laboratory studies. In our previous study, a new sialidase substrate, 2-(benzothiazol-2-yl-4-bromophenyl 5-acetamido-3,5-dideoxy-α-D-glycero-D-galacto-2-nonulopyranosidonic acid (BTP3-Neu5Ac, was developed for visualization of sialidase activity. BTP3-Neu5Ac can easily and rapidly perform histochemical fluorescent visualization of influenza viruses and virus-infected cells without an antiviral antibody and cell fixation. In the present study, the potential utility of BTP3-Neu5Ac for rapid detection of mumps virus was demonstrated. BTP3-Neu5Ac could visualize dot-blotted mumps virus, virus-infected cells, and plaques (plaques should be called focuses due to staining of infected cells in this study, even if a CPE was not observed. Furthermore, virus cultivation was possible by direct pick-up from a fluorescent focus. In conventional methods, visible appearance of the CPE and focuses often requires more than 6 days after infection, but the new method with BTP3-Neu5Ac clearly visualized infected cells after 2 days and focuses after 4 days. The BTP3-Neu5Ac assay is a precise, easy, and rapid assay for confirmation and titration of mumps virus.

  13. Assay optimization for molecular detection of Zika virus

    Science.gov (United States)

    Corman, Victor M; Rasche, Andrea; Baronti, Cecile; Aldabbagh, Souhaib; Cadar, Daniel; Reusken, Chantal BEM; Pas, Suzan D; Goorhuis, Abraham; Schinkel, Janke; Molenkamp, Richard; Kümmerer, Beate M; Bleicker, Tobias; Brünink, Sebastian; Eschbach-Bludau, Monika; Eis-Hübinger, Anna M; Koopmans, Marion P; Schmidt-Chanasit, Jonas; Grobusch, Martin P; de Lamballerie, Xavier; Drosten, Christian

    2016-01-01

    Abstract Objective To examine the diagnostic performance of real-time reverse transcription (RT)-polymerase chain reaction (PCR) assays for Zika virus detection. Methods We compared seven published real-time RT–PCR assays and two new assays that we have developed. To determine the analytical sensitivity of each assay, we constructed a synthetic universal control ribonucleic acid (uncRNA) containing all of the assays’ target regions on one RNA strand and spiked human blood or urine with known quantities of African or Asian Zika virus strains. Viral loads in 33 samples from Zika virus-infected patients were determined by using one of the new assays. Findings Oligonucleotides of the published real-time RT–PCR assays, showed up to 10 potential mismatches with the Asian lineage causing the current outbreak, compared with 0 to 4 mismatches for the new assays. The 95% lower detection limit of the seven most sensitive assays ranged from 2.1 to 12.1 uncRNA copies/reaction. Two assays had lower sensitivities of 17.0 and 1373.3 uncRNA copies/reaction and showed a similar sensitivity when using spiked samples. The mean viral loads in samples from Zika virus-infected patients were 5 × 104 RNA copies/mL of blood and 2 × 104 RNA copies/mL of urine. Conclusion We provide reagents and updated protocols for Zika virus detection suitable for the current outbreak strains. Some published assays might be unsuitable for Zika virus detection, due to the limited sensitivity and potential incompatibility with some strains. Viral concentrations in the clinical samples were close to the technical detection limit, suggesting that the use of insensitive assays will cause false-negative results. PMID:27994281

  14. Serological evidence for Saint Louis encephalitis virus in free-ranging New World monkeys and horses within the upper Paraná River basin region, Southern Brazil

    Directory of Open Access Journals (Sweden)

    Walfrido Kühl Svoboda

    2014-06-01

    Full Text Available Introduction Saint Louis encephalitis virus (SLEV primarily occurs in the Americas and produces disease predominantly in humans. This study investigated the serological presence of SLEV in nonhuman primates and horses from southern Brazil. Methods From June 2004 to December 2005, sera from 133 monkeys (Alouatta caraya, n=43; Sapajus nigritus, n=64; Sapajus cay, n=26 trap-captured at the Paraná River basin region and 23 blood samples from farm horses were obtained and used for the serological detection of a panel of 19 arboviruses. All samples were analyzed in a hemagglutination inhibition (HI assay; positive monkey samples were confirmed in a mouse neutralization test (MNT. Additionally, all blood samples were inoculated into C6/36 cell culture for viral isolation. Results Positive seroreactivity was only observed for SLEV. A prevalence of SLEV antibodies in sera was detected in Alouatta caraya (11.6%; 5/43, Sapajus nigritus (12.5%; 8/64, and S. cay (30.8%; 8/26 monkeys with the HI assay. Of the monkeys, 2.3% (1/42 of A. caraya, 6.3% 94/64 of S. nigritus, and 15.4% (4/26 of S. cay were positive for SLEV in the MNT. Additionally, SLEV antibodies were detected by HI in 39.1% (9/23 of the horses evaluated in this study. Arboviruses were not isolated from any blood sample. Conclusions These results confirmed the presence of SLEV in nonhuman primates and horses from southern Brazil. These findings most likely represent the first detection of this virus in nonhuman primates beyond the Amazon region. The detection of SLEV in animals within a geographical region distant from the Amazon basin suggests that there may be widespread and undiagnosed dissemination of this disease in Brazil.

  15. Susceptibility of urban and rural populations of Aedes albopictus from São Paulo State, Brazil, to infection by dengue-1 and -2 viruses.

    Science.gov (United States)

    Fernández, Zoraida; Moncayo, Abelardo; Forattini, Oswaldo Paulo; Weaver, Scott C

    2004-09-01

    In Brazil, dengue viruses (DENV) are transmitted by Aedes aegypti (L.). However, the recent introduction of Aedes albopictus (Skuse) has caused concern, because some populations of this species also are capable of DENV transmission. Ae. albopictus from rural and urban localities of São Paulo State, Brazil, were artificially exposed to infection with DENV-1 and -2 to evaluate their susceptibility. The rates of infection (6.2 and 5.7% for DENV-1; 5.3 and 12% for DENV-2) and dissemination (22.2 and 40% for DENV-1; 0 and 20% for DENV-2) were low in both urban and rural populations, with no significant differences detected between these mosquito populations.

  16. Evidence of Zika Virus RNA Fragments in Aedes albopictus (Diptera: Culicidae) Field-Collected Eggs From Camaçari, Bahia, Brazil.

    Science.gov (United States)

    Smartt, Chelsea T; Stenn, Tanise M S; Chen, Tse-Yu; Teixeira, Maria Gloria; Queiroz, Erivaldo P; Souza Dos Santos, Luciano; Queiroz, Gabriel A N; Ribeiro Souza, Kathleen; Kalabric Silva, Luciano; Shin, Dongyoung; Tabachnick, Walter J

    2017-07-01

    A major mosquito-borne viral disease outbreak caused by Zika virus (ZIKV) occurred in Bahia, Brazil, in 2015, largely due to transmission by the mosquito, Aedes aegypti (L.). Detecting ZIKV in field samples of Ae. aegypti has proven problematic in some locations, suggesting other mosquito species might be contributing to the spread of ZIKV. In this study, several (five) adult Aedes albopictus (Skuse) mosquitoes that emerged from a 2015 field collection of eggs from Camaçari, Bahia, Brazil, were positive for ZIKV RNA; however, attempts to isolate live virus were not successful. Results from this study suggest that field-collected Ae. albopictus eggs may contain ZIKV RNA that require further tests for infectious ZIKV. There is a need to investigate the role of Ae. albopictus in the ZIKV infection process in Brazil and to study the potential presence of vertical and sexual transmission of ZIKV in this species. © The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  17. Aptasensor development for detection of virus in water

    DEFF Research Database (Denmark)

    Kirkegaard, Julie

    Contamination of water by waterborne viruses causes serious health issues worldwide. The current virus detection methods are expensive and time-consuming and require access to well-equipped laboratories. This thesis describes the development of an impedimetric all-polymer aptasensor for detection...... and finally an impedimetric electrical characterisation of the sensor. The sample preparation of the rotavirus was based on purification and biotinylation of the virus to meet the requirements of the aptamer selection process. The selection process, performed by an external collaborator, was based...... was stable in the measured pH range, which corresponds to the expected pH range of water samples.. The stability of the aptasensor chip was tested over a 2 week period in continuous flow. It was found that the electrodes were not damaged or degraded during the time period, as a constant impedance signal...

  18. Integrated optic immunoassay for virus detection

    Science.gov (United States)

    Boiarski, Anthony A.; Busch, James R.; Miller, Larry S.; Zulich, A. W.; Burans, James

    1995-05-01

    An integrated optic refractometer device was developed to perform a rapid one-step, label-free immunoassay. The device measures refractive index changes at the surface of a planar waveguide using interferometry. Antibodies were applied to the waveguide surface to provide a bioselective coating for detecting and quantifying a specific antigen of interest. The detection limit of this biosensor was determined for adenovirus as a model for other viral analytes of military, medical, and environmental interest. As binding of the antigen occurred on the sensor surface, a time-dependent phase shift of the helium-neon laser light beam was detected and was measured over a 10-minute time period. Adenovirus was detected at levels of 250 - 2500 viral particles/ml. This detection limit was obtained for a mono-layer of antibody attached to the sensor. Use of a high-density, multi-layer antibody coating approach resulted in improved detection limits for bacteria and protein analytes of general interest.

  19. Assay optimization for molecular detection of Zika virus

    NARCIS (Netherlands)

    Corman, Victor M.; Rasche, Andrea; Baronti, Cecile; Aldabbagh, Souhaib; Cadar, Daniel; Reusken, Chantal Bem; Pas, Suzan D.; Goorhuis, Abraham; Schinkel, Janke; Molenkamp, Richard; Kümmerer, Beate M.; Bleicker, Tobias; Brünink, Sebastian; Eschbach-Bludau, Monika; Eis-Hübinger, Anna M.; Koopmans, Marion P.; Schmidt-Chanasit, Jonas; Grobusch, Martin P.; de Lamballerie, Xavier; Drosten, Christian; Drexler, Jan Felix

    2016-01-01

    To examine the diagnostic performance of real-time reverse transcription (RT)-polymerase chain reaction (PCR) assays for Zika virus detection. We compared seven published real-time RT-PCR assays and two new assays that we have developed. To determine the analytical sensitivity of each assay, we

  20. Detection of pepper leaf curl virus through PCR amplification and ...

    African Journals Online (AJOL)

    Pepper leaf curl virus (PepLCV) is the most destructive pathogen of pepper and causes substantial economic losses of chilli production worldwide. Curling and puckering of leaves and stunted growth of the plants are typical symptoms of the viral infection. For a reliable detection of PepLCV, coat protein specific primer pairs ...

  1. Serological Detection of Infectious Bursa Disease Virus Antibodies ...

    African Journals Online (AJOL)

    The detection and distribution of infectious Bursa disease (IBD) virus antibody among local species of birds was investigated in 4,655 sera sample using Agar Gel precipitation test (AGPT). The results showed that local chickens had the highest distribution with 446 (9.58%) followed by ducks 218 (4.68%), guinea fowl 131 ...

  2. Molecular and serological detection of occult hepatitis B virus ...

    African Journals Online (AJOL)

    Background: Occult hepatitis B infections are becoming a major global threat, but the available data on its prevalence in various parts of the world are often divergent. Objective: This study aimed to detect occult hepatitis B virus in hepatitis B surface antigen-negative serum using anti-HBc as a marker of previous infection.

  3. Molecular and serological detection of occult hepatitis B virus ...

    African Journals Online (AJOL)

    Molecular and serological detection of occult hepatitis B virus among healthy hepatitis B surface ... Journal Home > Vol 16, No 3 (2016) > ... The PDF file you selected should load here if your Web browser has a PDF reader plug-in installed ...

  4. Detection of viruses in seeds of some vegetables by reverse ...

    African Journals Online (AJOL)

    my mord

    The present study was carried out to detect the presence of Alfalfa mosaic alfamovirus (AMV), cucumber mosaic cucumovirus (CMV), lettuce mosaic potyvirus (LMV), cucumber green mottle mosaic virus (CGMMV), tomato bushy stunt tombusvirus (TBSV), tobacco mosaic tobamovirus (TMV), tomato black ring nepovirus ...

  5. Stable recombinant alpaca antibodies for detection of Tulip virus X

    NARCIS (Netherlands)

    Beekwilder, M.J.; Houwelingen, van A.M.M.L.; Beckhoven, van J.R.C.M.; Speksnijder, A.G.C.L.

    2008-01-01

    For detection of the plant pathogenic Tulip virus X (TuVX), a panel of six recombinant antibodies was identified. To this end, a repertoire of variable domains from heavy-chain immunoglobulins (VHH) was cloned from an alpaca, which had been immunized with TuVX. Binding domains were selected by phage

  6. Serological Detection of Infectious Bursa Disease Virus Antibodies ...

    African Journals Online (AJOL)

    The detection and distribution of infectious Bursa disease (IBD) virus antibody among local species of birds was investigated in 4,655 ... imply that the birds have the potentials of becoming carriers and serve as source of spread of the disease to other ..... In Nigeria we have some militating factors against the infectious bursa ...

  7. Serological detection of seed borne viruses in cowpea regenerated ...

    African Journals Online (AJOL)

    ... conservation and exchange between countries as it causes great loss. Southern bean mosaic virus was not detected in any of the accessions evaluated. Seeds were harvested from the infected mother plants at maturity and the embryos excised in vitro onto hormone free medium composed of 4.43 g MS basal salt, 30 g ...

  8. Assay optimization for molecular detection of Zika virus

    NARCIS (Netherlands)

    Corman, Victor M.; Rasche, Andrea; Baronti, Cecile; Aldabbagh, Souhaib; Cadar, Daniel; Reusken, Chantal Bem; Pas, Suzan D.; Goorhuis, Abraham; Schinkel, Janke; Molenkamp, Richard; Kümmerer, Beate M.; Bleicker, Tobias; Brünink, Sebastian; Eschbach-Bludau, Monika; Eis-Hübinger, Anna M.; Koopmans, Marion P.; Schmidt-Chanasit, Jonas; Grobusch, Martin P.; de Lamballerie, Xavier; Drosten, Christian; Drexler, Jan Felix

    2016-01-01

    Objective To examine the diagnostic performance of real-time reverse transcription (RT)-polymerase chain reaction (PCR) assays for Zika virus detection. Methods We compared seven published real-time RT PCR assays and two new assays that we have developed. To determine the analytical sensitivity of

  9. A monoclonal blocking ELISA to detect chicken anaemia virus ...

    African Journals Online (AJOL)

    To provide a rapid, easy and economical method for detecting antibodies to chicken anaemia virus (CAV) especially in large numbers of chicken sera, we established a monoclonal blocking enzyme-linked immunosorbent assay (MBE). A monoclonal antibody (MAb), 2A9, directed against the 52 kDa protein of the ...

  10. Rapid antigen detection test for respiratory syncytial virus diagnosis as a diagnostic tool,

    Directory of Open Access Journals (Sweden)

    Flávio da Silva Mesquita

    Full Text Available Abstract Objective: The aim of this study was to evaluate the QuickVue® RSV Test Kit (QUIDEL Corp, CA, USA as a screening tool for respiratory syncytial virus in children with acute respiratory disease in comparison with the indirect immunofluorescence assay as gold standard. In Brazil, rapid antigen detection tests for respiratory syncytial virus are not routinely utilized as a diagnostic tool, except for the diagnosis of dengue and influenza. Methods: The authors retrospectively analyzed 486 nasopharyngeal aspirate samples from children under age 5 with acute respiratory infection, between December 2013 and August 2014, the samples were analyzed by indirect immunofluorescence assay and QuickVue® RSV Test kit. Samples with discordant results were analyzed by real time PCR and nucleotide sequencing. Results: From 313 positive samples by immunofluorescence assays, 282 (90% were also positive by the rapid antigen detection test, two were positive only by rapid antigen detection test, 33 were positive only by immunofluorescence assays, and 171 were positive by both methods. The 35 samples with discordant results were analyzed by real time PCR; the two samples positive only by rapid antigen detection test and the five positive only by immunofluorescence assays were also positive by real time PCR. There was no relation between the negativity by QuickVue® RSV Test and viral load or specific strain. The QuickVue® RSV Test showed sensitivity of 90%, specificity of 98.8%, predictive positive value of 99.3%, and negative predictive value of 94.6%, with accuracy of 93.2% and agreement κ index of 0.85 in comparison to immunofluorescence assay. Conclusions: This study demonstrated that the QuickVue® RSV Test Kit can be effective in early detection of Respiratory syncytial virus in nasopharyngeal aspirate and is reliable for use as a diagnostic tool in pediatrics.

  11. Rapid antigen detection test for respiratory syncytial virus diagnosis as a diagnostic tool.

    Science.gov (United States)

    Mesquita, Flávio da Silva; Oliveira, Danielle Bruna Leal de; Crema, Daniela; Pinez, Célia Miranda Nunes; Colmanetti, Thaís Cristina; Thomazelli, Luciano Matsumia; Gilio, Alfredo Elias; Vieira, Sandra Elisabeth; Martinez, Marina Baquerizo; Botosso, Viviane Fongaro; Durigon, Edison Luiz

    The aim of this study was to evaluate the QuickVue® RSV Test Kit (QUIDEL Corp, CA, USA) as a screening tool for respiratory syncytial virus in children with acute respiratory disease in comparison with the indirect immunofluorescence assay as gold standard. In Brazil, rapid antigen detection tests for respiratory syncytial virus are not routinely utilized as a diagnostic tool, except for the diagnosis of dengue and influenza. The authors retrospectively analyzed 486 nasopharyngeal aspirate samples from children under age 5 with acute respiratory infection, between December 2013 and August 2014, the samples were analyzed by indirect immunofluorescence assay and QuickVue® RSV Test kit. Samples with discordant results were analyzed by real time PCR and nucleotide sequencing. From 313 positive samples by immunofluorescence assays, 282 (90%) were also positive by the rapid antigen detection test, two were positive only by rapid antigen detection test, 33 were positive only by immunofluorescence assays, and 171 were positive by both methods. The 35 samples with discordant results were analyzed by real time PCR; the two samples positive only by rapid antigen detection test and the five positive only by immunofluorescence assays were also positive by real time PCR. There was no relation between the negativity by QuickVue® RSV Test and viral load or specific strain. The QuickVue® RSV Test showed sensitivity of 90%, specificity of 98.8%, predictive positive value of 99.3%, and negative predictive value of 94.6%, with accuracy of 93.2% and agreement κ index of 0.85 in comparison to immunofluorescence assay. This study demonstrated that the QuickVue® RSV Test Kit can be effective in early detection of Respiratory syncytial virus in nasopharyngeal aspirate and is reliable for use as a diagnostic tool in pediatrics. Copyright © 2016 Sociedade Brasileira de Pediatria. Published by Elsevier Editora Ltda. All rights reserved.

  12. Viruses in Marine Animals: Discovery, Detection, and Characterization

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    Fahsbender, Elizabeth

    Diseases in marine animals are emerging at an increasing rate. Disease forecasting enabled by virus surveillance presents a proactive solution for managing emerging diseases. Broad viral surveys aid in disease forecasting by providing baseline data on viral diversity associated with various hosts, including many that are not associated with disease. However, these viruses can become pathogens due to expansion in host or geographic range, as well as when changing conditions shift the balance between commensal viruses and the host immune system. Therefore, it is extremely valuable to identify and characterize viruses present in many different hosts in a variety of environments, regardless of whether the hosts are symptomatic or not. The lack of a universal gene shared by all viruses makes virus surveillance difficult, because no single assay exists that can detect the enormous diversity of viruses. Viral metagenomics circumvents this issue by purifying viral particles directly from host tissues and sequencing the nucleic acids, allowing for virus identification. However, virus identification is only the first step, which should ideally be followed by complete sequencing of the viral genome to identify genes of interest and develop assays to reveal viral prevalence, tropism, ecology, and pathogenicity. This dissertation focuses on the discovery of novel viruses in marine animals, characterization of complete viral genomes, and the development of subsequent diagnostic assays for further analysis of virus ecology. First, viral metagenomics was used to explore the viruses present in the healthy Weddell seal (Leptonychotes weddellii) population in Antarctica, which led to the discovery of highly prevalent small, circular single-stranded DNA (ssDNA) viruses. The lack of knowledge regarding the viruses of Antarctic wildlife warrants this study to determine baseline viral communities in healthy animals that can be used to survey changes over time. From the healthy Weddell

  13. Using Fluorescent Viruses for Detecting Bacteria in Water

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    Tabacco, Mary Beth; Qian, Xiaohua; Russo, Jaimie A.

    2009-01-01

    A method of detecting water-borne pathogenic bacteria is based partly on established molecular-recognition and fluorescent-labeling concepts, according to which bacteria of a species of interest are labeled with fluorescent reporter molecules and the bacteria can then be detected by fluorescence spectroscopy. The novelty of the present method lies in the use of bacteriophages (viruses that infect bacteria) to deliver the fluorescent reporter molecules to the bacteria of the species of interest.

  14. Genotypic Characterization of Herpes Simplex Virus Type 1 Isolates in Immunocompromised Patients in Rio de Janeiro, Brazil.

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    Amanda Perse da Silva

    Full Text Available Herpes simplex virus type 1 (HSV-1 is a prevalent human pathogen that causes a variety of diseases, including an increased risk of developing more severe disease in HIV-infected individuals. In Brazil, there is no information about the molecular epidemiology of HSV-1 infection, especially in HIV-infected individuals. The aim of this study was to perform the genotypic characterization of HSV-1 among HIV-infected patients. A total of 214 serum samples from HIV-positive patients without HSV infection symptoms were enrolled in one of two reference hospitals for HIV infection managing in Rio de Janeiro. The gG and gI genes were analyzed by restriction fragment length polymorphism (RFLP and full nucleotide sequencing of the US8 (1601 bp, UL44 (1996 bp, and UL23 (1244 bp regions was performed. A total of 38.3% (82/214 and 32.7% (70/214 of the serum samples tested positive for gG and gI genes, respectively. RFLP analysis classified the HSV-1 as belonging to genotype A. Phylogenetic analysis of the Brazilian samples for the US8, UL44, and UL23 regions demonstrated that the nucleotide identity between Brazilian samples was higher than 97% for all genes. No acyclovir mutation was detected in the patients. The shedding of HSV in the serum samples from HIV-positive patients who were asymptomatic for HSV infection was detected in this work. This is the first report of molecular characterization of HSV-1 in Brazilian samples since there is no previous data available in the literature concerning the genotypic classification and stable distribution of Brazilian strains of HSV-1 in Rio de Janeiro, Brazil.

  15. Genotypic Characterization of Herpes Simplex Virus Type 1 Isolates in Immunocompromised Patients in Rio de Janeiro, Brazil

    Science.gov (United States)

    Perse da Silva, Amanda; Lopes, Amanda de Oliveira; Vieira, Yasmine Rangel; de Almeida, Adilson José; Sion, Fernando Samuel; Grinsztejn, Beatriz; Wagner, Sandra; de Paula, Vanessa Salete

    2015-01-01

    Herpes simplex virus type 1 (HSV-1) is a prevalent human pathogen that causes a variety of diseases, including an increased risk of developing more severe disease in HIV-infected individuals. In Brazil, there is no information about the molecular epidemiology of HSV-1 infection, especially in HIV-infected individuals. The aim of this study was to perform the genotypic characterization of HSV-1 among HIV-infected patients. A total of 214 serum samples from HIV-positive patients without HSV infection symptoms were enrolled in one of two reference hospitals for HIV infection managing in Rio de Janeiro. The gG and gI genes were analyzed by restriction fragment length polymorphism (RFLP) and full nucleotide sequencing of the US8 (1601 bp), UL44 (1996 bp), and UL23 (1244 bp) regions was performed. A total of 38.3% (82/214) and 32.7% (70/214) of the serum samples tested positive for gG and gI genes, respectively. RFLP analysis classified the HSV-1 as belonging to genotype A. Phylogenetic analysis of the Brazilian samples for the US8, UL44, and UL23 regions demonstrated that the nucleotide identity between Brazilian samples was higher than 97% for all genes. No acyclovir mutation was detected in the patients. The shedding of HSV in the serum samples from HIV-positive patients who were asymptomatic for HSV infection was detected in this work. This is the first report of molecular characterization of HSV-1 in Brazilian samples since there is no previous data available in the literature concerning the genotypic classification and stable distribution of Brazilian strains of HSV-1 in Rio de Janeiro, Brazil. PMID:26407292

  16. Prevalence of serological markers of hepatitis B virus in pregnant women from Paraná State, Brazil

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    D.A. Bertolini

    2006-08-01

    Full Text Available The prevalence of hepatitis B virus (HBV in Brazil increases from South to North but moderate to elevated prevalence has been detected in the Southwest of Paraná State. The prevalence of serological markers of HBV was evaluated in 3188 pregnant women from different counties in Paraná State and relevant epidemiological features were described. The prevalence of HBV markers in pregnant women for the state as a whole was 18.5% (95% CI = 17.2-19.9, ranging from 7.2% in Curitiba to 38.5% in Francisco Beltrão. The endemicity of HBV marker prevalence in pregnant women was intermediate in Cascavel, Foz do Iguaçu, and Francisco Beltrão, and low in Curitiba, Londrina, Maringá, and Paranaguá. Multiple logistic regression showed that HBV marker prevalence increased with age, was higher among black women, among women of Italian and German descent, and among women who had family members in neighboring Rio Grande do Sul State. Univariate analysis showed that HBV marker prevalence was also higher among women with no education or only primary education, with a lower family income and whose families originated from the South Region of Brazil. Pregnant women not having positive HBV markers (anti-HBc, HBsAg or anti-HBs detected by ELISA corresponded to 73.7% of the population studied, implying that HBV vaccination needs to be reinforced in Paraná State. The highest prevalence was found in three counties that received the largest number of families from Santa Catarina and Rio Grande do Sul, where most immigrants were of German or Italian ascendance. This finding probably indicates that immigrants that came to this area brought HBV infection to Southwestern Paraná State.

  17. Hepatitis B Virus Genotype D Isolates Circulating in Chapeco, Southern Brazil, Originate from Italy.

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    Carolina Souza Gusatti

    Full Text Available Hepatitis B virus genotype A1 (HBV/A1, of African origin, is the most prevalent genotype in Brazil, while HBV/F predominates in the other South American countries. However, HBV/D is the most common in the three states of southern Brazil, where 'islands' of elevated prevalence, as Chapecó and other cities, have been described. In this study, 202 HBV chronic carriers attending in 2013 the viral hepatitis ambulatory of Chapecó, were investigated. In comparison with previous studies performed in the same ambulatory, a rapid aging of the HBV infected population was observed (mean age of the newly diagnosed patients increasing from 29.9 ± 10.3 years in 1996 to 44.4 ± 13.3 years in 2013, probably due to a singular vaccination schedule at Chapecó that included not only children but also adolescents. Phylogenetic and BLAST analyses (S region classified 91 HBV isolates into genotypes A (n = 3 and D (n = 88. The majority of HBV/D isolates were closely related to D3 sequences. To understand the reasons for the absence or near absence of genotypes A and F, and how HBV/D was introduced in the south of Brazil, HBV/D infected patients were inquired about their genealogical and geographical origins. Forty-three (52% patients have their four grandparents of Italian origin, vs. seven (8% who have their four grandparents of Brazilian origin. At all, 65 out of 83 (78% patients had at least one grandparent originating from Italy. Taking into consideration the fact that Italy is one of the few countries where subgenotype D3 is predominant, the results strongly suggested that HBV/D was introduced in Brazil through Italian immigration which culminated between 1870 and 1920.

  18. Simultaneous Detection of Barley Virus Diseases in Korea

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    Bong-Choon Lee

    2017-12-01

    Full Text Available Barley mild mosaic virus (BaMMV, Barley yellow mosaic virus (BaYMV and Barley yellow dwarf virus (BYDV have been identified as an important causative agents for an economically important disease of winter barley in Korea. In this study, a multiplex reverse transcription polymerase chain reaction (mRT-PCR method was used for the simultaneous detection. Three sets of virus-specific primers targeted to the capsid protein coding genes of BaMMV, BaYMV and BYDV were used to amplify fragments that were 594 bp, 461 bp, and 290 bp, respectively. Several sets of primers for each target virus were evaluated for their sensitivity and specificity by multiplex RT-PCR. The optimum primer concentrations and RT-PCR conditions were determined for the multiplex RT-PCR. The mRT-PCR assay was found to be a better and rapid virus diagnostic tool of specific barley diseases and potential for investigating the epidemiology of these viral diseases.

  19. Virus detection using Viro-Adembeads, a rapid capture system for viruses, and plaque assay in intentionally virus-contaminated beverages.

    Science.gov (United States)

    Hatano, Ben; Kojima, Asato; Sata, Tetsutaro; Katano, Harutaka

    2010-01-01

    Intentional contamination of beverages with microbes is one type of bioterrorist threat. While bacteria and fungus can be easily collected by a centrifuge, viruses are difficult to collect from virus-contaminated beverages. In this study, we demonstrated that Viro-Adembeads, a rapid-capture system for viruses using anionic polymer-coated magnetic beads, collected viruses from beverages contaminated intentionally with vaccinia virus and human herpesvirus 8. Real-time PCR showed that the recovery rates of the contaminated viruses in green tea and orange juice were lower than those in milk and water. Plaque assay showed that green tea and orange juice cut the efficiency of vaccinia virus infection in CV-1 cells. These results suggest that the efficiency of virus detection depends on the kind of beverage being tested. Viro-Adembeads would be a useful tool for detecting viruses rapidly in virus-contaminated beverages used in a bioterrorist attack.

  20. Rapid Spread of Zika Virus in The Americas - Implications for Public Health Preparedness for Mass Gatherings at the 2016 Brazil Olympic Games

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    Eskild Petersen

    2016-03-01

    Full Text Available Mass gatherings at major international sporting events put millions of international travelers and local host-country residents at risk of acquiring infectious diseases, including locally endemic infectious diseases. The mosquito-borne Zika virus (ZIKV has recently aroused global attention due to its rapid spread since its first detection in May 2015 in Brazil to 22 other countries and other territories in the Americas. The ZIKV outbreak in Brazil, has also been associated with a significant rise in the number of babies born with microcephaly and neurological disorders, and has been declared a ‘Global Emergency by the World Health Organization. This explosive spread of ZIKV in Brazil poses challenges for public health preparedness and surveillance for the Olympics and Paralympics which are due to be held in Rio De Janeiro in August, 2016. We review the epidemiology and clinical features of the current ZIKV outbreak in Brazil, highlight knowledge gaps, and review the public health implications of the current ZIKV outbreak in the Americas. We highlight the urgent need for a coordinated collaborative response for prevention and spread of infectious diseases with epidemic potential at mass gatherings events.

  1. Detecting emerging transmissibility of avian influenza virus in human households.

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    Michiel van Boven

    2007-07-01

    Full Text Available Accumulating infections of highly pathogenic H5N1 avian influenza in humans underlines the need to track the ability of these viruses to spread among humans. A human-transmissible avian influenza virus is expected to cause clusters of infections in humans living in close contact. Therefore, epidemiological analysis of infection clusters in human households is of key importance. Infection clusters may arise from transmission events from (i the animal reservoir, (ii humans who were infected by animals (primary human-to-human transmission, or (iii humans who were infected by humans (secondary human-to-human transmission. Here we propose a method of analysing household infection data to detect changes in the transmissibility of avian influenza viruses in humans at an early stage. The method is applied to an outbreak of H7N7 avian influenza virus in The Netherlands that was the cause of more than 30 human-to-human transmission events. The analyses indicate that secondary human-to-human transmission is plausible for the Dutch household infection data. Based on the estimates of the within-household transmission parameters, we evaluate the effectiveness of antiviral prophylaxis, and conclude that it is unlikely that all household infections can be prevented with current antiviral drugs. We discuss the applicability of our method for the detection of emerging human-to-human transmission of avian influenza viruses in particular, and for the analysis of within-household infection data in general.

  2. Avian infectious bronchitis virus in Brazil: a highly complex virus meets a highly susceptible host population

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    PE Brandão

    2010-06-01

    Full Text Available Infectious bronchitis (IB is a highly aggressive disease for poultry in terms of symptoms and economic losses, and the control of this disease is difficult if flocks are not protected against type-specific challenges by the Avian infectious bronchitis virus (IBV. This article summarizes data presented by the author at the Workshop on Infectious Bronchitis 2009 on IB and IBV, including future developments on the field.

  3. Detection of arboviruses of public health interest in free-living New World primates (Sapajus spp.; Alouatta caraya captured in Mato Grosso do Sul, Brazil

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    Paulo Mira Batista

    2013-12-01

    Full Text Available Introduction A sero-epidemiological survey was undertaken to detect the circulation of arboviruses in free-living non-human primates. Methods Blood samples were obtained from 16 non-human primates (13 Sapajus spp. and three Alouatta caraya that were captured using terrestrial traps and anesthetic darts in woodland regions in the municipalities of Campo Grande, Aquidauana, Jardim, Miranda and Corumbá in the State of Mato Grosso do Sul, Brazil. The samples were sent to the Instituto Evandro Chagas (IEC in Ananindeua, Pará, Brazil, to detect antibodies against 19 species of arboviruses using a hemagglutination inhibition test (HI. Results Of the 16 primates investigated in the present study, five (31.2% were serologically positive for an arbovirus. Of these five, two (12.5% exhibited antibodies to the Flavivirus genus, one (6.2% exhibited a monotypic reaction to Cacipacoré virus, one (6.2% was associated with Mayaro virus, and one (6.2% was positive for Oropouche virus. Conclusions Based on the positive serology observed in the present study, it was possible to conclude that arboviruses circulate among free-living primates. The viruses in the areas studied might have been introduced by infected humans or by primates from endemic or enzootic areas. Studies of this nature, as well as efficient and continuous surveillance programs, are needed to monitor viral activities in endemic and enzootic regions.

  4. Rapid detection of anti-Vaccinia virus neutralizing antibodies

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    Lichtfuss Gregor F

    2011-03-01

    Full Text Available Abstract Increasing infections with Monkeypox and Cowpox viruses pose a continuous and growing threat to human health. The standard method for detecting poxvirus neutralizing antibodies is the plaque-reduction neutralization test that is specific but also time-consuming and laborious. Therefore, a rapid and reliable method was developed to determine neutralizing antibody titers within twelve hours. The new assay measures viral mRNA transcription as a marker for actively replicating virus after incomplete neutralization using real-time PCR.

  5. Association between Zika virus infection and microcephaly in Brazil, January to May, 2016: preliminary report of a case-control study.

    Science.gov (United States)

    de Araújo, Thalia Velho Barreto; Rodrigues, Laura Cunha; de Alencar Ximenes, Ricardo Arraes; de Barros Miranda-Filho, Demócrito; Montarroyos, Ulisses Ramos; de Melo, Ana Paula Lopes; Valongueiro, Sandra; de Albuquerque, Maria de Fátima Pessoa Militão; Souza, Wayner Vieira; Braga, Cynthia; Filho, Sinval Pinto Brandão; Cordeiro, Marli Tenório; Vazquez, Enrique; Di Cavalcanti Souza Cruz, Danielle; Henriques, Cláudio Maierovitch Pessanha; Bezerra, Luciana Caroline Albuquerque; da Silva Castanha, Priscila Mayrelle; Dhalia, Rafael; Marques-Júnior, Ernesto Torres Azevedo; Martelli, Celina Maria Turchi

    2016-12-01

    The microcephaly epidemic, which started in Brazil in 2015, was declared a Public Health Emergency of International Concern by WHO in 2016. We report the preliminary results of a case-control study investigating the association between microcephaly and Zika virus infection during pregnancy. We did this case-control study in eight public hospitals in Recife, Brazil. Cases were neonates with microcephaly. Two controls (neonates without microcephaly), matched by expected date of delivery and area of residence, were selected for each case. Serum samples of cases and controls and cerebrospinal fluid samples of cases were tested for Zika virus-specific IgM and by quantitative RT-PCR. Laboratory-confirmed Zika virus infection during pregnancy was defined as detection of Zika virus-specific IgM or a positive RT-PCR result in neonates. Maternal serum samples were tested by plaque reduction neutralisation assay for Zika virus and dengue virus. We estimated crude odds ratios (ORs) and 95% CIs using a median unbiased estimator for binary data in an unconditional logistic regression model. We estimated ORs separately for cases with and without radiological evidence of brain abnormalities. Between Jan 15, 2016, and May 2, 2016, we prospectively recruited 32 cases and 62 controls. 24 (80%) of 30 mothers of cases had Zika virus infection compared with 39 (64%) of 61 mothers of controls (p=0·12). 13 (41%) of 32 cases and none of 62 controls had laboratory-confirmed Zika virus infection; crude overall OR 55·5 (95% CI 8·6-∞); OR 113·3 (95% CI 14·5-∞) for seven cases with brain abnormalities; and OR 24·7 (95% CI 2·9-∞) for four cases without brain abnormalities. Our data suggest that the microcephaly epidemic is a result of congenital Zika virus infection. We await further data from this ongoing study to assess other potential risk factors and to confirm the strength of association in a larger sample size. Brazilian Ministry of Health, Pan American Health Organization

  6. Ocular Findings in Infants With Microcephaly Associated With Presumed Zika Virus Congenital Infection in Salvador, Brazil.

    Science.gov (United States)

    de Paula Freitas, Bruno; de Oliveira Dias, João Rafael; Prazeres, Juliana; Sacramento, Gielson Almeida; Ko, Albert Icksang; Maia, Maurício; Belfort, Rubens

    2016-02-09

    The Zika virus (ZIKV) has rapidly reached epidemic proportions, especially in northeastern Brazil, and has rapidly spread to other parts of the Americas. A recent increase in the prevalence of microcephaly in newborn infants and vision-threatening findings in these infants is likely associated with the rapid spread of ZIKV. To evaluate the ocular findings in infants with microcephaly associated with presumed intrauterine ZIKV infection in Salvador, Bahia, Brazil. Case series at a tertiary hospital. Twenty-nine infants with microcephaly (defined by a cephalic circumference of ≤32 cm) with a presumed diagnosis of congenital ZIKV were recruited through an active search and referrals from other hospitals and health unities. The study was conducted between December 1 and December 21, 2015. All infants and mothers underwent systemic and ophthalmic examinations from December 1 through December 21, 2015, in the Roberto Santos General Hospital, Salvador, Brazil. Anterior segment and retinal, choroidal, and optic nerve abnormalities were documented using a wide-field digital imaging system. The differential diagnosis included toxoplasmosis, rubella, cytomegalovirus, herpes simplex virus, syphilis, and human immunodeficiency virus, which were ruled out through serologic and clinical examinations. Ocular abnormalities associated with ZIKV. Twenty-three of 29 mothers (79.3%) reported suspected ZIKV infection signs and symptoms during pregnancy, 18 in the first trimester, 4 in the second trimester, and 1 in the third trimester. Of the 29 infants (58 eyes) examined (18 [62.1%] female), ocular abnormalities were present in 17 eyes (29.3%) of 10 children (34.5%). Bilateral findings were found in 7 of 10 patients presenting with ocular lesions, the most common of which were focal pigment mottling of the retina and chorioretinal atrophy in 11 of the 17 eyes with abnormalities (64.7%), followed by optic nerve abnormalities in 8 eyes (47.1%), bilateral iris coloboma in 1 patient (2

  7. [Simultaneous detection of respiratory viruses and influenza A virus subtypes using multiplex PCR].

    Science.gov (United States)

    Ciçek, Candan; Bayram, Nuri; Anıl, Murat; Gülen, Figen; Pullukçu, Hüsnü; Saz, Eylem Ulaş; Telli, Canan; Cok, Gürsel

    2014-10-01

    This study was conducted to investigate the respiratory viruses and subtyping of influenza A virus when positive by multiplex PCR in patients with flu-like symptoms, after the pandemic caused by influenza A (H1N1)pdm09. Nasopharyngeal swab samples collected from 700 patients (313 female, 387 male; age range: 24 days-94 yrs, median age: 1 yr) between December 2010 - January 2013 with flu-like symptoms including fever, headache, sore throat, rhinitis, cough, myalgia as defined by the World Health Organization were included in the study. Nucleic acid extractions (Viral DNA/RNA Extraction Kit, iNtRON, South Korea) and cDNA synthesis (RevertAid First Strand cDNA Synthesis Kits, Fermentas, USA) were performed according to the manufacturer's protocol. Multiplex amplification of nucleic acids was performed using DPO (dual priming oligonucleotide) primers and RV5 ACE Screening Kit (Seegene, South Korea) in terms of the presence of influenza A (INF-A) virus, influenza B (INF-B) virus, respiratory syncytial virus (RSV), and the other respiratory viruses. PCR products were detected by automated polyacrylamide gel electrophoresis using Screen Tape multiple detection system. Specimens which were positive for viral nucleic acids have been further studied by using specific DPO primers, FluA ACE Subtyping and RV15 Screening (Seegene, South Korea) kits. Four INF-A virus subtypes [human H1 (hH1), human H3 (hH3), swine H1 (sH1), avian H5 (aH5)] and 11 other respiratory viruses [Adenovirus, parainfluenza virus (PIV) types 1-4, human bocavirus (HBoV), human metapneumovirus (HMPV), rhinovirus types A and B, human coronaviruses (HCoV) OC43, 229E/NL63] were investigated with those tests. In the study, 53.6% (375/700) of the patients were found to be infected with at least one virus and multiple respiratory virus infections were detected in 15.7% (59/375) of the positive cases, which were mostly (49/59, 83%) in pediatric patients. RSV and rhinovirus coinfections were the most prevalent (18

  8. Prevalence of Epstein-Barr virus antibodies in healthy children and adolescents in Vitória, State of Espírito Santo, Brazil

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    Figueira-Silva Cecília M.

    2004-01-01

    Full Text Available The prevalence and age distribution of Epstein-Barr virus infection varies in different populations and there is little information about the epidemiology of this infection in Brazil. We studied the prevalence of EBV antibodies in a sample of 283 children and adolescents between 1 and 21 years old. The sample was taken from two neighborhoods in Vitória (capital city of Espirito Santo, Brazil. The São Pedro (SP neighborhood represented an area with lower socioeconomic status and the Praias (P neighborhood represented an area with higher SES. Anti-VCA (Virus Capsid Antigen antibodies were detected by ELISA and anti-EBNA (Epstein-Barr Nuclear Antigen antibodies were detected by an anti-complement immunofluorescence method, both using commercial kits. The results showed an overall prevalence rates of anti-VCA and anti-EBNA of 71% and 54% respectively. The prevalence for both anti-EBV antibodies was higher and probably the infection occurred earlier in the SP neighborhood. Among the various socioeconomic factors studied only low family income and maternal education level were significantly correlated with a higher frequency of positive serology for anti-VCA. These results demonstrate that there is a high prevalence of EBV antibodies in children and adolescents living in Vitória, that occurs more frequently at a younger age in children from families with low socioeconomic status. In addition, the results demonstrate an intermediate age distribution pattern between those reported in developed and underdeveloped countries.

  9. Highly Specific Detection of Five Exotic Quarantine Plant Viruses using RT-PCR

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    Hoseong Choi; Won Kyong Cho; Jisuk Yu; Jong-Seung Lee; Kook-Hyung Kim

    2013-01-01

    To detect five plant viruses (Beet black scorch virus, Beet necrotic yellow vein virus, Eggplant mottled dwarf virus, Pelargonium zonate spot virus, and Rice yellow mottle virus) for quarantine purposes, we designed 15 RT-PCR primer sets. Primer design was based on the nucleotide sequence of the coat protein gene, which is highly conserved within species. All but one primer set successfully amplified the targets, and gradient PCRs indicated that the optimal temperature for the 14 useful prime...

  10. Hepatitis B virus genotypes circulating in Brazil: molecular characterization of genotype F isolates.

    Science.gov (United States)

    Mello, Francisco C A; Souto, Francisco J D; Nabuco, Leticia C; Villela-Nogueira, Cristiane A; Coelho, Henrique Sergio M; Franz, Helena Cristina F; Saraiva, Joao Carlos P; Virgolino, Helaine A; Motta-Castro, Ana Rita C; Melo, Mabel M M; Martins, Regina M B; Gomes, Selma A

    2007-11-23

    Hepatitis B virus (HBV) isolates have been classified in eight genotypes, A to H, which exhibit distinct geographical distributions. Genotypes A, D and F are predominant in Brazil, a country formed by a miscegenated population, where the proportion of individuals from Caucasian, Amerindian and African origins varies by region. Genotype F, which is the most divergent, is considered indigenous to the Americas. A systematic molecular characterization of HBV isolates from different parts of the world would be invaluable in establishing HBV evolutionary origins and dispersion patterns. A large-scale study is needed to map the region-by-region distribution of the HBV genotypes in Brazil. Genotyping by PCR-RFLP of 303 HBV isolates from HBsAg-positive blood donors showed that at least two of the three genotypes, A, D, and F, co-circulate in each of the five geographic regions of Brazil. No other genotypes were identified. Overall, genotype A was most prevalent (48.5%), and most of these isolates were classified as subgenotype A1 (138/153; 90.2%). Genotype D was the most common genotype in the South (84.2%) and Central (47.6%) regions. The prevalence of genotype F was low (13%) countrywide. Nucleotide sequencing of the S gene and a phylogenetic analysis of 32 HBV genotype F isolates showed that a great majority (28/32; 87.5%) belonged to subgenotype F2, cluster II. The deduced serotype of 31 of 32 F isolates was adw4. The remaining isolate showed a leucine-to-isoleucine substitution at position 127. The presence of genotypes A, D and F, and the absence of other genotypes in a large cohort of HBV infected individuals may reflect the ethnic origins of the Brazilian population. The high prevalence of isolates from subgenotype A1 (of African origin) indicates that the African influx during the colonial slavery period had a major impact on the circulation of HBV genotype A currently found in Brazil. Although most genotype F isolates belonged to cluster II, the presence of some

  11. Hepatitis B virus genotypes circulating in Brazil: molecular characterization of genotype F isolates

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    Virgolino Helaine A

    2007-11-01

    Full Text Available Abstract Background Hepatitis B virus (HBV isolates have been classified in eight genotypes, A to H, which exhibit distinct geographical distributions. Genotypes A, D and F are predominant in Brazil, a country formed by a miscegenated population, where the proportion of individuals from Caucasian, Amerindian and African origins varies by region. Genotype F, which is the most divergent, is considered indigenous to the Americas. A systematic molecular characterization of HBV isolates from different parts of the world would be invaluable in establishing HBV evolutionary origins and dispersion patterns. A large-scale study is needed to map the region-by-region distribution of the HBV genotypes in Brazil. Results Genotyping by PCR-RFLP of 303 HBV isolates from HBsAg-positive blood donors showed that at least two of the three genotypes, A, D, and F, co-circulate in each of the five geographic regions of Brazil. No other genotypes were identified. Overall, genotype A was most prevalent (48.5%, and most of these isolates were classified as subgenotype A1 (138/153; 90.2%. Genotype D was the most common genotype in the South (84.2% and Central (47.6% regions. The prevalence of genotype F was low (13% countrywide. Nucleotide sequencing of the S gene and a phylogenetic analysis of 32 HBV genotype F isolates showed that a great majority (28/32; 87.5% belonged to subgenotype F2, cluster II. The deduced serotype of 31 of 32 F isolates was adw4. The remaining isolate showed a leucine-to-isoleucine substitution at position 127. Conclusion The presence of genotypes A, D and F, and the absence of other genotypes in a large cohort of HBV infected individuals may reflect the ethnic origins of the Brazilian population. The high prevalence of isolates from subgenotype A1 (of African origin indicates that the African influx during the colonial slavery period had a major impact on the circulation of HBV genotype A currently found in Brazil. Although most genotype F

  12. A multiplex RT-PCR for rapid and simultaneous detection of viruses and viroids in chrysanthemum.

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    Song, A; You, Y; Chen, F; Li, P; Jiang, J; Chen, S

    2013-01-01

    Chrysanthemum plants are subject to serious virus diseases, so detection and identification of virus pathogens is important to prevent the virus spread. A reliable one-step multiplex RT-PCR was developed to simultaneously detect two viruses and two viriods: chrysanthemum virus B, tomato Aspermy virus, chrysanthemum stunt viroid and chrysanthemum chlorotic mottle viroid. In addition, we investigated the detection limit and the efficiency of single and multiplex RT-PCR assays. The results showed that the multiplex RT-PCR assay proved to be as sensitive as the single one. In conclusion, this technique is potentially useful in routine diagnosis of chrysanthemum viruses and viroids. The multiplex RT-PCR assay described in this study is the first report of simultaneous detection of virus and viroid in chrysanthemum, which provides a fast, convenient, cost-saving way to detect the virus and viroid mixed infections in plants. © 2012 The Society for Applied Microbiology.

  13. A method for simultaneous detection and identification of Brazilian dog- and vampire bat-related rabies virus by reverse transcription loop-mediated isothermal amplification assay.

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    Saitou, Yasumasa; Kobayashi, Yuki; Hirano, Shinji; Mochizuki, Nobuyuki; Itou, Takuya; Ito, Fumio H; Sakai, Takeo

    2010-09-01

    At present, the sporadic occurrence of human rabies in Brazil can be attributed primarily to dog- and vampire bat-related rabies viruses. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) was employed as a simultaneous detection method for both rabies field variants within 60 min. Vampire bat-related rabies viruses could be distinguished from dog variants by digesting amplicons of the RT-LAMP reaction using the restriction enzyme AlwI. Amplification and digestion could both be completed within 120 min after RNA extraction. In addition, the RT-LAMP assay also detected rabies virus in isolates from Brazilian frugivorous bats and Ugandan dog, bovine and goat samples. In contrast, there were false negative results from several Brazilian insectivorous bats and all of Chinese dog, pig, and bovine samples using the RT-LAMP assay. This study showed that the RT-LAMP assay is effective for the rapid detection of rabies virus isolates from the primary reservoir in Brazil. Further improvements are necessary so that the RT-LAMP assay can be employed for the universal detection of genetic variants of rabies virus in the field. Copyright 2010 Elsevier B.V. All rights reserved.

  14. High Zika Virus Seroprevalence in Salvador, Northeastern Brazil Limits the Potential for Further Outbreaks

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    Eduardo Martins Netto

    2017-11-01

    Full Text Available During 2015 to 2016, Brazil reported more Zika virus (ZIKV cases than any other country, yet population exposure remains unknown. Serological studies of ZIKV are hampered by cross-reactive immune responses against heterologous viruses. We conducted serosurveys for ZIKV, dengue virus (DENV, and Chikungunya virus (CHIKV in 633 individuals prospectively sampled during 2015 to 2016, including microcephaly and non-microcephaly pregnancies, HIV-infected patients, tuberculosis patients, and university staff in Salvador in northeastern Brazil using enzyme-linked immunosorbent assays (ELISAs and plaque reduction neutralization tests. Sera sampled retrospectively during 2013 to 2015 from 277 HIV-infected patients were used to assess the spread of ZIKV over time. Individuals were georeferenced, and sociodemographic indicators were compared between ZIKV-positive and -negative areas and areas with and without microcephaly cases. Epidemiological key parameters were modeled in a Bayesian framework. ZIKV seroprevalence increased rapidly during 2015 to 2016, reaching 63.3% by 2016 (95% confidence interval [CI], 59.4 to 66.8%, comparable to the seroprevalence of DENV (75.7%; CI, 69.4 to 81.1% and higher than that of CHIKV (7.4%; CI, 5.6 to 9.8%. Of 19 microcephaly pregnancies, 94.7% showed ZIKV IgG antibodies, compared to 69.3% of 257 non-microcephaly pregnancies (P = 0.017. Analyses of sociodemographic data revealed a higher ZIKV burden in low socioeconomic status (SES areas. High seroprevalence, combined with case data dynamics allowed estimates of the basic reproduction number R0 of 2.1 (CI, 1.8 to 2.5 at the onset of the outbreak and an effective reproductive number Reff of <1 in subsequent years. Our data corroborate ZIKV-associated congenital disease and an association of low SES and ZIKV infection and suggest that population immunity caused cessation of the outbreak. Similar studies from other areas will be required to determine the fate of the American

  15. Macroarray detection of grapevine leafroll-associated viruses.

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    Thompson, Jeremy R; Fuchs, Marc; Fischer, Kael F; Perry, Keith L

    2012-08-01

    Grapevine leafroll-associated viruses (GLRaVs) are an emerging group of viruses that represent a significant threat to the global productivity and sustainability of the grapevine industry. Their control is achieved through the identification and elimination of infected vines, and the use of planting material derived from virus-tested, certified stocks. As such, much effort has been invested in developing reliable molecular diagnostic techniques. In this work, we report the development of a macroarray assay for the detection of the principal GLRaVs. In total 314 70-mer oligonucleotides specific to GLRaV-1, -2, -3, -4, -7, and GLRaV-4 strains 5, 6, 9 and Pr were spotted onto a 11×7cm nylon membrane. Thirty-four grapevine samples from various origins were tested by the macroarray, RT-PCR and ELISA. Thirty were positive for virus infection using RT-PCR, 28 by ELISA and 25 by the macroarray. Mixed infections were identified by macroarray in two samples and confirmed by RT-PCR or ELISA. There were a few discrepancies between methods that were most likely due to differences in the sensitivity of detection, and in the case of the macroarray, limitations in the sequence data available for certain virus species in the design of the oligonucleotides. This work demonstrates the successful application of macroarray methodology using randomly primed and sequence-nonspecific amplified cDNAs derived from grapevine total RNA extracts, and provides a proof-of-principal for unbiased multiplex detection using a single robust platform. Published by Elsevier B.V.

  16. Characterization of rabies virus isolated from a colony of Eptesicus furinalis bats in Brazil

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    Marilene Fernandes de Almeida

    2011-02-01

    Full Text Available Some bat species have adapted to the expanding human population by acquiring the ability to roost in urban buildings, increasing the exposure risk for people and domestic animals, and consequently, the likelihood of transmitting rabies. Three dead bats were found in the yard of a house in an urban area of Jundiaí city in the state of São Paulo in southeast Brazil. Two of the three bats tested positive for rabies, using Fluorescent Antibody and Mouse Inoculation techniques. A large colony of Eptesicus furinalis was found in the house's attic, and of the 119 bats captured, four more tested positive for rabies. The objectives of this study were to report the rabies diagnosis, characterize the isolated virus antigenically and genetically, and study the epidemiology of the colony.

  17. Avian pox virus infection in a common barn owl (Tyto alba in southern Brazil

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    Gilberto D. Vargas

    2011-07-01

    Full Text Available A young common barn owl (Tyto alba was referred to the Núcleo de Reabilitação da Fauna Silvestre (Nurfs, Federal University of Pelotas (UFPel, after been found in a barn of a brick factory in the urban area of Pelotas, Rio Grande do Sul, Brazil. The bird was apathic, weak and with crusty lesions in the featherless areas (eyes, beak, legs, and died soon after arrival at Nurfs. Necropsy and histopathological examination of the lesions were carried out. The hyperplasia and hypertrophy of the cutaneous lesions, several eosinophilic intracyto-plasmic inclusion bodies in epithelial cells (Bollinger bodies, as well as particles characteristic of poxvirus, observed by electronic microscopy, confirmed the infection by avian poxvirus, what highlights the importance of Tyto alba as carrier of the virus in the wild.

  18. Effects of bovine leukemia virus infection on crossbred and purebred dairy cattle productive performance in Brazil

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    Daniela Souza Rajão

    2014-02-01

    Full Text Available The aim of this study was to evaluate the effects of bovine leukemia virus (BLV infection on productive performance of dairy cattle in Brazil. A total of 158 blood samples from lactating adult cows, purebred Holstein and crossbred Holstein X Zebu, were analyzed by Agar Gel Immunodifusion Test (AGID and leukogram. According to AGID and leukogram results, animals were grouped into three categories: seronegative, seropositive without persistent lymphocytosis, and seropositive with persistent lymphocytosis. Milk production data were compared between groups, according to breed. BLV infected females showed lower milk yield than uninfected ones, both purebred and crossbred ones. There was no difference between milk yield of seropositive cows with or without persistent lymphocytosis. These results indicate an association between BLV infection and reduction of milk production, and this study is the first one to show these effects in crossbred Holstein X Zebu cows.

  19. Epstein-Barr virus infection and gastric carcinoma in São Paulo State, Brazil

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    L.F. Lopes

    2004-11-01

    Full Text Available Epstein-Barr virus (EBV is a ubiquitous herpesvirus, and most people have serological evidence of previous viral infection at adult age. EBV is associated with infectious mononucleosis and human cancers, including some lymphomas and gastric carcinomas. Although EBV was first reported in lymphoepithelioma-like gastric carcinoma, the virus was also found in conventional adenocarcinomas. In the present study, 53 gastric carcinomas diagnosed in São Paulo State, Brazil, were evaluated for EBV infection by non-isotopic in situ hybridization with a biotinylated probe (Biotin-AGACACCGTCCTCACCACCC GGGACTTGTA directed to the viral transcript EBER-I, which is actively expressed in EBV latently infected cells. EBV infection was found in 6 of 53 (11.32% gastric carcinomas, mostly from male patients (66.7%, with a mean age of 59 years old. Most EBV-positive tumors were in gastric antrum. Two EBV-positive tumors (33.3% were conventional adenocarcinomas, whereas four (66.7% were classified as lymphoepithelioma-like carcinomas. EBV infection in gastric carcinomas was reported elsewhere in frequencies that range from 5.6% (Korea up to 18% (Germany. In Brazil, a previous work found EBV infection in 4 of 80 (5% gastric carcinomas, whereas another study found 4.7 and 11.2% of EBV-positive gastric carcinomas of Brazilians of Japanese origin or not, respectively. In the present study, the frequency of EBV-positive gastric carcinomas is similar to that reported in other series, and the clinicopathologic characteristics of these EBV-positive tumors are in agreement with the data in the literature.

  20. Hepatitis Delta virus genotype 8 infection in Northeast Brazil: inheritance from African slaves?

    Science.gov (United States)

    Barros, L M F; Gomes-Gouvêa, M S; Pinho, J R R; Alvarado-Mora, M V; Dos Santos, A; Mendes-Corrêa, M C J; Caldas, A J M; Sousa, M T; Santos, M D C; Ferreira, A S P

    2011-09-01

    Hepatitis Delta virus (HDV) is endemic worldwide, but its prevalence varies in different geographical areas. While in the Brazilian Amazon, HDV is known to be endemic and to represent a significant public health problem, few studies have assessed its prevalence in other regions in the country. This study evaluated the seroprevalence of HDV among HBsAg chronic carriers from Maranhão state, a region located in the Northeast of Brazil. Among 133 patients, 5 had anti-HD, of whom 3 had HDV RNA. HDV genotypes were characterized by Bayesian phylogenetic analysis of nucleotide sequences from the HDAg coding region. HDV-3 was identified in one patient who lives in Maranhão, but was born in Amazonas state (Western Amazon basin). Phylogenetic analysis shows that this HDV-3 sequence grouped with other HDV-3 sequences isolated in this state, which suggests that the patient probably contracted HDV infection there. Surprisingly, the other two patients were infected with HDV-8, an African genotype. These patients were born and have always lived in Urbano Santos, a rural county of Maranhão state, moreover they had never been to Africa and denied any contact with people from that continent. This is the first description of the HDV-8 in non-native African populations. This genotype may have been introduced to Brazil through the slaves brought to the country from the West Africa regions during the 16-18th centuries. Our results indicate that the need of clinical and epidemiological studies to investigate the presence of this infection in other areas in Brazil. Copyright © 2011 Elsevier B.V. All rights reserved.

  1. Dengue Virus Type 4 Phylogenetics in Brazil 2011: Looking beyond the Veil

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    de Souza, Renato Pereira; Rocco, Iray M.; Maeda, Adriana Y.; Spenassatto, Carine; Bisordi, Ivani; Suzuki, Akemi; Silveira, Vivian R.; Silva, Sarai J. S.; Azevedo, Roberta M.; Tolentino, Fernanda M.; Assis, Jaqueline C.; Bassi, Margarida G.; Dambrós, Bibiana P.; Tumioto, Gabriela L.; Gregianini, Tatiana S.; Souza, Luiza Terezinha M.; Timenetsky, Maria do Carmo S. T.; Santos, Cecília L. S.

    2011-01-01

    Dengue Fever and Dengue Hemorrhagic Fever are diseases affecting approximately 100 million people/year and are a major concern in developing countries. In the present study, the phylogenetic relationship of six strains of the first autochthonous cases of DENV-4 infection occurred in Sao Paulo State, Parana State and Rio Grande do Sul State, Brazil, 2011 were studied. Nucleotide sequences of the envelope gene were determined and compared with sequences representative of the genotypes I, II, III and Sylvatic for DEN4 retrieved from GenBank. We employed a Bayesian phylogenetic approach to reconstruct the phylogenetic relationships of Brazilian DENV-4 and we estimated evolutionary rates and dates of divergence for DENV-4 found in Brazil in 2011. All samples sequenced in this study were located in Genotype II. The studied strains are monophyletic and our data suggest that they have been evolving separately for at least 4 to 6 years. Our data suggest that the virus might have been present in the region for some time, without being noticed by Health Surveillance Services due to a low level of circulation and a higher prevalence of DENV-1 and DENV- 2. PMID:22216365

  2. Exposure to hepatitis C virus in homeless men in Central Brazil: a cross-sectional study.

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    Ferreira, Priscilla Martins; Guimarães, Rafael Alves; Souza, Christiane Moreira; Guimarães, Lara Cristina da Cunha; Barros, Cleiciane Vieira de Lima; Caetano, Karlla Antonieta Amorim; Rezza, Giovanni; Spadoni, Lila; Brunini, Sandra Maria

    2017-01-18

    Homeless men are highly vulnerable to acquisition of the hepatitis C virus (HCV) compared to the general population. In Brazil, a country of continental dimensions, the extent of HCV infection in this population remains unknown. The objective of this study is to investigate the epidemiological profile of exposure to HCV in homeless men in Central Brazil. A Cross-sectional study was conducted in 481 men aged over 18 years attending therapeutic communities specialized in the recovery and reintegration of homeless people. Participants were tested for anti-HCV markers using rapid tests. Poisson regression analysis was used to verify the risk factors associated with exposure to HCV. The prevalence of HCV exposure was 2.5% (95.0% CI: 1.4 to 4.3%) and was associated with age, absence of family life, injection drug use, number of sexual partners, and history of sexually transmitted infections (STI). Participants reported multiple risk behaviors, such as alcohol (78.9%), cocaine (37.1%) and/or crack use (53.1%), and inconsistent condom use (82.6%). Injection drug use was reported by 8.7% of participants. The prevalence of HCV infection among homeless men was relatively high. Several risk behaviors were commonly reported, which shows the high vulnerability of this population. These findings emphasize the need for the development of specific strategies to reduce the risk of HCV among homeless men.

  3. Genetic characterization of St. Louis encephalitis virus isolated from human in São Paulo, Brazil

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    Cecília Luiza Simões dos Santos

    2006-02-01

    Full Text Available The molecular characterization of SPH253157, a new strain of St. Louis encephalitis virus (SLEV, isolated in 2004 from the first case of human infection recognized in the state of São Paulo, Brazil, is reported. The patient, presenting a febrile illness without neurological involvement, was hospitalized as a probable case of dengue fever. Genomic RNA was isolated from the supernatant of C6/36 cells infected with acute phase-serum specimen of the patient and the envelope gene was amplified by reverse-transcription-polymerase chain reaction. The complete nucleotide sequence of the envelope gene of this isolate was directly sequenced from the amplified products and compared with other Brazilian and American SLEV strains. Phylogenetic analyses were carried out under maximum likelihood criterion with outgroups both included and excluded. Outgroups comprised four flavivirus of the Japanese encephalitis group. Phylogeny also included Bayesian analysis. The results indicated that the new SLEV isolate belongs to lineage III, being closely related to an Argentinean strain recovered from Culex sp. in 1979. It is concluded that there are at least 3 lineages of SLEV in Brazil.

  4. Exposure to hepatitis C virus in homeless men in Central Brazil: a cross-sectional study

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    Priscilla Martins Ferreira

    2017-01-01

    Full Text Available Abstract Background Homeless men are highly vulnerable to acquisition of the hepatitis C virus (HCV compared to the general population. In Brazil, a country of continental dimensions, the extent of HCV infection in this population remains unknown. The objective of this study is to investigate the epidemiological profile of exposure to HCV in homeless men in Central Brazil. Methods A Cross-sectional study was conducted in 481 men aged over 18 years attending therapeutic communities specialized in the recovery and reintegration of homeless people. Participants were tested for anti-HCV markers using rapid tests. Poisson regression analysis was used to verify the risk factors associated with exposure to HCV. Results The prevalence of HCV exposure was 2.5% (95.0% CI: 1.4 to 4.3% and was associated with age, absence of family life, injection drug use, number of sexual partners, and history of sexually transmitted infections (STI. Participants reported multiple risk behaviors, such as alcohol (78.9%, cocaine (37.1% and/or crack use (53.1%, and inconsistent condom use (82.6%. Injection drug use was reported by 8.7% of participants. Conclusions The prevalence of HCV infection among homeless men was relatively high. Several risk behaviors were commonly reported, which shows the high vulnerability of this population. These findings emphasize the need for the development of specific strategies to reduce the risk of HCV among homeless men.

  5. V. Detection of the Virus in Infected Materials by Immunofluorescence

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    Ruckerbauer, G. M.; Gray, D. P.; Girard, A.; Bannister, G. L.; Boulanger, P.

    1967-01-01

    The fluorescent-antibody technique was employed for the detection of bluetongue virus in bovine foetal kidney cell cultures inoculated with tissues and blood from experimentally infected animals. In the first series, a total number of 79 inoculated suckling-mouse brains were examined, 36 as frozen sections alone and 43 as impression slides in conjunction with tissue culture inoculation of the same material. With the combined tissue culture immunofluorescent methods, 36 suspicious were detected by impression smears and 37 positives by the tissue culture out of 43 brains examined. Twenty-two were suspicious out of the 36 examined as frozen sections. Results obtained with the second series, using sheep tissues, showed that the combined tissue culture-fluorescent antibody method was satisfactory for demonstrating the virus in blood of infected animals 1 to 9 days postinfection, and in some organs after death. No false positive reactions were obtained. ImagesFig. 1. PMID:4227047

  6. Molecular detection of Bluetongue Virus (BTV and Bovine Leukemia Virus (BLV in uterine biopsies of dairy cows with or without reproductive problems

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    Juliana Marques Bicalho

    2016-10-01

    Full Text Available Reproductive performance of dairy cows has a direct impact on herd productivity. Infectious agents, such as Bluetongue Virus (BTV and Bovine Leukemia Virus (BLV, are associated with reproductive failure. However, it remains unknown if these viruses are present in the uterus and cause gestational loss. This study used molecular methods to assess if BTV and BLV can be detected in the uterus of serologically positive dairy cows with a record of abortions, stillbirths and repeat breeding (n=23 and without a record of reproductive problems (n =23. The cows came from three dairy herds of the state of Minas Gerais, Brazil. BTV was not detected in any of the uterine biopsies. Proviral DNA of BLV was detected in 54.5 % of the seropositive cows, but positivity for BLV in the uterus was not associated with the existence of reproductive problems. In conclusion, this study shows that BLV, but not BTV, is present in the uterus of seropositive cows, regardless of reproductive performance.

  7. Prolonged Detection of Zika Virus in Vaginal Secretions and Whole Blood

    Science.gov (United States)

    Gorchakov, Rodion; Carlson, Anna R.; Berry, Rebecca; Lai, Lilin; Natrajan, Muktha; Garcia, Melissa N.; Correa, Armando; Patel, Shital M.; Aagaard, Kjersti; Mulligan, Mark J.

    2017-01-01

    Infection with Zika virus is an emerging public health crisis. We observed prolonged detection of virus RNA in vaginal mucosal swab specimens and whole blood for a US traveler with acute Zika virus infection who had visited Honduras. These findings advance understanding of Zika virus infection and provide data for additional testing strategies. PMID:27748649

  8. Prolonged Detection of Zika Virus in Vaginal Secretions and Whole Blood.

    Science.gov (United States)

    Murray, Kristy O; Gorchakov, Rodion; Carlson, Anna R; Berry, Rebecca; Lai, Lilin; Natrajan, Muktha; Garcia, Melissa N; Correa, Armando; Patel, Shital M; Aagaard, Kjersti; Mulligan, Mark J

    2017-01-01

    Infection with Zika virus is an emerging public health crisis. We observed prolonged detection of virus RNA in vaginal mucosal swab specimens and whole blood for a US traveler with acute Zika virus infection who had visited Honduras. These findings advance understanding of Zika virus infection and provide data for additional testing strategies.

  9. Evaluation of techniques for the diagnosis of Strongyloides stercoralis in human immunodeficiency virus (HIV positive and HIV negative individuals in the city of Itajaí, Brazil

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    Jucelene Marchi Blatt

    Full Text Available Human immunodeficiency virus (HIV and intestinal parasites are common in Brazil. Previous studies have shown that infection with Strongyloides stercoralis is frequently associated with HIV infection. Strongyloidiasis is difficult to diagnosis and stool examination with conventional techniques fails to detect the helminth larvae. We made a prospective study, to test the efficacy of the agar plate technique to detect S. stercoralis in 211 HIV-positive patients and 213 HIV-negative patients in the city of Itajaí, Brazil, between September 2001 and June 2002. The feces samples of these patients were processed and analyzed according to the following methods: Lutz, formalin ethyl acetate, Baermann, Harada-Mori and agar plate culture. HIV-positive patients were more frequently infected by S. stercoralis (odds ratio= 5,.687. Among the methods used on fecal specimens, the larvae of S. stercoralis were most efficiently detected by the agar plate (69.7% method, followed by the Baermann and the formalin ethyl acetate methods (48.5% (P=0.01, Lutz (42.4% (P=0.01, and Harada-Mori culture (24% (P=0.001. Therefore agar plate culture is the most efficient method for the detection of S. stercoralis larvae and this technique should be the test of choice, especially in immunocompromised patients.

  10. Molecular detection of Aichi virus in raw sewage in Italy.

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    Di Martino, Barbara; Di Profio, Federica; Ceci, Chiara; Di Felice, Elisabetta; Marsilio, Fulvio

    2013-09-01

    Aichi virus (AiV) is suspected to play a role in viral gastroenteritis in humans. In this study, we assessed the presence of AiV in untreated influent sewage samples collected at four wastewater treatment plants in central Italy. AiV was detected in 6 (12.5 %) of the 48 specimens and in all plants. All of the Italian strains showed the highest nucleotide and amino acid sequence identity to genotype B AiV detected recently in Asia, especially in China.

  11. Recurrent Potent Human Neutralizing Antibodies to Zika Virus in Brazil and Mexico.

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    Robbiani, Davide F; Bozzacco, Leonia; Keeffe, Jennifer R; Khouri, Ricardo; Olsen, Priscilla C; Gazumyan, Anna; Schaefer-Babajew, Dennis; Avila-Rios, Santiago; Nogueira, Lilian; Patel, Roshni; Azzopardi, Stephanie A; Uhl, Lion F K; Saeed, Mohsan; Sevilla-Reyes, Edgar E; Agudelo, Marianna; Yao, Kai-Hui; Golijanin, Jovana; Gristick, Harry B; Lee, Yu E; Hurley, Arlene; Caskey, Marina; Pai, Joy; Oliveira, Thiago; Wunder, Elsio A; Sacramento, Gielson; Nery, Nivison; Orge, Cibele; Costa, Federico; Reis, Mitermayer G; Thomas, Neena M; Eisenreich, Thomas; Weinberger, Daniel M; de Almeida, Antonio R P; West, Anthony P; Rice, Charles M; Bjorkman, Pamela J; Reyes-Teran, Gustavo; Ko, Albert I; MacDonald, Margaret R; Nussenzweig, Michel C

    2017-05-04

    Antibodies to Zika virus (ZIKV) can be protective. To examine the antibody response in individuals who develop high titers of anti-ZIKV antibodies, we screened cohorts in Brazil and Mexico for ZIKV envelope domain III (ZEDIII) binding and neutralization. We find that serologic reactivity to dengue 1 virus (DENV1) EDIII before ZIKV exposure is associated with increased ZIKV neutralizing titers after exposure. Antibody cloning shows that donors with high ZIKV neutralizing antibody titers have expanded clones of memory B cells that express the same immunoglobulin VH3-23/VK1-5 genes. These recurring antibodies cross-react with DENV1, but not other flaviviruses, neutralize both DENV1 and ZIKV, and protect mice against ZIKV challenge. Structural analyses reveal the mechanism of recognition of the ZEDIII lateral ridge by VH3-23/VK1-5 antibodies. Serologic testing shows that antibodies to this region correlate with serum neutralizing activity to ZIKV. Thus, high neutralizing responses to ZIKV are associated with pre-existing reactivity to DENV1 in humans. Copyright © 2017 Elsevier Inc. All rights reserved.

  12. Prevalence and clinical features of respiratory syncytial virus in children hospitalized for community-acquired pneumonia in northern Brazil

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    Lamarão Letícia

    2012-05-01

    Full Text Available Abstract Background Childhood pneumonia and bronchiolitis is a leading cause of illness and death in young children worldwide with Respiratory Syncytial Virus (RSV as the main viral cause. RSV has been associated with annual respiratory disease outbreaks and bacterial co-infection has also been reported. This study is the first RSV epidemiological study in young children hospitalized with community-acquired pneumonia (CAP in Belém city, Pará (Northern Brazil. Methods With the objective of determining the prevalence of RSV infection and evaluating the patients’ clinical and epidemiological features, we conducted a prospective study across eight hospitals from November 2006 to October 2007. In this study, 1,050 nasopharyngeal aspirate samples were obtained from hospitalized children up to the age of three years with CAP, and tested for RSV antigen by direct immunofluorescence assay and by Reverse Transcription Polymerase Chain Reaction (RT-PCR for RSV Group identification. Results RSV infection was detected in 243 (23.1% children. The mean age of the RSV-positive group was lower than the RSV-negative group (12.1 months vs 15.5 months, pppppp Conclusion The present study highlights the relevance of RSV infection in hospitalized cases of CAP in our region; our findings warrant the conduct of further investigations which can help design strategies for controlling the disease.

  13. Molecular detection of enteric viruses from diarrheic calves in Egypt.

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    Mohamed, Fakry F; Mansour, Shimaa M G; El-Araby, Iman E; Mor, Sunil K; Goyal, Sagar M

    2017-01-01

    Neonatal calf diarrhea (NCD) is a major cause of morbidity, mortality and economic losses in the beef and dairy industries. This study was conducted to investigate the existence of enteric viruses in two Egyptian farms with a history of recurrent diarrhea. Fecal samples were collected from 25 diarrheic calves. RNA was extracted and tested by reverse transcription polymerase chain reaction (RT-PCR) for the presence of rotavirus, norovirus, astrovirus, torovirus, coronavirus and bovine viral diarrhea virus. Overall, 76 % (19/25) of samples tested positive for one or more viruses. Rota-, noro- and astroviruses were detected in 48 %, 24 % and 32 % of tested samples, respectively. About 37 % (7/19) of positive samples had two different viruses. One-month-old calves were the group most vulnerable to infections. Based on phylogenetic analysis, bovine rotaviruses were of genotypes G6 and G10, bovine noroviruses were in GIII.2, and bovine astroviruses were in the BAstV lineage 1. Astrovirus sequences showed a high level nucleotide sequence similarity with the Brazilian BAstV sequences available in GenBank. We believe this is the first report of bovine norovirus and bovine astrovirus circulating among calves in Egypt. Further epidemiological studies are recommended to investigate their presence on a wider scale, to predict their association with NCD, and to design appropriate diagnostic and control methods.

  14. Notification of the first isolation of Cacipacore virus in a human in the State of Rondônia, Brazil

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    Weber Cheli Batista

    2011-08-01

    Full Text Available Flavivirus is a genus of arthropod-transmitted viruses of the family Flaviviridae, and in Brazil, up to eleven different Flavivirus have been isolated. We collected blood from farmers in the municipality of Theobroma, which is located 320km from the City of Porto Velho, the former capital of the Brazilian State of Rondônia. For viral isolation, we used newborn mouse brain, followed by RT-PCR with specific universal Flavivirus primers. We obtained fragments 958bp and 800bp in length. Based on BLAST, these sequences were 91% similar to a sequence of Cacipacore virus.

  15. Rapid and specific detection of Asian- and African-lineage Zika viruses

    Science.gov (United States)

    Chotiwan, Nunya; Brewster, Connie D.; Magalhaes, Tereza; Weger-Lucarelli, James; Duggal, Nisha K.; Rückert, Claudia; Nguyen, Chilinh; Garcia Luna, Selene M.; Fauver, Joseph R.; Andre, Barb; Gray, Meg; Black, William C.; Kading, Rebekah C.; Ebel, Gregory D.; Kuan, Guillermina; Balmaseda, Angel; Jaenisch, Thomas; Marques, Ernesto T. A.; Brault, Aaron C.; Harris, Eva; Foy, Brian D.; Quackenbush, Sandra L.; Perera, Rushika; Rovnak, Joel

    2017-01-01

    Understanding the dynamics of Zika virus transmission and formulating rational strategies for its control require precise diagnostic tools that are also appropriate for resource-poor environments. We have developed a rapid and sensitive loop-mediated isothermal amplification (LAMP) assay that distinguishes Zika viruses of Asian and African lineages. The assay does not detect chikungunya virus or flaviviruses such as dengue, yellow fever, or West Nile viruses. The assay conditions allowed direct detection of Zika virus RNA in cultured infected cells; in mosquitoes; in virus-spiked samples of human blood, plasma, saliva, urine, and semen; and in infected patient serum, plasma, and semen samples without the need for RNA isolation or reverse transcription. The assay offers rapid, specific, sensitive, and inexpensive detection of the Asian-lineage Zika virus strain that is currently circulating in the Western hemisphere, and can also detect the African-lineage Zika virus strain using separate, specific primers. PMID:28469032

  16. Influenza and respiratory syncytial virus are the major respiratory viruses detected from prospective testing of pediatric and adult coronial autopsies.

    Science.gov (United States)

    Speers, David J; Moss, Daniel M; Minney-Smith, Cara; Levy, Avram; Smith, David W

    2013-11-01

    To ascertain the full mortality of influenza and other respiratory viruses, the testing of community autopsy specimens is essential. Respiratory virus PCR and culture were performed on 2418 fresh unfrozen respiratory samples collected from 1611 coronial cases where the death was either unknown or infection was suspected, from July 2007 to June 2011, to detect the common respiratory viruses in children and adults, using standardized microbiological testing. The respiratory virus positive rate was 8·3% (134 cases) with a peak of 28% (42 of 151 cases) in children under 10 years of age. Influenza virus was the commonest respiratory virus (50 cases, 3%), followed by respiratory syncytial virus (RSV) (30 cases, 2%). All tested respiratory viruses were found in children, most commonly adenovirus, enterovirus and RSV, and influenza A and RSV predominated in those over 60 years, but coinfection was uncommon. Almost all influenza cases occurred when influenza was widely circulating in the community but few were diagnosed pre-mortem. Influenza and RSV detection was associated with bronchitis or bronchiolitis in 7 (9%) of the 80 cases and caused pneumonia in 14 (0·8%) deaths overall. Our prospective review of respiratory viruses using standardized testing found a single lower respiratory tract autopsy specimen for respiratory virus PCR would detect most community infections at the time of death. © 2013 John Wiley & Sons Ltd.

  17. Detection of dengue group viruses by fluorescence in situ hybridization

    Science.gov (United States)

    2012-01-01

    Background Dengue fever (DF) and dengue hemorrhagic fever (DHF) represent a global challenge in public health. It is estimated that 50 to 100 million infections occur each year causing approximately 20,000 deaths that are usually linked to severe cases like DHF and dengue shock syndrome. The causative agent of DF is dengue virus (genus Flavivirus) that comprises four distinct serotypes (DENV-1 to DENV-4). Fluorescence in situ hybridization (FISH) has been used successfully to detect pathogenic agents, but has not been implemented in detecting DENV. To improve our understanding of DENV infection and dissemination in host tissues, we designed specific probes to detect DENV in FISH assays. Methods Oligonucleotide probes were designed to hybridize with RNA from the broadest range of DENV isolates belonging to the four serotypes, but not to the closest Flavivirus genomes. Three probes that fit the criteria defined for FISH experiments were selected, targeting both coding and non-coding regions of the DENV genome. These probes were tested in FISH assays against the dengue vector Aedes albopictus (Diptera: Culicidae). The FISH experiments were led in vitro using the C6/36 cell line, and in vivo against dissected salivary glands, with epifluorescence and confocal microscopy. Results The three 60-nt oligonucleotides probes DENV-Probe A, B and C cover a broad range of DENV isolates from the four serotypes. When the three probes were used together, specific fluorescent signals were observed in C6/36 infected with each DENV serotypes. No signal was detected in either cells infected with close Flavivirus members West Nile virus or yellow fever virus. The same protocol was used on salivary glands of Ae. albopictus fed with a DENV-2 infectious blood-meal which showed positive signals in the lateral lobes of infected samples, with no significant signal in uninfected mosquitoes. Conclusion Based on the FISH technique, we propose a way to design and use oligonucleotide probes to

  18. Application of artificial intelligence for detecting derived viruses

    CSIR Research Space (South Africa)

    Asiru, OF

    2017-06-01

    Full Text Available of artificial intelligence in detecting computing viruses (Gang & Zhongquan, 2014). However, more research still needs to be conducted in order to fully explore options from the field of artificial intelligence. Furthermore, few research efforts focus... stream_source_info Asiru_19173_2017.pdf.txt stream_content_type text/plain stream_size 35251 Content-Encoding UTF-8 stream_name Asiru_19173_2017.pdf.txt Content-Type text/plain; charset=UTF-8 Application of Artificial...

  19. Acute hepatitis C virus infection assessment among chronic hemodialysis patients in the Southwest Parana State, Brazil

    Directory of Open Access Journals (Sweden)

    Ono-Nita Suzane K

    2007-04-01

    Full Text Available Abstract Background Chronic hemodialysis patients are at higher risk for acquiring hepatitis C virus (HCV. The prevalence varies among different countries and hemodialysis centers. Although guidelines for a comprehensive infection control program exist, the nosocomial transmission still accounts for the new cases of infection. The aim of this study was analyze the follow up of newly acquired acute hepatitis C cases, during the period from January 2002 to May 2005, in the Hemodialysis Center, located in the Southwest region of Parana State, Brazil and to analyze the effectiveness of the measures to restrain the appearance of new cases of acute hepatitis C. Methods Patients were analyzed monthly with anti-HCV tests and ALT measurements. Patients with ALT elevations were monitored for possible acute hepatitis C. Results During this period, 32 new cases were identified with acute hepatitis C virus infection. Blood screening showed variable ALT levels preceding the anti-HCV seroconversion. HCV RNA viremia by PCR analysis was intermittently and even negative in some cases. Ten out of 32 patients received 1 mcg/kg dose of pegylated interferon alfa-2b treatment for 24 weeks. All dialysis personnel were re-trained to strictly follow the regulations and recommendations regarding infection control, proper methods to clean and disinfect equipment were reviewed and HCV-positive patients were isolated. Conclusion Laboratory tests results showed variable ALT preceding anti-HCV seroconversion and intermittent viremia. The applied recommendations contributed importantly to restrain the appearance of new cases of acute hepatitis C in this center and the last case was diagnosed in May 2004.

  20. Enteric viruses in surface waters from Argentina: molecular and viable virus detection.

    Science.gov (United States)

    Masachessi, G; Pisano, M B; Prez, V E; Martínez, L C; Michelena, J F; Martínez-Wassaf, M; Giordano, M O; Isa, M B; Pavan, J V; Welter, A; Nates, S V; Ré, V

    2017-12-21

    The water resources contaminated with wastewater are an important source of dissemination of enteric viruses with impact on the health of the populations. The aim of the study was to assess the viral contamination of freshwater from a dam in Argentina, including infectious enterovirus detection, viral RNA amplification and genetic characterization of five enteric viruses associated to diarrhea and hepatitis. Enterovirus infectivity (iEV) was evaluated by cell culture and direct immunofluorescence. Detection of viral genome of rotavirus (RV), human astrovirus (HAstV), norovirus (NoV), hepatitis A (HAV) and hepatitis E (HEV) viruses, was performed by RT-PCR. A total of 48 water samples from 4 monitoring points on the body of the Dam from January to December 2012 and 66 water samples from 3 tourist beaches on the edge of the dam from October 2013 to October 2015 were monthly collected. During the first period of time, the overall viral frequency detection was 52.1% for RVA, 50% for HAstV, 60.4% for NoV, 22.9% for HAV, 2.1% for HEV and 64.6% for iEV. The overall frequency detection for the second sampling was 18.2 % for RV and HAstV, for NoV 31.8%, for HEV 0.76% and for iEV 66.7%. There was no detection of HAV during this period. The genotypes and genogroups detected through the study, correlated to the most common genomic variants associated with human gastrointestinal and hepatitis illness. The results obtained alert the health systems and environmental sanitation in order to make decisions for viral control and prevention in our environment.Importance The study shows the impact of anthropic contamination of one of the most important tourist water resources in our country. This course of recreational water would be a favorable scenario for the infection, as well as a reservoir for the enteric viruses, creating a risk for the population exposed to these waters. The results obtained could alert the health systems and environmental sanitation in order to make decisions

  1. Two novel circo-like viruses detected in human feces: complete genome sequencing and electron microscopy analysis.

    Science.gov (United States)

    Castrignano, Silvana Beres; Nagasse-Sugahara, Teresa Keico; Kisielius, Jonas José; Ueda-Ito, Marli; Brandão, Paulo Eduardo; Curti, Suely Pires

    2013-12-26

    The application of viral metagenomic techniques and a series of PCRs in a human fecal sample enabled the detection of two novel circular unisense DNA viral genomes with 92% nucleotide similarity. The viruses were tentatively named circo-like virus-Brazil (CLV-BR) strains hs1 and hs2 and have genome lengths of 2526 and 2533 nucleotides, respectively. Four major open reading frames (ORFs) were identified in each of the genomes, and differences between the two genomes were primarily observed in ORF 2. Only ORF 3 showed significant amino acid similarities to a putative rolling circle replication initiator protein (Rep), although with low identity (36%). Our phylogenetic analysis, based on the Rep protein, demonstrated that the CLV-BRs do not cluster with members of the Circoviridae, Nanoviridae or Geminiviridae families and are more closely related to circo-like genomes previously identified in reclaimed water and feces of a wild rodent and of a bat. The CLV-BRs are members of a putative new family of circular Rep-encoding ssDNA viruses. Electron microscopy revealed icosahedral (~23 nm) structures, likely reflecting the novel viruses, and rod-shaped viral particles (~65-460 × 21 × 10 nm in length, diameter, and axial canal, respectively). Circo-like viruses have been detected in stool samples from humans and other mammals (bats, rodents, chimpanzees and bovines), cerebrospinal fluid and sera from humans, as well as samples from many other sources, e.g., insects, meat and the environment. Further studies are needed to classify all novel circular DNA viruses and elucidate their hosts, pathogenicity and evolutionary history. Copyright © 2013 Elsevier B.V. All rights reserved.

  2. Insect-specific flaviviruses, a worldwide widespread group of viruses only detected in insects.

    Science.gov (United States)

    Calzolari, Mattia; Zé-Zé, Líbia; Vázquez, Ana; Sánchez Seco, Mari Paz; Amaro, Fátima; Dottori, Michele

    2016-06-01

    Several flaviviruses are important pathogens for humans and animals (Dengue viruses, Japanese encephalitis virus, Yellow-fever virus, Tick-borne encephalitis virus, West Nile virus). In recent years, numerous novel and related flaviviruses without known pathogenic capacity have been isolated worldwide in the natural mosquito population. However, phylogenetic studies have shown that genomic sequences of these viruses diverge from other flaviviruses. Moreover, these viruses seem to be exclusive of insects (they do not seem to grow on vertebrate cell lines), and were already defined as mosquito-only flaviviruses or insect-specific flaviviruses. At least eleven of these viruses were isolated worldwide, and sequences ascribable to other eleven putative viruses were detected in several mosquito species. A large part of the cycle of these viruses is not well known, and their persistence in the environment is poorly understood. These viruses are detected in a wide variety of distinct mosquito species and also in sandflies and chironomids worldwide; a single virus, or the genetic material ascribable to a virus, was detected in several mosquito species in different countries, often in different continents. Furthermore, some of these viruses are carried by invasive mosquitoes, and do not seem to have a depressive action on their fitness. The global distribution and the continuous detection of new viruses in this group point out the likely underestimation of their number, and raise interesting issues about their possible interactions with the pathogenic flaviviruses, and their influence on the bionomics of arthropod hosts. Some enigmatic features, as their integration in the mosquito genome, the recognition of their genetic material in DNA forms in field-collected mosquitoes, or the detection of the same virus in both mosquitoes and sandflies, indicate that the cycle of these viruses has unknown characteristics that could be of use to reach a deeper understanding of the cycle

  3. The microbial detection array for detection of emerging viruses in clinical samples--a useful panmicrobial diagnostic tool

    DEFF Research Database (Denmark)

    Rosenstierne, Maiken W; McLoughlin, Kevin S; Olesen, Majken Lindholm

    2014-01-01

    Emerging viruses are usually endemic to tropical and sub-tropical regions of the world, but increased global travel, climate change and changes in lifestyle are believed to contribute to the spread of these viruses into new regions. Many of these viruses cause similar disease symptoms as other...... emerging viruses or common infections, making these unexpected pathogens difficult to diagnose. Broad-spectrum pathogen detection microarrays containing probes for all sequenced viruses and bacteria can provide rapid identification of viruses, guiding decisions about treatment and appropriate case...... management. We report a modified Whole Transcriptome Amplification (WTA) method that increases unbiased amplification, particular of RNA viruses. Using this modified WTA method, we tested the specificity and sensitivity of the Lawrence Livermore Microbial Detection Array (LLMDA) against a wide range...

  4. The dynamics of dengue virus serotype 3 introduction and dispersion in the state of Bahia, Brazil

    Directory of Open Access Journals (Sweden)

    Paulo Roberto Santana de Melo

    2007-12-01

    Full Text Available By 2002, dengue virus serotype 1 (DENV-1 and DENV-2 had circulated for more than a decade in Brazil. In 2002, the introduction of DENV-3 in the state of Bahia produced a massive epidemic and the first cases of dengue hemorrhagic fever. Based on the standardized frequency, timing and location of viral isolations by the state's Central Laboratory, DENV-3 probably entered Bahia through its capital, Salvador, and then rapidly disseminated to other cities, following the main roads. A linear regression model that included traffic flow, distance from the capital and DENV-1 circulation (r² = 0.24, p = 0.001 supported this hypothesis. This pattern was not seen for serotypes already in circulation and was not seen for DENV-3 in the following year. Human population density was another important factor in the intensity of viral circulation. Neither DENV-1 nor DENV-2 fit this model for 2001 or 2003. Since the vector has limited flight range and vector densities fail to correlate with intensity of viral circulation, this distribution represents the movement of infected people and to some extent mosquitoes. This pattern may mimic person-to-person spread of a new infection.

  5. Detection of enteroviruses in cases of neurological disorders in the State of Pará, Brazil

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    GOMES Maria de Lourdes Contente

    2001-01-01

    Full Text Available Eighty-one cerebrospinal fluid (CSF samples mainly from cases of aseptic meningitis and motor deficiency syndrome were sent to the Virology Section of Evandro Chagas Institute, Belém Pará, in the period of January 1995 to January 1996 in order to isolate viruses. All samples were inoculated onto HEp-2 cell culture and newborn mice, with negative results. The probability of isolating viruses by these methods is reduced because of the low concentration of viral particles in these specimens. In order to obtain more information about the etiology of these cases, a group of 23 samples were selected to be tested by a more sensitive technique than the virus isolation - the reverse transcription polymerase chain reaction (RT-PCR. Specific primers directed to conserved regions in the enterovirus genome were used, considering that this group of viruses is frequently associated with these neurological disorder. The age of the patients ranged from 1 to 55 years and nearly all of them lived in Belém, State of Pará, North of Brazil. Of 15 samples analyzed by RT PCR nine (60% were positive; of these, 6 (66.6% had motor deficiency and 3 (33.3% developed aseptic meningitis. These results show that it is important to investigate enterovirus as cause of these syndromes.

  6. Detection of three Allexivirus species infecting garlic in Brazil Detecção de três espécies de Allexivirus que infectam o alho no Brasil

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    Péricles de Albuquerque Melo Filho

    2004-08-01

    Full Text Available Garlic viruses often occur in mixed infections under field conditions. In this study, garlic samples collected in three geographical areas of Brazil were tested by Dot-ELISA for the detection of allexiviruses using monoclonal specific antibodies to detect Garlic virus A (GarV-A, Garlic virus B (GarV-B, Garlic virus C (GarV-C and a polyclonal antiserum able to detect the three virus species mentioned plus Garlic virus D (GarV-D. The detected viruses were biologically isolated by successive passages through Chenopodium quinoa. Reverse Transcriptase Polimerase Chain Reaction (RT-PCR was performed using primers designed from specific regions of the coat protein genes of Japanese allexiviruses available in the Genetic Bank of National Center of Biotechnology Information (NCBI. By these procedures, individual garlic virus genomes were isolated and sequenced. The nucleotide and amino acid sequence analysis and the one with serological data revealed the presence of three distinct allexiviruses GarV-C, GarV-D and a recently described allexivirus, named Garlic mite-borne filamentous virus (GarMbFV, in Brazil.Infecções virais em alho são normalmente causadas por um complexo viral. Neste estudo, um complexo viral de alho, coletado em campo, em três regiões geográficas, foi testado com anti-soros monoclonais específicos para Garlic virus A (GarV-A, Garlic virus B (GarV-B, Garlic virus C (GarV-C e um anti-soro policlonal capaz de detectar os três vírus mencionados e Garlic virus D (GarV-D. Procedeu-se à amplificação por transcriptase reversa-reação em cadeia da polimerase (RT-PCR usando oligonucleotídeos sintetizados a partir de regiões específicas de genes de proteínas capsidiais de allexivirus japoneses e disponíveis no GeneBank (National Center of Biotechnology Information - NCBI. Por esse procedimento, vírus individuais foram isolados e seqüenciados. Os vírus detectados foram biologicamente isolados por meio de sucessivas inocula

  7. Antibody Derived Peptides for Detection of Ebola Virus Glycoprotein.

    Directory of Open Access Journals (Sweden)

    Luis Mario Rodríguez-Martínez

    Full Text Available Current Ebola virus (EBOV detection methods are costly and impractical for epidemic scenarios. Different immune-based assays have been reported for the detection and quantification of Ebola virus (EBOV proteins. In particular, several monoclonal antibodies (mAbs have been described that bind the capsid glycoprotein (GP of EBOV GP. However, the currently available platforms for the design and production of full-length mAbs are cumbersome and costly. The use of antibody fragments, rather than full-length antibodies, might represent a cost-effective alternative for the development of diagnostic and possibly even therapeutic alternatives for EBOV.We report the design and expression of three recombinant anti-GP mAb fragments in Escherichia coli cultures. These fragments contained the heavy and light variable portions of the three well-studied anti-GP full-length mAbs 13C6, 13F6, and KZ52, and are consequently named scFv-13C6, scFv-13F6, and Fab-KZ52, respectively. All three fragments exhibited specific anti-GP binding activity in ELISA experiments comparable to that of full-length anti-GP antibodies (i.e., the same order of magnitude and they are easily and economically produced in bacterial cultures.Antibody fragments might represent a useful, effective, and low cost alternative to full-length antibodies in Ebola related capture and diagnostics applications.

  8. [Characterization of hepatitis C virus in chronic hepatitis patients: genotypes in the State of Amazonas, Brazil].

    Science.gov (United States)

    Araújo, Ana Ruth; Almeida, Carlos Mauríco de; Fraporti, Liziara; Garcia, Nadja; Lima, Tatiane Amábili de; Maia, Laura Patrícia Viana; Torres, Kátia Luz; Tarragô, Andréa Monteiro; Victória, Flamir; Victória, Marilu; Tateno, Adriana; Levi, José Eduardo; Talhari, Sinésio; Malheiro, Adriana

    2011-10-01

    In the State of Amazonas, data regarding the prevalence of different genotypes of hepatitis C virus remains scarce. The genotype of 69 HCV positive patients was determined. An in-house standardized nested-PCR was used to detect HCV RNA. Genotype assignment was based on type-specific motifs on the sequenced amplicons delimited by primers HC11/HC18 from the 5' untranslated region. Of the 69 patients studied, 65.2% were male and 34.8% were female. Genotype 1 showed the greatest prevalence, followed by 3 and 2. These data suggesting that Manaus is the point of arrival of HCV in the State of Amazonas.

  9. Detection of respiratory syncytial virus and rhinovirus in healthy infants.

    Science.gov (United States)

    Hasegawa, Kohei; Linnemann, Rachel W; Avadhanula, Vasanthi; Mansbach, Jonathan M; Piedra, Pedro A; Gern, James E; Camargo, Carlos A

    2015-11-25

    Despite the research importance of rhinovirus detection in asymptomatic healthy infants, the literature remains sparse. To investigate the prevalence of respiratory syncytial virus (RSV) and rhinovirus (and its species). We conducted a cross-sectional study of 110 healthy, non-hospitalized infants without acute illness at an academic medical center from November 2013 through May 2014. We tested nasal swab specimens by using polymerase chain reaction and genetic sequencing. Overall, the median age was 3.8 months (IQR 2.0-5.1 months), 56 % were male, and 90% were born >37 weeks. RSV was detected in nasal swabs from infants (1.8%). By contrast, rhinovirus was detected in nasal swabs from 16 infants (14.5%). Molecular typing assay revealed rhinovirus species: six rhinovirus-A (5.5%), one rhinovirus-B (0.9%), eight rhinovirus-C (7.3%), and one untypeable (0.9%). In this cross-sectional study of healthy, community-based infants, RSV was rare (rhinovirus was detected in 14.5% with a predominance of rhinovirus-A and -C. These finding are important for understanding the clinical significance of rhinovirus detection among infants hospitalized for bronchiolitis.

  10. Detection and serological identification of adeno-associated virus in avian adenovirus stocks.

    Science.gov (United States)

    El Mishad, A M; McCormick, K J; Yates, V J; Trentin, J J

    1975-02-01

    Eleven avian adenovirus strains were tested for the presence of avian adeno-associated viruses (AAAV). Six strains contained AAAV. Electron microscopy using rabbit anti-AAAV serum was useful in detecting the satellite virus. The AAAV previously isolated from guail bronchitis virus was related to each of the six new isolates by immunoagglutination, complement fixation, immunodiffusion, and neutralization tests.

  11. Molecular detection and typing of influenza viruses. Are we ready for an influenza pandemic?

    NARCIS (Netherlands)

    MacKay, W.G.; Loon, A.M. van; Niedrig, M.; Meijer, A.; Lina, B.; Niesters, H.G.M.

    2008-01-01

    BACKGROUND: We cannot predict when an influenza pandemic will occur or which variant of the virus will cause it. Little information is currently available on the ability of laboratories to detect and subtype influenza viruses including the avian influenza viruses. OBJECTIVES: To assess the ability

  12. 9 CFR 113.55 - Detection of extraneous agents in Master Seed Virus.

    Science.gov (United States)

    2010-01-01

    ... Master Seed Virus. 113.55 Section 113.55 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION... STANDARD REQUIREMENTS Ingredient Requirements § 113.55 Detection of extraneous agents in Master Seed Virus... Seed Virus (MSV) shall be tested as prescribed in this section. A MSV found unsatisfactory by any...

  13. International Standardisation of a Method for Detection of Human Pathogenic Viruses in Molluscan Shellfish

    DEFF Research Database (Denmark)

    Lees, David; Schultz, Anna Charlotte

    2010-01-01

    The viruses primarily associated with shellfish-borne illness are norovirus, causing gastroenteritis and hepatitis A virus (HAV). Recent years have seen a proliferation of publications on methods for detection of these viruses in shellfish using polymerase chain reaction (PCR). However, currently...

  14. Charge detection mass spectrometry: Instrumentation & applications to viruses

    Science.gov (United States)

    Pierson, Elizabeth E.

    For over three decades, electrospray ionization (ESI) has been used to ionize non-covalent complexes and subsequently transfer the intact ion into the gas phase for mass spectrometry (MS) analysis. ESI generates a distribution of multiple charged ions, resulting in an m/z spectrum comprised of a series of peaks, known as a charge state envelope. To obtain mass information, the number of charges for each peak must be deduced. For smaller biological analytes like peptides, the charge states are sufficiently resolved and this process is straightforward. For macromolecular complexes exceeding ~100 kDa, this process is complicated by the broadening and shifting of charge states due to incomplete desolvation, salt adduction, and inherent mass heterogeneity. As the analyte mass approaches the MDa regime, the m/z spectrum is often comprised of a broad distribution of unresolved charge states. In such cases, mass determination is precluded. Charge detection mass spectrometry (CDMS) is an emerging MS technique for determining the masses of heterogeneous, macromolecular complexes. In CDMS, the m/z and z of single ions are measured concurrently so that mass is easily calculated. With this approach, deconvolution of an m/z spectrum is unnecessary. This measurement is carried out by passing macroions through a conductive cylinder. The induced image charge on the cylindrical detector provides information about m/z and z: the m/z is related to its time-of-flight through the detector, and the z is related to the intensity of the image charge. We have applied CDMS to study the self-assembly of virus capsids. Late-stage intermediates in the assembly of hepatitis B virus, a devastating human pathogen, have been identified. This is the first time that such intermediates have been detected and represent a significant advancement towards understanding virus capsid assembly. CDMS has also been used to identify oversized, non-icosahedral polymorphs in the assembly of woodchuck hepatitis

  15. Lateral Flow Immunoassays for Ebola Virus Disease Detection in Liberia.

    Science.gov (United States)

    Phan, Jill C; Pettitt, James; George, Josiah S; Fakoli, Lawrence S; Taweh, Fahn M; Bateman, Stacey L; Bennett, Richard S; Norris, Sarah L; Spinnler, David A; Pimentel, Guillermo; Sahr, Phillip K; Bolay, Fatorma K; Schoepp, Randal J

    2016-10-15

     Lateral flow immunoassays (LFIs) are point-of-care diagnostic assays that are designed for single use outside a formal laboratory, with in-home pregnancy tests the best-known example of these tests. Although the LFI has some limitations over more-complex immunoassay procedures, such as reduced sensitivity and the potential for false-positive results when using complex sample matrices, the assay has the benefits of a rapid time to result and ease of use. These benefits make it an attractive option for obtaining rapid results in an austere environment. In an outbreak of any magnitude, a field-based rapid diagnostic assay would allow proper patient transport and for safe burials to be conducted without the delay caused by transport of samples between remote villages and testing facilities. Use of such point-of-care instruments in the ongoing Ebola virus disease (EVD) outbreak in West Africa would have distinct advantages in control and prevention of local outbreaks, but proper understanding of the technology and interpretation of results are important.  In this study, a LFI, originally developed by the Naval Medical Research Center for Ebola virus environmental testing, was evaluated for its ability to detect the virus in clinical samples in Liberia. Clinical blood and plasma samples and post mortem oral swabs submitted to the Liberian Institute for Biomedical Research, the National Public Health Reference Laboratory for EVD testing, were tested and compared to results of real-time reverse transcription-polymerase chain reaction (rRT-PCR), using assays targeting Ebola virus glycoprotein and nucleoprotein.  The LFI findings correlated well with those of the real-time RT-PCR assays used as benchmarks.  Rapid antigen-detection tests such as LFIs are attractive alternatives to traditional immunoassays but have reduced sensitivity and specificity, resulting in increases in false-positive and false-negative results. An understanding of the strengths, weaknesses

  16. Optimizing a custom tiling microarray for low input detection and identification of unamplified virus targets.

    Science.gov (United States)

    Yu, Christine; Wales, Samantha Q; Mammel, Mark K; Hida, Kaoru; Kulka, Michael

    2016-08-01

    Viruses are major pathogens causing foodborne illnesses and are often present at low levels in foods, thus requiring sensitive techniques for their detection in contaminated foods. The lack of efficient culture methods for many foodborne viruses and the potential for multi-species viral contamination have driven investigation toward non-amplification based methods for virus detection and identification. A custom DNA microarray (FDA_EVIR) was assessed for its sensitivity in the detection and identification of low-input virus targets, human hepatitis A virus, norovirus, and coxsackievirus, individually and in combination. Modifications to sample processing were made to accommodate low input levels of unamplified virus targets, which included addition of carrier cDNA, RNase treatment, and optimization of DNase I-mediated target fragmentation. Amplification-free detection and identification of foodborne viruses were achieved in the range of 250-500 copies of virus RNA. Alternative data analysis methods were employed to distinguish the genotypes of the viruses particularly at lower levels of target input and the single probe-based analysis approach made it possible to identify a minority species in a multi-virus complex. The oligonucleotide array is shown to be a promising platform to detect foodborne viruses at low levels close to what are anticipated in food or environmental samples. Published by Elsevier B.V.

  17. Dengue virus type 3 adaptive changes during epidemics in São Jose de Rio Preto, Brazil, 2006-2007.

    Directory of Open Access Journals (Sweden)

    Christian Julian Villabona-Arenas

    Full Text Available Global dengue virus spread in tropical and sub-tropical regions has become a major international public health concern. It is evident that DENV genetic diversity plays a significant role in the immunopathology of the disease and that the identification of polymorphisms associated with adaptive responses is important for vaccine development. The investigation of naturally occurring genomic variants may play an important role in the comprehension of different adaptive strategies used by these mutants to evade the human immune system. In order to elucidate this role we sequenced the complete polyprotein-coding region of thirty-three DENV-3 isolates to characterize variants circulating under high endemicity in the city of São José de Rio Preto, Brazil, during the onset of the 2006-07 epidemic. By inferring the evolutionary history on a local-scale and estimating rates of synonymous (dS and nonsynonimous (dN substitutions, we have documented at least two different introductions of DENV-3 into the city and detected 10 polymorphic codon sites under significant positive selection (dN/dS > 1 and 8 under significant purifying selection (dN/dS < 1. We found several polymorphic amino acid coding sites in the envelope (15, NS1 (17, NS2A (11, and NS5 (24 genes, which suggests that these genes may be experiencing relatively recent adaptive changes. Furthermore, some polymorphisms correlated with changes in the immunogenicity of several epitopes. Our study highlights the existence of significant and informative DENV variability at the spatio-temporal scale of an urban outbreak.

  18. Detection and tentative grouping of Strawberry crinkle virus isolates

    NARCIS (Netherlands)

    Klerks, M.M.; Lindner, J.L.; Vasková, D.; Spak, J.; Thompson, J.R.; Jelkmann, W.; Schoen, C.D.

    2004-01-01

    A partial sequence of the putative polymerase (L) protein of Strawberry crinkle virus (SCV), genus Cytorhabdovirus, is described. The virus protein was found to be distantly related to the L protein of the rhabdoviruses Northern cereal mosaic virus, Rice yellow stunt virus and Sonchus yellow net

  19. Low frequency of human papillomavirus detection in prostate tissue from individuals from Northern Brazil

    Directory of Open Access Journals (Sweden)

    Rodrigo Vellasco Duarte Silvestre

    2009-07-01

    Full Text Available The presence of human papillomavirus (HPV was evaluated in 65 samples of prostate tumours and six samples of prostates with benign prostatic hyperplasia from individuals from Northern Brazil. We used a highly sensitive test, the Linear Array HPV Genotyping Test, to detect 37 high and low-risk HPV types. In this study, only 3% of tumour samples showed HPV infection. Our findings support the conclusion that, despite the high incidence of HPV infection in the geographic regions studied, HPV was not associated with a higher risk of prostate cancer. To our knowledge, this is the first study evaluating the frequency of HPV detection in prostatic tissue of individuals from Brazil.

  20. In house ELISA based on recombinant ORF2 protein underline high prevalence of IgG anti-hepatitis E virus amongst blood donors in south Brazil

    Science.gov (United States)

    Pandolfi, Rafael; Ramos de Almeida, Denise; Alves Pinto, Marcelo; Kreutz, Luiz Carlos

    2017-01-01

    Hepatitis E Virus (HEV) is a zoonotic pathogen responsible for causing acute hepatitis in human, especially in developing countries. Diagnosis of HEV usually relies on the detection of antibodies mostly by enzyme-linked immunosorbent assay (ELISA). In the present study, we designed a new indirect ELISA (iELISA) based on a short recombinant peptide derived from the capsid protein (ORF2p) and demonstrated its potential for detecting human IgG against HEV genotype 3. The best polystyrene plate (Maxisorp®), optimal ORF2p coating antigen concentration (0,67μg/well) and primary antibody dilution (1:100) were determined. This iELISA showed a sensitivity of 91.4% and specificity of 95.9%. The comparison of our in house iELISA with a commercial assay (RecomWell, Mikrogen®) showed 94.25% of agreement and a kappa index of 0.88. The ORF2 recombinant ELISA was used to screen 780 blood donors for anti-HEV IgG and we found that 314 (40,25%) of these donors were IgG positive. This high prevalence of antibodies suggests, for the first time, that the Southern Brazil region might be endemic to Hepatitis E Virus genotype 3. PMID:28486512

  1. Rapid detection of hendra virus using magnetic particles and quantum dots.

    Science.gov (United States)

    Lisi, Fabio; Falcaro, Paolo; Buso, Dario; Hill, Anita J; Barr, Jennifer A; Crameri, Gary; Nguyen, Tich-Lam; Wang, Lin-Fa; Mulvaney, Paul

    2012-09-01

    A proof-of-concept for the development of a fast and portable Hendra virus biosensor is presented. Hendra virus, a deadly emerging pathogen in Australia, can be co-localized, concentrated and revealed using simultaneously magnetic and luminescent functional particles. This method should be applicable for the early detection of any other virus by targeting the specific virus with the corresponding antibody. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. Microcephaly epidemic related to the Zika virus and living conditions in Recife, Northeast Brazil.

    Science.gov (United States)

    Souza, Wayner Vieira de; Albuquerque, Maria de Fátima Pessoa Militão de; Vazquez, Enrique; Bezerra, Luciana Caroline Albuquerque; Mendes, Antonio da Cruz Gouveia; Lyra, Tereza Maciel; Araujo, Thalia Velho Barreto de; Oliveira, André Luiz Sá de; Braga, Maria Cynthia; Ximenes, Ricardo Arraes de Alencar; Miranda-Filho, Demócrito de Barros; Cabral Silva, Amanda Priscila de Santana; Rodrigues, Laura; Martelli, Celina Maria Turchi

    2018-01-12

    Starting in August 2015, there was an increase in the number of cases of neonatal microcephaly in Northeast Brazil. These findings were identified as being an epidemic of microcephaly related to Zika virus (ZIKV) infection. The present study aims to analyse the spatial distribution of microcephaly cases in Recife (2015-2016), which is in Northeast Brazil, and its association with the living conditions in this city. This was an ecological study that used data from reported cases of microcephaly from the State Health Department of Pernambuco (August 2015 to July 2016). The basic spatial unit of analysis was the 94 districts of Recife. The case definition of microcephaly was: neonates with a head circumference of less than the cut-off point of -2 standard deviations below the mean value from the established Fenton growth curve. As an indicator of the living conditions of the 94 districts, the percentage of heads of households with an income of less than twice the minimum wage was calculated. The districts were classified into four homogeneous strata using the K-means clustering algorithm. We plotted the locations of each microcephaly case over a layer of living conditions. During the study period, 347 microcephaly cases were reported, of which 142 (40.9%) fulfilled the definition of a microcephaly case. Stratification of the 94 districts resulted in the identification of four strata. The highest stratum in relation to the living conditions presented the lowest prevalence rate of microcephaly, and the overall difference between this rate and the rates of the other strata was statistically significant. The results of the Kruskal-Wallis test demonstrated that there was a strong association between a higher prevalence of microcephaly and poor living conditions. After the first 6 months of the study period, there were no microcephaly cases recorded within the population living in the richest socio-economic strata. This study showed that those residing in areas with

  3. Detection of viruses and atypical bacteria associated with acute respiratory infection of children in Hubei, China.

    Science.gov (United States)

    Wu, Zegang; Li, Yan; Gu, Jian; Zheng, Hongyun; Tong, Yongqing; Wu, Qing

    2014-02-01

    Acute respiratory infection is the major cause of disease and death in children, particularly in developing countries. However, the spectrum of pathogenic viruses and atypical bacteria that exist in many of these countries remains incompletely characterized. The aim of this study was to examine the spectrum of pathogenic viruses and atypical bacteria associated with acute respiratory infection in children under the age of 16. A total of 10 435 serum sera specimens were collected from hospitalized children presenting with acute respiratory infection symptoms. Indirect immunofluorescence assays were performed to detect immunoglobulin M antibodies against nine common pathogens: mycoplasma pneumonia, influenza virus B, respiratory syncytial virus, parainfluenza virus, adenovirus, influenza virus A, legionella pneumophila, coxiella burnetii and chamydophila pneumonia. Of the 10 435 specimens examined, 7046 tested positive for at least one pathogen. Among all of the tested pathogens, mycoplasma pneumonia had the highest detection rate (56.9%). Influenza virus A and influenza virus B epidemics occurred during both winter and summer. The detection rate of respiratory syncytial virus and adenovirus was higher in spring. Cases of mixed infection were more complex: 4136 specimens (39.6%) tested positive for ≥2 pathogens. There were statistically significant difference in detection rates of mycoplasma pneumonia, influenza virus B, respiratory syncytial virus, parainfluenza virus, adenovirus, influenza virus A, legionella pneumophila and chamydophila pneumonia among different age groups (P acute respiratory infection among children in Hubei of China were mycoplasma pneumonia, influenza virus B and respiratory syncytial virus. The detection rates for each pathogen displayed specific seasonal and age group variations. © 2013 The Authors. Respirology © 2013 Asian Pacific Society of Respirology.

  4. Rapid Detection of the Varicella Zoster Virus in Saliva

    Science.gov (United States)

    Pierson, Duane L.; Mehta, Satish K.; Cohrs, Randall J.; Gilden, Don H.; Harding, Robert E.

    2011-01-01

    Varicella zoster virus (VZV) causes chicken pox on first exposure (usually in children), and reactivates from latency causing shingles (usually in adults). Shingles can be extremely painful, causing nerve damage, organ damage, and blindness in some cases. The virus can be life-threatening in immune-compromised individuals. The virus is very difficult to culture for diagnosis, requiring a week or longer. This invention is a rapid test for VZV from a saliva sample and can be performed in a doctor s office. The kit is small, compact, and lightweight. Detec tion is sensitive, specific, and noninvasive (no needles); only a saliva sample is required. The test provides results in minutes. The entire test is performed in a closed system, with no exposure to infectious materials. The components are made mostly of inexpensive plastic injection molded parts, many of which can be purchased off the shelf and merely assembled. All biological waste is contained for fast, efficient disposal. This innovation was made possible because of discovery of a NASA scientists flight experiment showing the presence of VZV in saliva during high stress periods and disease. This finding enables clinicians to quickly screen patients for VZV and treat the ones that show positive results with antiviral medicines. This promotes a rapid recovery, easing of pain and symptoms, and reduces chances of complications from zoster. Screening of high-risk patients could be incorporated as part of a regular physical exam. These patients include the elderly, pregnant women, and immune-compromised individuals. In these patients, VZV can be a life-threatening disease. In both high- and low-risk patients, early detection and treatment with antiviral drugs can dramatically decrease or even eliminate the clinical manifestation of disease.

  5. Detection and characterization of hepatitis A virus circulating in Egypt.

    Science.gov (United States)

    Hamza, Hazem; Abd-Elshafy, Dina Nadeem; Fayed, Sayed A; Bahgat, Mahmoud Mohamed; El-Esnawy, Nagwa Abass; Abdel-Mobdy, Emam

    2017-07-01

    Hepatitis A virus (HAV) still poses a considerable problem worldwide. In the current study, hepatitis A virus was recovered from wastewater samples collected from three wastewater treatment plants over one year. Using RT-PCR, HAV was detected in 43 out of 68 samples (63.2%) representing both inlet and outlet. Eleven positive samples were subjected to sequencing targeting the VP1-2A junction region. Phylogenetic analysis revealed that all samples belonged to subgenotype IB with few substitutions at the amino acid level. The complete sequence of one isolate (HAV/Egy/BI-11/2015) showed that the similarity at the amino acid level was not reflected at the nucleotide level. However, the deduced amino acid sequence derived from the complete nucleotide sequence showed distinct substitutions in the 2B, 2C, and 3A regions. Recombination analysis revealed a recombination event between X75215 (subgenotype IA) and AF268396 (subgenotype IB) involving a portion of the 2B nonstructural protein coding region (nucleotides 3757-3868) assuming the herein characterized sequence an actual recombinant. Despite the role of recombination in picornaviruses evolution, its involvement in HAV evolution has rarely been reported, and this may be due to the limited available complete HAV sequences. To our knowledge, this represents the first characterized complete sequence of an Egyptian isolate and the described recombination event provides an important update on the circulating HAV strains in Egypt.

  6. A molecular epidemiological study of respiratory viruses detected in Japanese children with acute wheezing illness

    Directory of Open Access Journals (Sweden)

    Noda Masahiro

    2011-06-01

    Full Text Available Abstract Background Recent studies strongly suggest that some respiratory viruses are associated with the induction of acute wheezing and/or exacerbation of bronchial asthma. However, molecular epidemiology of these viruses is not exactly known. Methods Using PCR technology, we attempted to detect various respiratory viruses from 115 Japanese children. Furthermore, the detected viruses were subjected to homology, pairwise distance, and phylogenetic analysis. Results Viruses were detected from 99 (86.1% patients. Respiratory syncytial virus (RSV alone and human rhinovirus (HRV alone were detected in 47 (40.9% and 36 (31.3% patients, respectively. Both RSV and HRV were detected in 14 (12.2% patients. Human metapneumovirus (HMPV alone and human parainfluenza virus (HPIV alone were detected in 1 (0.9% patient each, respectively. Homology and phylogenetic analyses showed that the RSV and HRV strains were classified into genetically diverse species or subgroups. In addition, RSV was the dominant virus detected in patients with no history of wheezing, whereas HRV was dominant in patients with a history of wheezing. Conclusions The results suggested that these genetically diverse respiratory viruses, especially RSV and HRV, might be associated with wheezing in Japanese children.

  7. Detection of Toxoplasma gondii DNA in fresh and frozen semen from rams in Brazil.

    Science.gov (United States)

    Bezerra, M J G; Cruz, J A L O; Kung, E S; Albuquerque, P P F; Kim, P C P; Moraes, E P B X; Pinheiro Júnior, J W; Mota, R A

    2014-10-01

    The aim of this study was to estimate the prevalence of genomic DNA of Toxoplasma gondii in semen samples from commercial rams in artificial insemination centres in Brazil, as well as in fresh semen from rams in the northeast of Brazil. In total, 108 semen samples were obtained from artificial insemination centres, and genomic DNA of T. gondii was detected in 24 of 108 (22.2%). The prevalence of antibodies anti-Toxoplasma gondii among sheep on rural properties was 9.2% (10/109), and 100% of the semen samples of these animals were positive in the PCR for T. gondii DNA. The molecular identity was confirmed through sequencing, which indicated 99.9% similarity with the T. gondii DNA sequences stored in the GenBank. This study reports the first occurrence of T. gondii DNA in the semen of rams, which came from artificial insemination centres in Brazil, as well as the occurrence of T. gondii DNA in the fresh semen of naturally infected rams in the northeast of Brazil. © 2014 Blackwell Verlag GmbH.

  8. Canine distemper virus detection in asymptomatic and non vaccinated dogs

    Directory of Open Access Journals (Sweden)

    Helen L. Del Puerto

    2010-02-01

    Full Text Available A quantitative real time polymerase chain reaction (PCR revealed canine distemper virus presence in peripheral blood samples from asymptomatic and non vaccinated dogs. Samples from eleven domestic dogs with no signs of canine distemper and not vaccinated at the month of collection were used. Canine distemper virus vaccine samples in VERO cells were used as positive controls. RNA was isolated with Trizol®, and treated with a TURBO DNA-free kit. Primers were designed for canine distemper virus nucleocapsid protein coding region fragment amplification (84 bp. Canine b-actin (93 bp was utilized as the endogenous control for normalization. Quantitative results of real time PCR generated by ABI Prism 7000 SDS Software showed that 54.5% of dogs with asymptomatic canine distemper were positive for canine distemper virus. Dissociation curves confirmed the specificity of the real time PCR fragments. This technique could detect even a few copies of viral RNA and identificate subclinically infected dogs providing accurate diagnosis of this disease at an early stage.A reação em cadeia da polimerase (PCR em tempo real revelou a presença do vírus da cinomose canina em amostra de sangue de cães assintomáticos e não vacinados. Amostra de onze cães domésticos sem nenhum sinal clínico de cinomose e que não foram vacinados no mês da coleta de sangue foram utilizados para análise. Amostra vacinal do vírus da cinomose canina em células VERO foi utilizada como controle positivo. O RNA total foi isolado utilizando-se Trizol®, e tratadas com o Kit TURBO DNA-free. Os iniciadores foram desenhados para amplificar a região do nucleocapsídeo viral com 319pb e 84pb para a PCR convencional e PCR em tempo real, respectivamente. O fragmento alvo da b-actina canina com 93pb foi utilizado como controle endógeno e normalizador. Resultados quantitativos da PCR em tempo real gerados pelo programa ABI Prism 7000 SDS demonstraram que 54,5% dos cães assintom

  9. SURVEILLANCE FOR NEWCASTLE DISEASE VIRUS, AVIAN INFLUENZA VIRUS AND MYCOPLASMA GALLISEPTICUM IN WILD BIRDS NEAR COMMERCIAL POULTRY FARMS SURROUNDED BY ATLANTIC RAINFOREST REMNANTS, SOUTHEASTERN BRAZIL

    Directory of Open Access Journals (Sweden)

    MB Guimarães

    Full Text Available ABSTRACT The geographic overlap between areas of Atlantic rainforest and human activities allows interactions to occur between humans and wild and domestic animals. Despite the great importance of the domestic animal-wildlife-human interface that occurs at poultry farms in terms of public health, economic production and wildlife conservation, there are few studies in Brazil examining the distribution and health of wild birds that interact with poultry farms. From January to December 2010, mist nets were used to capture 166 free-ranging birds that were within close proximity to three poultry farms in Atlantic rainforest remnants in south-eastern Brazil. The species composition was examined, and molecular methods were used to test for avian influenza virus, Newcastle disease virus, and Mycoplasma gallisepticum. The avian communities near the poultry farms were dominated by three synanthropic species, which corresponded to 70% of all captured individuals: house sparrows Passer domesticus (33%, saffron finches (Sicalis flaveola (22%, and ruddy ground-doves (Columbina talpacoti (15%. These predominant bird species were in poor body condition (27%, were infested with feather mites (43%, or presented both conditions (23%. No evidence of infection by avian influenza virus, Newcastle disease virus or M. gallisepticum was identified in any of the studied birds. Although no evidence of the studied pathogens was, our findings demonstrate that differences in the environmental characteristics and biosecurity practices influence the wild bird community near poultry farms, which in turn may affect the health status of these synanthropic birds and strengthen their role in the transmission of pathogens.

  10. Detection of pseudorabies virus by duplex droplet digital PCR assay.

    Science.gov (United States)

    Ren, Meishen; Lin, Hua; Chen, Shijie; Yang, Miao; An, Wei; Wang, Yin; Xue, Changhua; Sun, Yinjie; Yan, Yubao; Hu, Juan

    2018-01-01

    Aujeszky's disease, caused by pseudorabies virus (PRV), has damaged the economy of the Chinese swine industry. A large number of PRV gene-deleted vaccines have been constructed based on deletion of the glycoprotein E ( gE) gene combined with other virulence-related gene deletions, such as thymidine kinase ( TK), whereas PRV wild-type strains contain an intact gE gene. We developed a sensitive duplex droplet digital PCR (ddPCR) assay to rapidly detect PRV wild-type isolates and gE gene-deleted viral vaccines. We compared this assay with a TaqMan real-time PCR (qPCR) using the same primers and probes. Both assays exhibited good linearity and repeatability; however, ddPCR maintained linearity at extremely low concentrations, whereas qPCR did not. Based on positive results for both gE and gB, the detection limit of ddPCR was found to be 4.75 copies/µL in contrast of 76 copies/µL for qPCR, showing that ddPCR provided a 16-fold improvement in sensitivity. In addition, no nonspecific amplification was shown in specificity testing, and the PRV wild-type was distinguished from a gE-deleted strain. The ddPCR was more sensitive when analyzing clinical serum samples. Thus, ddPCR may become an appropriate detection platform for PRV.

  11. Field-Usable Lateral Flow Immunoassay for the Rapid Detection of White Spot Syndrome Virus (WSSV)

    OpenAIRE

    Kulabhusan, Prabir Kumar; Rajwade, Jyutika M.; Sugumar, Vimal; Taju, Gani; Sahul Hameed, A. S.; Paknikar, Kishore M.

    2017-01-01

    Background White spot disease (WSD), a major threat to sustainable aquaculture worldwide, is caused by White spot syndrome virus (WSSV). The diagnosis of WSD relies heavily on molecular detection of the virus by one-step PCR. These procedures are neither field-usable nor rapid enough considering the speed at which the virus spreads. Thus, development of a rapid, reliable and field-usable diagnostic method for the detection of WSSV infection is imperative to prevent huge economic losses. Metho...

  12. First detection of kobuvirus in farm animals in Brazil and the Netherlands.

    NARCIS (Netherlands)

    Barry, A.F.; Ribeiro, J.; Alfieri, A.F.; Poel, van der W.H.M.; Alfieri, A.A.

    2011-01-01

    Animal kobuviruses have been described in pigs, cattle, sheep and bats in countries in Asia and Europe. The virus can be detected in fecal and serum samples of infected animals with or without diarrhea, but most of the clinical as well as epidemiological features of kobuvirus infection are still

  13. Multi-gene detection and identification of mosquito-borne RNA viruses using an oligonucleotide microarray.

    Directory of Open Access Journals (Sweden)

    Nathan D Grubaugh

    Full Text Available BACKGROUND: Arthropod-borne viruses are important emerging pathogens world-wide. Viruses transmitted by mosquitoes, such as dengue, yellow fever, and Japanese encephalitis viruses, infect hundreds of millions of people and animals each year. Global surveillance of these viruses in mosquito vectors using molecular based assays is critical for prevention and control of the associated diseases. Here, we report an oligonucleotide DNA microarray design, termed ArboChip5.1, for multi-gene detection and identification of mosquito-borne RNA viruses from the genera Flavivirus (family Flaviviridae, Alphavirus (Togaviridae, Orthobunyavirus (Bunyaviridae, and Phlebovirus (Bunyaviridae. METHODOLOGY/PRINCIPAL FINDINGS: The assay utilizes targeted PCR amplification of three genes from each virus genus for electrochemical detection on a portable, field-tested microarray platform. Fifty-two viruses propagated in cell-culture were used to evaluate the specificity of the PCR primer sets and the ArboChip5.1 microarray capture probes. The microarray detected all of the tested viruses and differentiated between many closely related viruses such as members of the dengue, Japanese encephalitis, and Semliki Forest virus clades. Laboratory infected mosquitoes were used to simulate field samples and to determine the limits of detection. Additionally, we identified dengue virus type 3, Japanese encephalitis virus, Tembusu virus, Culex flavivirus, and a Quang Binh-like virus from mosquitoes collected in Thailand in 2011 and 2012. CONCLUSIONS/SIGNIFICANCE: We demonstrated that the described assay can be utilized in a comprehensive field surveillance program by the broad-range amplification and specific identification of arboviruses from infected mosquitoes. Furthermore, the microarray platform can be deployed in the field and viral RNA extraction to data analysis can occur in as little as 12 h. The information derived from the ArboChip5.1 microarray can help to establish

  14. A review on emerging diagnostic assay for viral detection: the case of avian influenza virus.

    NARCIS (Netherlands)

    Roodbar Shojaei, T.; Tabatabaei, M.; Shawky, S.; Salleh, M.A.M.; Bald, D.

    2015-01-01

    Biotechnology-based detection systems and sensors are in use for a wide range of applications in biomedicine, including the diagnostics of viral pathogens. In this review, emerging detection systems and their applicability for diagnostics of viruses, exemplified by the case of avian influenza virus,

  15. Detection of intestinal parasites on field-grown strawberries in the Federal District of Brazil

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    Sandra Regina Morais da Silva

    2014-12-01

    Full Text Available Introduction This study evaluated the presence of pathogenic human parasites on field-grown strawberries in the Federal District of Brazil. Methods A total of 48 samples of strawberries and 48 soil samples from 16 properties were analyzed. Results Contaminated strawberries were detected in 56% of the properties. Schistosoma mansoni, Ascaris lumbricoides or Ascaris suum, Balantidium coli, Endolimax nana, and Entamoeba spp. were detected. Soil was contaminated with Entamoeba spp., Entamoeba coli, Strongyloides spp., Ancylostomatidae, and Hymenolepis nana. Conclusions Producers should be instructed on the safe handling of strawberries in order to reduce the incidence of strawberries that are contaminated with enteroparasites.

  16. Evidence for the co-circulation of dengue virus type 3 genotypes III and V in the Northern region of Brazil during the 2002-2004 epidemics

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    Meri Bordignon Nogueira

    2008-08-01

    Full Text Available The reintroduction of dengue virus type 3 (DENV-3 in Brazil in 2000 and its subsequent spread throughout the country was associated with genotype III viruses, the only DENV-3 genotype isolated in Brazil prior to 2002. We report here the co-circulation of two different DENV-3 genotypes in patients living in the Northern region of Brazil during the 2002-2004 epidemics. Complete genomic sequences of viral RNA were determined from these epidemics, and viruses belonging to genotypes V (Southeast Asia/South Pacific and III were identified. This recent co-circulation of different DENV-3 genotypes in South America may have implications for pathological and epidemiological dynamics.

  17. High rates of detection of respiratory viruses in tonsillar tissues from children with chronic adenotonsillar disease.

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    Jose Luiz Proenca-Modena

    Full Text Available Chronic tonsillar diseases are an important health problem, leading to large numbers of surgical procedures worldwide. Little is known about pathogenesis of these diseases. In order to investigate the role of respiratory viruses in chronic adenotonsillar diseases, we developed a cross-sectional study to determine the rates of viral detections of common respiratory viruses detected by TaqMan real time PCR (qPCR in nasopharyngeal secretions, tonsillar tissues and peripheral blood from 121 children with chronic tonsillar diseases, without symptoms of acute respiratory infections. At least one respiratory virus was detected in 97.5% of patients. The viral co-infection rate was 69.5%. The most frequently detected viruses were human adenovirus in 47.1%, human enterovirus in 40.5%, human rhinovirus in 38%, human bocavirus in 29.8%, human metapneumovirus in 17.4% and human respiratory syncytial virus in 15.7%. Results of qPCR varied widely between sample sites: human adenovirus, human bocavirus and human enterovirus were predominantly detected in tissues, while human rhinovirus was more frequently detected in secretions. Rates of virus detection were remarkably high in tonsil tissues: over 85% in adenoids and close to 70% in palatine tonsils. In addition, overall virus detection rates were higher in more hypertrophic than in smaller adenoids (p = 0.05, and in the particular case of human enteroviruses, they were detected more frequently (p = 0.05 in larger palatine tonsils than in smaller ones. While persistence/latency of DNA viruses in tonsillar tissues has been documented, such is not the case of RNA viruses. Respiratory viruses are highly prevalent in adenoids and palatine tonsils of patients with chronic tonsillar diseases, and persistence of these viruses in tonsils may stimulate chronic inflammation and play a role in the pathogenesis of these diseases.

  18. High rates of detection of respiratory viruses in tonsillar tissues from children with chronic adenotonsillar disease.

    Science.gov (United States)

    Proenca-Modena, Jose Luiz; Pereira Valera, Fabiana Cardoso; Jacob, Marcos Gerhardinger; Buzatto, Guilherme Pietrucci; Saturno, Tamara Honorato; Lopes, Lucia; Souza, Jamila Mendonça; Escremim Paula, Flavia; Silva, Maria Lucia; Carenzi, Lucas Rodrigues; Tamashiro, Edwin; Arruda, Eurico; Anselmo-Lima, Wilma Terezinha

    2012-01-01

    Chronic tonsillar diseases are an important health problem, leading to large numbers of surgical procedures worldwide. Little is known about pathogenesis of these diseases. In order to investigate the role of respiratory viruses in chronic adenotonsillar diseases, we developed a cross-sectional study to determine the rates of viral detections of common respiratory viruses detected by TaqMan real time PCR (qPCR) in nasopharyngeal secretions, tonsillar tissues and peripheral blood from 121 children with chronic tonsillar diseases, without symptoms of acute respiratory infections. At least one respiratory virus was detected in 97.5% of patients. The viral co-infection rate was 69.5%. The most frequently detected viruses were human adenovirus in 47.1%, human enterovirus in 40.5%, human rhinovirus in 38%, human bocavirus in 29.8%, human metapneumovirus in 17.4% and human respiratory syncytial virus in 15.7%. Results of qPCR varied widely between sample sites: human adenovirus, human bocavirus and human enterovirus were predominantly detected in tissues, while human rhinovirus was more frequently detected in secretions. Rates of virus detection were remarkably high in tonsil tissues: over 85% in adenoids and close to 70% in palatine tonsils. In addition, overall virus detection rates were higher in more hypertrophic than in smaller adenoids (p = 0.05), and in the particular case of human enteroviruses, they were detected more frequently (p = 0.05) in larger palatine tonsils than in smaller ones. While persistence/latency of DNA viruses in tonsillar tissues has been documented, such is not the case of RNA viruses. Respiratory viruses are highly prevalent in adenoids and palatine tonsils of patients with chronic tonsillar diseases, and persistence of these viruses in tonsils may stimulate chronic inflammation and play a role in the pathogenesis of these diseases.

  19. Identify-Isolate-Inform: A Tool for Initial Detection and Management of Zika Virus Patients in the Emergency Department

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    Kristi L. Koenig

    2016-05-01

    Full Text Available First isolated in 1947 from a monkey in the Zika forest in Uganda, and from mosquitoes in the same forest the following year, Zika virus has gained international attention due to concerns for infection in pregnant women potentially causing fetal microcephaly. More than one million people have been infected since the appearance of the virus in Brazil in 2015. Approximately 80% of infected patients are asymptomatic. An association with microcephaly and other birth defects as well as Guillain-Barre Syndrome has led to a World Health Organization declaration of Zika virus as a Public Health Emergency of International Concern in February 2016. Zika virus is a vector-borne disease transmitted primarily by the Aedes aegypti mosquito. Male to female sexual transmission has been reported and there is potential for transmission via blood transfusions. After an incubation period of 2-7 days, symptomatic patients develop rapid onset fever, maculopapular rash, arthralgia, and conjunctivitis, often associated with headache and myalgias. Emergency department (ED personnel must be prepared to address concerns from patients presenting with symptoms consistent with acute Zika virus infection, especially those who are pregnant or planning travel to Zika-endemic regions, as well as those women planning to become pregnant and their partners. The identify-isolate-inform (3I tool, originally conceived for initial detection and management of Ebola virus disease patients in the ED, and later adjusted for measles and Middle East Respiratory Syndrome, can be adapted for real-time use for any emerging infectious disease. This paper reports a modification of the 3I tool for initial detection and management of patients under investigation for Zika virus. Following an assessment of epidemiologic risk, including travel to countries with mosquitoes that transmit Zika virus, patients are further investigated if clinically indicated. If after a rapid evaluation, Zika or other

  20. Identify-Isolate-Inform: A Tool for Initial Detection and Management of Zika Virus Patients in the Emergency Department.

    Science.gov (United States)

    Koenig, Kristi L; Almadhyan, Abdulmajeed; Burns, Michael J

    2016-05-01

    First isolated in 1947 from a monkey in the Zika forest in Uganda, and from mosquitoes in the same forest the following year, Zika virus has gained international attention due to concerns for infection in pregnant women potentially causing fetal microcephaly. More than one million people have been infected since the appearance of the virus in Brazil in 2015. Approximately 80% of infected patients are asymptomatic. An association with microcephaly and other birth defects as well as Guillain-Barre Syndrome has led to a World Health Organization declaration of Zika virus as a Public Health Emergency of International Concern in February 2016. Zika virus is a vector-borne disease transmitted primarily by the Aedes aegypti mosquito. Male to female sexual transmission has been reported and there is potential for transmission via blood transfusions. After an incubation period of 2-7 days, symptomatic patients develop rapid onset fever, maculopapular rash, arthralgia, and conjunctivitis, often associated with headache and myalgias. Emergency department (ED) personnel must be prepared to address concerns from patients presenting with symptoms consistent with acute Zika virus infection, especially those who are pregnant or planning travel to Zika-endemic regions, as well as those women planning to become pregnant and their partners. The identify-isolate-inform (3I) tool, originally conceived for initial detection and management of Ebola virus disease patients in the ED, and later adjusted for measles and Middle East Respiratory Syndrome, can be adapted for real-time use for any emerging infectious disease. This paper reports a modification of the 3I tool for initial detection and management of patients under investigation for Zika virus. Following an assessment of epidemiologic risk, including travel to countries with mosquitoes that transmit Zika virus, patients are further investigated if clinically indicated. If after a rapid evaluation, Zika or other arthropod

  1. Nested PCR for Rapid Detection of Mumps Virus in Cerebrospinal Fluid from Patients with Neurological Diseases

    Science.gov (United States)

    Poggio, Gustavo Palacios; Rodriguez, Claudia; Cisterna, Daniel; Freire, María Cecilia; Cello, Jerónimo

    2000-01-01

    In this study, we have developed a reverse transcription (RT)-nested polymerase chain reaction (n-PCR) for the detection of mumps virus RNA in cerebrospinal fluid (CSF) from patients with neurological infections. A specific 112-bp fragment was amplified by this method with primers from the nucleoprotein of the mumps virus genome. The mumps virus RT–n-PCR was capable of detecting 0.001 PFU/ml and 0.005 50% tissue culture infective dose/ml. This method was found to be specific, since no PCR product was detected in each of the CSF samples from patients with proven non-mumps virus-related meningitis or encephalitis. Mumps virus RNA was detected in all 18 CSF samples confirmed by culture to be infected with mumps virus. Positive PCR results were obtained for the CSF of 26 of 28 patients that were positive for signs of mumps virus infection (i.e., cultivable virus from urine or oropharyngeal samples or positivity for anti-mumps virus immunoglobulin M) but without cultivable virus in their CSF. Overall, mumps virus RNA was detected in CSF of 96% of the patients with a clinical diagnosis of viral central nervous system (CNS) disease and confirmed mumps virus infection, while mumps virus was isolated in CSF of only 39% of the patients. Furthermore, in a retrospective study, we were able to detect mumps virus RNA in 25 of 55 (46%) CSF samples from patients with a clinical diagnosis of viral CNS disease and negative laboratory evidence of viral infection including mumps virus infection. The 25 patients represent 12% of the 236 patients who had a clinical diagnosis of viral CNS infections and whose CSF was examined at our laboratory for a 2-year period. The findings confirm the importance of mumps virus as a causative agent of CNS infections in countries with low vaccine coverage rates. In summary, our study demonstrates the usefulness of the mumps virus RT–n-PCR for the diagnosis of mumps virus CNS disease and suggests that this assay may soon become the “gold standard

  2. Rabies Virus in Bats, State of Pará, Brazil, 2005-2011.

    Science.gov (United States)

    Pereira, Armando de Souza; Casseb, Livia Medeiros Neves; Barbosa, Taciana Fernandes Souza; Begot, Alberto Lopes; Brito, Roberto Messias Oliveira; Vasconcelos, Pedro Fernando da Costa; Travassos da Rosa, Elizabeth Salbé

    2017-08-01

    Rabies is an acute, progressive zoonotic viral infection that in general produces a fatal outcome. This disease is responsible for deaths in humans and animals worldwide and, because it can affect all mammals, is considered one of the most important viral infections for public health. This study aimed to determine the prevalence of rabies in bats of different species found in municipalities of the state of Pará from 2005 to 2011. The rabies virus was detected in 12 (0.39%) bats in a total of 3100 analyzed, including hematophagous, frugivorous, and insectivorous bats. Of these, eleven were characterized as AgV3, which is characteristic of the hematophagous bat Desmodus rotundus (E. Geoffroy 1810); one insectivorous animal showed a different profile compatible with the Eptesicus pattern and may therefore be a new antigenic variant. This study identified the need for greater intensification of epidemiological surveillance in municipalities lacking rabies surveillance (silent areas); studies of rabies virus in bats with different alimentary habits, studies investigating the prevalence of AgV3, and prophylactic measures in areas where humans may be infected are also needed.

  3. Multiplex RT-PCR method for the simultaneous detection of nine grapevine viruses.

    Science.gov (United States)

    Gambino, Giorgio

    2015-01-01

    Viral diseases are a serious pathological problem for grapevines, and in recent years the need for increasingly specific and rapid diagnostic methods for the selection of propagation materials has grown. Arabis mosaic virus, Grapevine fanleaf virus, Grapevine virus A, Grapevine virus B, Grapevine rupestris stem pitting-associated virus, Grapevine fleck virus, and Grapevine leafroll-associated viruses 1, 2, and 3 are nine of the most widespread viruses that naturally infect grapevines. A multiplex RT-PCR was developed for simultaneous detection of these nine grapevine viruses, in combination with a plant RNA internal control used as an indicator of the effectiveness of the reaction. One to ten fragments specific for the viruses and an internal control were simultaneously amplified from infected samples and identified by their specific molecular sizes in agarose gel. The protocol reported is an update of previously published protocols for RNA extraction and multiplex diagnosis of viruses. After several years of use and hundreds of samples tested, and following validation in several laboratories, this multiplex RT-PCR provides a reliable and rapid method for detecting grapevine viruses from a large number of samples.

  4. Highly Specific Detection of Five Exotic Quarantine Plant Viruses using RT-PCR.

    Science.gov (United States)

    Choi, Hoseong; Cho, Won Kyong; Yu, Jisuk; Lee, Jong-Seung; Kim, Kook-Hyung

    2013-03-01

    To detect five plant viruses (Beet black scorch virus, Beet necrotic yellow vein virus, Eggplant mottled dwarf virus, Pelargonium zonate spot virus, and Rice yellow mottle virus) for quarantine purposes, we designed 15 RT-PCR primer sets. Primer design was based on the nucleotide sequence of the coat protein gene, which is highly conserved within species. All but one primer set successfully amplified the targets, and gradient PCRs indicated that the optimal temperature for the 14 useful primer sets was 51.9°C. Some primer sets worked well regardless of annealing temperature while others required a very specific annealing temperature. A primer specificity test using plant total RNAs and cDNAs of other plant virus-infected samples demonstrated that the designed primer sets were highly specific and generated reproducible results. The newly developed RT-PCR primer sets would be useful for quarantine inspections aimed at preventing the entry of exotic plant viruses into Korea.

  5. Persistence of Zika Virus in Breast Milk after Infection in Late Stage of Pregnancy

    Science.gov (United States)

    Sotelo, José R.; Sotelo, Andre B.; Sotelo, Fabio J.B.; Pinho, Joao R.R.; Oliveira, Rita de Cassia; Bezerra, Alanna M.P.S.; Deutsch, Alice D.; Villas-Boas, Lucy S.; Felix, Alvina C.; Romano, Camila M.; Machado, Clarisse M.; Mendes-Correa, Maria C.J.; Santana, Rubia A.F.; Menezes, Fernando G.; Mangueira, Cristovao L.P.

    2017-01-01

    We detected Zika virus in breast milk of a woman in Brazil infected with the virus during the 36th week of pregnancy. Virus was detected 33 days after onset of signs and symptoms and 9 days after delivery. No abnormalities were found during fetal assessment or after birth of the infant. PMID:28192072

  6. Persistence of Zika Virus in Breast Milk after Infection in Late Stage of Pregnancy.

    Science.gov (United States)

    Sotelo, José R; Sotelo, Andre B; Sotelo, Fabio J B; Doi, André M; Pinho, Joao R R; Oliveira, Rita de Cassia; Bezerra, Alanna M P S; Deutsch, Alice D; Villas-Boas, Lucy S; Felix, Alvina C; Romano, Camila M; Machado, Clarisse M; Mendes-Correa, Maria C J; Santana, Rubia A F; Menezes, Fernando G; Mangueira, Cristovao L P

    2017-05-01

    We detected Zika virus in breast milk of a woman in Brazil infected with the virus during the 36th week of pregnancy. Virus was detected 33 days after onset of signs and symptoms and 9 days after delivery. No abnormalities were found during fetal assessment or after birth of the infant.

  7. Assessment of Gastroenteric Viruses Frequency in a Children's Day Care Center in Rio De Janeiro, Brazil: A Fifteen Year Study (1994–2008)

    Science.gov (United States)

    Ferreira, Mônica Simões Rocha; Xavier, Maria da Penha Trindade Pinheiro; Tinga, Anna Carolina De Castro; Rose, Tatiana Lundgren; Fumian, Tulio Machado; Fialho, Alexandre Madi; de Assis, Rosane Maria; Costa, Filipe Aníbal Carvalho; de Oliveira, Solange Artimos; Leite, José Paulo Gagliardi; Miagostovich, Marize Pereira

    2012-01-01

    This 15-year study aimed to determine the role of the main viruses responsible for acute infantile gastroenteritis cases in a day care center in the city of Rio de Janeiro, Brazil. From 1994 to 2008, 539 fecal samples were obtained from 23 outbreaks as well as sporadic cases that occurred in this period. The detection of Rotavirus group A (RVA), norovirus (NoV) and astrovirus (AstV) was investigated both by classical and molecular methods of viral detection. RVA was detected by enzymatic immune assay and/or polyacrylamide gel electrophoresis and genotyped by using semi-nested multiplex PCR. NoV and AstV were subsequently tested by real time PCR in all RVA-negative samples and genotyped throughout genome sequencing. Three protocols for molecular characterization of NoV nucleotide sequencing were performed with the partial nucleotide sequencing of genomic regions known as region B (polymerase gen), C and D (capsid gen).Viruses were identified in 47.7% (257/539) of the cases, and the detection rates of RVA, NoV and AstV in16.1% (87/539), 33.4% (151/452), and 6.3% (19/301), respectively. Most gastroenteritis cases were reported in autumn and winter, although NoV presented a broader monthly distribution. Viruses' detection rates were significantly higher among children aged less than 24 months old, although NoV cases were detected in all age groups. RVA genotypes as G1P[8], G9P[8], G2P[4], G3P[8] and G1+G3P[8] and RVA was no longer detected after 2005. NoV characterization revealed genotypes variability circulating in the period as GI.2, GI.3, GI.8 GII.2, GII.3, GII.4, GII.4 variants 2001 and 2006b, GII.6, GII.7, GII.12 and GII.17. AstV genotypes 1, 2, 4 and 5 were also characterized. Those data demonstrate the impact of NoV infection in cases of infantile gastroenteritis, surpassing RVA infection responsible for high morbidity rate in children under five years old. PMID:22448271

  8. Assessment of gastroenteric viruses frequency in a children's day care center in Rio De Janeiro, Brazil: a fifteen year study (1994-2008.

    Directory of Open Access Journals (Sweden)

    Mônica Simões Rocha Ferreira

    Full Text Available This 15-year study aimed to determine the role of the main viruses responsible for acute infantile gastroenteritis cases in a day care center in the city of Rio de Janeiro, Brazil. From 1994 to 2008, 539 fecal samples were obtained from 23 outbreaks as well as sporadic cases that occurred in this period. The detection of Rotavirus group A (RVA, norovirus (NoV and astrovirus (AstV was investigated both by classical and molecular methods of viral detection. RVA was detected by enzymatic immune assay and/or polyacrylamide gel electrophoresis and genotyped by using semi-nested multiplex PCR. NoV and AstV were subsequently tested by real time PCR in all RVA-negative samples and genotyped throughout genome sequencing. Three protocols for molecular characterization of NoV nucleotide sequencing were performed with the partial nucleotide sequencing of genomic regions known as region B (polymerase gen, C and D (capsid gen.Viruses were identified in 47.7% (257/539 of the cases, and the detection rates of RVA, NoV and AstV in16.1% (87/539, 33.4% (151/452, and 6.3% (19/301, respectively. Most gastroenteritis cases were reported in autumn and winter, although NoV presented a broader monthly distribution. Viruses' detection rates were significantly higher among children aged less than 24 months old, although NoV cases were detected in all age groups. RVA genotypes as G1P[8], G9P[8], G2P[4], G3P[8] and G1+G3P[8] and RVA was no longer detected after 2005. NoV characterization revealed genotypes variability circulating in the period as GI.2, GI.3, GI.8 GII.2, GII.3, GII.4, GII.4 variants 2001 and 2006b, GII.6, GII.7, GII.12 and GII.17. AstV genotypes 1, 2, 4 and 5 were also characterized. Those data demonstrate the impact of NoV infection in cases of infantile gastroenteritis, surpassing RVA infection responsible for high morbidity rate in children under five years old.

  9. Detection of Leishmania RNA virus in Leishmania parasites.

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    Haroun Zangger

    Full Text Available Patients suffering from cutaneous leishmaniasis (CL caused by New World Leishmania (Viannia species are at high risk of developing mucosal (ML or disseminated cutaneous leishmaniasis (DCL. After the formation of a primary skin lesion at the site of the bite by a Leishmania-infected sand fly, the infection can disseminate to form secondary lesions. This metastatic phenotype causes significant morbidity and is often associated with a hyper-inflammatory immune response leading to the destruction of nasopharyngeal tissues in ML, and appearance of nodules or numerous ulcerated skin lesions in DCL. Recently, we connected this aggressive phenotype to the presence of Leishmania RNA virus (LRV in strains of L. guyanensis, showing that LRV is responsible for elevated parasitaemia, destructive hyper-inflammation and an overall exacerbation of the disease. Further studies of this relationship and the distribution of LRVs in other Leishmania strains and species would benefit from improved methods of viral detection and quantitation, especially ones not dependent on prior knowledge of the viral sequence as LRVs show significant evolutionary divergence.This study reports various techniques, among which, the use of an anti-dsRNA monoclonal antibody (J2 stands out for its specific and quantitative recognition of dsRNA in a sequence-independent fashion. Applications of J2 include immunofluorescence, ELISA and dot blot: techniques complementing an arsenal of other detection tools, such as nucleic acid purification and quantitative real-time-PCR. We evaluate each method as well as demonstrate a successful LRV detection by the J2 antibody in several parasite strains, a freshly isolated patient sample and lesion biopsies of infected mice.We propose that refinements of these methods could be transferred to the field for use as a diagnostic tool in detecting the presence of LRV, and potentially assessing the LRV-related risk of complications in cutaneous leishmaniasis.

  10. Viruses detected among sporadic cases of parotitis, United States, 2009-2011.

    Science.gov (United States)

    Barskey, Albert E; Juieng, Phalasy; Whitaker, Brett L; Erdman, Dean D; Oberste, M Steven; Chern, Shur-Wern Wang; Schmid, D Scott; Radford, Kay W; McNall, Rebecca J; Rota, Paul A; Hickman, Carole J; Bellini, William J; Wallace, Gregory S

    2013-12-15

    Sporadic cases of parotitis are generally assumed to be mumps, which often requires a resource-intensive public health response. This project surveyed the frequency of viruses detected among such cases. During 2009-2011, 8 jurisdictions throughout the United States investigated sporadic cases of parotitis. Epidemiologic information, serum, and buccal and oropharyngeal swabs were collected. Polymerase chain reaction methods were used to detect a panel of viruses. Anti-mumps virus immunoglobulin M (IgM) antibodies were detected using a variety of methods. Of 101 specimens, 38 were positive for a single virus: Epstein-Barr virus (23), human herpesvirus (HHV)-6B (10), human parainfluenza virus (HPIV)-2 (3), HPIV-3 (1), and human bocavirus (1). Mumps virus, enteroviruses (including human parechovirus), HHV-6A, HPIV-1, and adenoviruses were not detected. Early specimen collection did not improve viral detection rate. Mumps IgM was detected in 17% of available specimens. Patients in whom a virus was detected were younger, but no difference was seen by sex or vaccination profile. No seasonal patterns were identified. Considering the timing of specimen collection, serology results, patient vaccination status, and time of year may be helpful in assessing the likelihood that a sporadic case of parotitis without laboratory confirmation is mumps.

  11. Tracking the return of Aedes aegypti to Brazil, the major vector of the dengue, chikungunya and Zika viruses.

    Science.gov (United States)

    Kotsakiozi, Panayiota; Gloria-Soria, Andrea; Caccone, Adalgisa; Evans, Benjamin; Schama, Renata; Martins, Ademir Jesus; Powell, Jeffrey R

    2017-07-01

    Aedes aegypti, commonly known as "the yellow fever mosquito", is of great medical concern today primarily as the major vector of dengue, chikungunya and Zika viruses, although yellow fever remains a serious health concern in some regions. The history of Ae. aegypti in Brazil is of particular interest because the country was subjected to a well-documented eradication program during 1940s-1950s. After cessation of the campaign, the mosquito quickly re-established in the early 1970s with several dengue outbreaks reported during the last 30 years. Brazil can be considered the country suffering the most from the yellow fever mosquito, given the high number of dengue, chikungunya and Zika cases reported in the country, after having once been declared "free of Ae. aegypti". We used 12 microsatellite markers to infer the genetic structure of Brazilian Ae. aegypti populations, genetic variability, genetic affinities with neighboring geographic areas, and the timing of their arrival and spread. This enabled us to reconstruct their recent history and evaluate whether the reappearance in Brazil was the result of re-invasion from neighboring non-eradicated areas or re-emergence from local refugia surviving the eradication program. Our results indicate a genetic break separating the northern and southern Brazilian Ae. aegypti populations, with further genetic differentiation within each cluster, especially in southern Brazil. Based on our results, re-invasions from non-eradicated regions are the most likely scenario for the reappearance of Ae. aegypti in Brazil. While populations in the northern cluster are likely to have descended from Venezuela populations as early as the 1970s, southern populations seem to have derived more recently from northern Brazilian areas. Possible entry points are also revealed within both southern and northern clusters that could inform strategies to control and monitor this important arbovirus vector.

  12. Plant viruses in aqueous environment - survival, water mediated transmission and detection.

    Science.gov (United States)

    Mehle, Nataša; Ravnikar, Maja

    2012-10-15

    The presence of plant viruses outside their plant host or insect vectors has not been studied intensively. This is due, in part, to the lack of effective detection methods that would enable their detection in difficult matrixes and in low titres, and support the search for unknown viruses. Recently, new and sensitive methods for detecting viruses have resulted in a deeper insight into plant virus movement through, and transmission between, plants. In this review, we have focused on plant viruses found in environmental waters and their detection. Infectious plant pathogenic viruses from at least 7 different genera have been found in aqueous environment. The majority of the plant pathogenic viruses so far recovered from environmental waters are very stable, they can infect plants via the roots without the aid of a vector and often have a wide host range. The release of such viruses from plants can lead to their dissemination in streams, lakes, and rivers, thereby ensuring the long-distance spread of viruses that otherwise, under natural conditions, would remain restricted to limited areas. The possible sources and survival of plant viruses in waters are therefore discussed. Due to the widespread use of hydroponic systems and intensive irrigation in horticulture, the review is focused on the possibility and importance of spreading viral infection by water, together with measures for preventing the spread of viruses. The development of new methods for detecting multiple plant viruses at the same time, like microarrays or new generation sequencing, will facilitate the monitoring of environmental waters and waters used for irrigation and in hydroponic systems. It is reasonable to expect that the list of plant viruses found in waters will thereby be expanded considerably. This will emphasize the need for further studies to determine the biological significance of water-mediated transport. Copyright © 2012 Elsevier Ltd. All rights reserved.

  13. Multiplex hydrolysis probe real-time PCR for simultaneous detection of hepatitis A virus and hepatitis E virus.

    Science.gov (United States)

    Qiu, Feng; Cao, Jingyuan; Su, Qiudong; Yi, Yao; Bi, Shengli

    2014-05-30

    Detection of hepatitis viral infections has traditionally relied on the circulating antibody test using the enzyme-linked immunosorbent assay. However, multiplex real-time PCR has been increasingly used for a variety of viral nucleic acid detections and has proven to be superior to traditional methods. Hepatitis A virus (HAV) and hepatitis E virus (HEV) are the major causes of acute hepatitis worldwide; both HAV and HEV infection are a main public health problem. In the present study, a one-step multiplex reverse transcriptase quantitative polymerase chain reaction assay using hydrolysis probes was developed for simultaneously detecting HAV and HEV. This novel detection system proved specific to the target viruses, to be highly sensitive and to be applicable to clinical sera samples, making it useful for rapid, accurate and feasible identification of HAV and HEV.

  14. Prospective study of human herpesvirus 8 oral shedding, viremia, and serological status among human immunodeficiency virus seropositive and seronegative individuals in Sao Paulo, Brazil.

    Science.gov (United States)

    Braz-Silva, Paulo H; Tozetto-Mendoza, Tania R; Sumita, Laura M; Freire, Wilton; Palmieri, Michelle; do Canto, Alan M; Avelino-Silva, Vivian I; Gallottini, Marina; Mayaud, Philippe; Pannuti, Claudio S

    2017-01-01

    Human herpesvirus 8 (HHV-8) is a gamma-herpesvirus and etiological agent of all forms of Kaposi sarcoma (KS). Saliva may play an important role in HHV-8 transmission in specific populations. Little is known about HHV-8 oral shedding pattern and the possible correlation with the HHV-8 serological profile and viremia. A prospective study was conducted of HHV-8 salivary excretion among human immunodeficiency virus HIV-seronegative (n = 47) and -seropositive (n = 44) homosexual men and HIV-seropositive women (n = 32) over a 6-month period with monthly HHV-8 serologies (immunofluorescence assays to identify antibodies to latent and lytic HHV-8 viral proteins, and a whole-virus HHV-8 enzyme-linked immunosorbent assay [ELISA]), monthly HHV-8 DNA serum/plasma detection, and daily self-collected oral rinses for HHV-8-DNA detection using real-time polymerase chain reaction. HHV-8 seropositivity was 51.1%, 63.6%, and 37.5%, in the three studied groups. There was no case of HHV-8 DNA detection in serum/plasma. Intermittent detection of oral HHV-8 DNA was observed during 5.1% (110/2,160) of visits among 28% (18/64) of HHV-8-seropositive individuals, all of whom were males and HHV-8 ELISA seropositive. In immunologically controlled populations of Brazil, HHV-8 oral shedding was limited to HHV-8-seropositive men, occurred infrequently and intermittently, and was not linked to HHV-8 viremia, suggesting a limited potential for oral or blood transmission.

  15. DETECTION OF VIRULENCE GENES IN ENVIRONMENTAL STRAINS OF Vibrio cholerae FROM ESTUARIES IN NORTHEASTERN BRAZIL

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    Francisca Gleire Rodrigues de Menezes

    2014-09-01

    Full Text Available The objectives of this study were to detect the presence of Vibrio cholerae in tropical estuaries (Northeastern Brazil and to search for virulence factors in the environmental isolates. Water and sediment samples were inoculated onto a vibrio-selective medium (TCBS, and colonies with morphological resemblance to V. cholerae were isolated. The cultures were identified phenotypically using a dichotomous key based on biochemical characteristics. The total DNA extracted was amplified by PCR to detect ompW and by multiplex PCR to detect the virulence genes ctx, tcp, zot and rfbO1. The results of the phenotypic and genotypic identification were compared. Nine strains of V. cholerae were identified phenotypically, five of which were confirmed by detection of the species-specific gene ompW. The dichotomous key was efficient at differentiating environmental strains of V. cholerae. Strains of V. cholerae were found in all four estuaries, but none possessed virulence genes.

  16. Detection of virulence genes in environmental strains of Vibrio cholerae from estuaries in northeastern Brazil.

    Science.gov (United States)

    Menezes, Francisca Gleire Rodrigues de; Neves, Soraya da Silva; Sousa, Oscarina Viana de; Vila-Nova, Candida Machado Vieira Maia; Maggioni, Rodrigo; Theophilo, Grace Nazareth Diogo; Hofer, Ernesto; Vieira, Regine Helena Silva dos Fernandes

    2014-01-01

    The objectives of this study were to detect the presence of Vibrio cholerae in tropical estuaries (Northeastern Brazil) and to search for virulence factors in the environmental isolates. Water and sediment samples were inoculated onto a vibrio-selective medium (TCBS), and colonies with morphological resemblance to V. cholerae were isolated. The cultures were identified phenotypically using a dichotomous key based on biochemical characteristics. The total DNA extracted was amplified by PCR to detect ompW and by multiplex PCR to detect the virulence genes ctx, tcp, zot and rfbO1. The results of the phenotypic and genotypic identification were compared. Nine strains of V. cholerae were identified phenotypically, five of which were confirmed by detection of the species-specific gene ompW. The dichotomous key was efficient at differentiating environmental strains of V. cholerae. Strains of V. cholerae were found in all four estuaries, but none possessed virulence genes.

  17. DETECTION OF VIRULENCE GENES IN ENVIRONMENTAL STRAINS OF Vibrio cholerae FROM ESTUARIES IN NORTHEASTERN BRAZIL

    Science.gov (United States)

    de Menezes, Francisca Gleire Rodrigues; Neves, Soraya da Silva; de Sousa, Oscarina Viana; Vila-Nova, Candida Machado Vieira Maia; Maggioni, Rodrigo; Theophilo, Grace Nazareth Diogo; Hofer, Ernesto; Vieira, Regine Helena Silva dos Fernandes

    2014-01-01

    The objectives of this study were to detect the presence of Vibrio cholerae in tropical estuaries (Northeastern Brazil) and to search for virulence factors in the environmental isolates. Water and sediment samples were inoculated onto a vibrio-selective medium (TCBS), and colonies with morphological resemblance to V. cholerae were isolated. The cultures were identified phenotypically using a dichotomous key based on biochemical characteristics. The total DNA extracted was amplified by PCR to detect ompW and by multiplex PCR to detect the virulence genes ctx, tcp, zot and rfbO1. The results of the phenotypic and genotypic identification were compared. Nine strains of V. cholerae were identified phenotypically, five of which were confirmed by detection of the species-specific gene ompW. The dichotomous key was efficient at differentiating environmental strains of V. cholerae. Strains of V. cholerae were found in all four estuaries, but none possessed virulence genes. PMID:25229224

  18. Viruses and human cancer: from detection to causality.

    Science.gov (United States)

    Sarid, Ronit; Gao, Shou-Jiang

    2011-06-28

    The study of cancer is incomplete without taking into consideration of tumorigenic viruses. Initially, searches for human cancer viruses were fruitless despite an expansion of our knowledge in the same period concerning acute-transforming retroviruses in animals. However, over the last 40 years, we have witnessed rapid progress in the tumor virology field. Currently, acknowledged human cancer viruses include Epstein-Barr virus, hepatitis B virus, hepatitis C virus, high-risk human papilloma viruses, human T-cell lymphotropic virus type 1 and Kaposi's sarcoma-associated herpesvirus. Extensive epidemiological and mechanistic studies have led to the development of novel preventive and therapeutic approaches for managing some of these infections and associated cancers. In addition, recent advances in molecular technologies have enabled the discovery of a new potential human tumor virus, Merkel cell polyomavirus, but its association with cancer remains to be validated. It is anticipated that in the next few decades many additional human cancer viruses will be discovered and the mechanisms underlying viral oncogenesis delineated. Thus, it can be expected that better tools for preventing and treating virus-associated cancer will be available in the near future. Copyright © 2010 Elsevier Ireland Ltd. All rights reserved.

  19. A Multicenter Study To Evaluate the Performance of High-Throughput Sequencing for Virus Detection.

    Science.gov (United States)

    Khan, Arifa S; Ng, Siemon H S; Vandeputte, Olivier; Aljanahi, Aisha; Deyati, Avisek; Cassart, Jean-Pol; Charlebois, Robert L; Taliaferro, Lanyn P

    2017-01-01

    The capability of high-throughput sequencing (HTS) for detection of known and unknown viruses makes it a powerful tool for broad microbial investigations, such as evaluation of novel cell substrates that may be used for the development of new biological products. However, like any new assay, regulatory applications of HTS need method standardization. Therefore, our three laboratories initiated a study to evaluate performance of HTS for potential detection of viral adventitious agents by spiking model viruses in different cellular matrices to mimic putative materials for manufacturing of biologics. Four model viruses were selected based upon different physical and biochemical properties and commercial availability: human respiratory syncytial virus (RSV), Epstein-Barr virus (EBV), feline leukemia virus (FeLV), and human reovirus (REO). Additionally, porcine circovirus (PCV) was tested by one laboratory. Independent samples were prepared for HTS by spiking intact viruses or extracted viral nucleic acids, singly or mixed, into different HeLa cell matrices (resuspended whole cells, cell lysate, or total cellular RNA). Data were obtained using different sequencing platforms (Roche 454, Illumina HiSeq1500 or HiSeq2500). Bioinformatic analyses were performed independently by each laboratory using available tools, pipelines, and databases. The results showed that comparable virus detection was obtained in the three laboratories regardless of sample processing, library preparation, sequencing platform, and bioinformatic analysis: between 0.1 and 3 viral genome copies per cell were detected for all of the model viruses used. This study highlights the potential for using HTS for sensitive detection of adventitious viruses in complex biological samples containing cellular background. IMPORTANCE Recent high-throughput sequencing (HTS) investigations have resulted in unexpected discoveries of known and novel viruses in a variety of sample types, including research materials

  20. Sequence analysis of the capsid gene of Aichi viruses detected from Japan, Bangladesh, Thailand, and Vietnam.

    Science.gov (United States)

    Pham, Ngan Thi Kim; Trinh, Quang Duy; Khamrin, Pattara; Nguyen, Tuan Anh; Dey, Shuvra Kanti; Phan, Tung Gia; Hoang, Le Phuc; Maneekarn, Niwat; Okitsu, Shoko; Mizuguchi, Masashi; Ushijima, Hiroshi

    2008-07-01

    Sequence analysis of the capsid gene of Aichi viruses was performed on 12 strains detected in Japan, Bangladesh, Thailand, and Vietnam during 2002-2005. The phylogenetic tree constructed from 17 nucleotide sequences of the capsid gene of the strains studied and reference strains demonstrated that Aichi virus strains clustered into two branches. A classification of Aichi viruses based on the capsid gene was proposed, in which lineage I consists of the Aichi virus strains detected from Japan, Thailand, Vietnam, and Germany, and lineage II includes Bangladeshi strains and a Brazilian strain.

  1. The early use of yellow fever virus strain 17D for vaccine production in Brazil - a review

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    Paulo Roberto Post

    2001-08-01

    Full Text Available The use of yellow fever (YF virus 17D strain for vaccine production adapted in Brazil since its introduction in 1937 was reviewed. This was possible due to the availability of official records of vaccine production. The retrieved data highlight the simultaneous use of several serially passaged 17D substrain viruses for both inocula and vaccine preparation that allowed uninterrupted production. Substitution of these substrain viruses became possible with the experience gained during quality control and human vaccination. Post-vaccinal complications in humans and the failure of some viruses in quality control tests (neurovirulence for monkeys indicated that variables needed to be reduced during vaccine production, leading to the development of the seed lot system. The 17DD substrain, still used today, was the most frequently used substrain and the most reliable in terms of safety and efficacy. For this reason, it is possible to derive an infectious cDNA clone of this substrain combined with production in cell culture that could be used to direct the expression of heterologous antigens and lead to the development of new live vaccines.

  2. Detection of Mycobacterium avium subsp. paratuberculosis in bovine milk from the state of Pernambuco, Brazil.

    Science.gov (United States)

    Albuquerque, Pedro Paulo Feitosa de; Santos, André de Souza; Souza Neto, Orestes Luiz de; Kim, Pomy de Cássia Peixoto; Cavalcanti, Erika Fernanda Torres Samico Fernandes; Oliveira, Júnior Mário Baltazar de; Mota, Rinaldo Aparecido; Júnior, José Wilton Pinheiro

    The aim of this study was to detect the IS900 region of Mycobacterium avium subsp. paratuberculosis (MAP) in bovine milk samples using real-time polymerase chain reaction (qPCR) and conventional PCR, and to study the agreement between these tests. A total of 121 bovine milk samples were collected from herds considered positive for MAP, from the State of Pernambuco, Brazil. MAP DNA was detected in 20 samples (16.5%) using conventional PCR and in 34 samples (28.1%) using qPCR. MAP DNA was detected in all of the 6 animal farms studied. Moderate agreement was found between qPCR and conventional PCR results, where the sensitivity and specificity of conventional PCR in relation to qPCR were 50% and 96.6%, respectively. Thus, the IS900 region of MAP was found in bovine milk samples from the State of Pernambuco. To the best of our knowledge, this is the first report of MAP DNA found in bovine milk in Northeast Brazil. We also demonstrated the qPCR technique is more sensitive than conventional PCR with respect to detection of MAP in milk samples. Copyright © 2016. Published by Elsevier Editora Ltda.

  3. Rapid detection and subtyping of human influenza A viruses and reassortants by pyrosequencing.

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    Yi-Mo Deng

    Full Text Available BACKGROUND: Given the continuing co-circulation of the 2009 H1N1 pandemic influenza A viruses with seasonal H3N2 viruses, rapid and reliable detection of newly emerging influenza reassortant viruses is important to enhance our influenza surveillance. METHODOLOGY/PRINCIPAL FINDINGS: A novel pyrosequencing assay was developed for the rapid identification and subtyping of potential human influenza A virus reassortants based on all eight gene segments of the virus. Except for HA and NA genes, one universal set of primers was used to amplify and subtype each of the six internal genes. With this method, all eight gene segments of 57 laboratory isolates and 17 original specimens of seasonal H1N1, H3N2 and 2009 H1N1 pandemic viruses were correctly matched with their corresponding subtypes. In addition, this method was shown to be capable of detecting reassortant viruses by correctly identifying the source of all 8 gene segments from three vaccine production reassortant viruses and three H1N2 viruses. CONCLUSIONS/SIGNIFICANCE: In summary, this pyrosequencing assay is a sensitive and specific procedure for screening large numbers of viruses for reassortment events amongst the commonly circulating human influenza A viruses, which is more rapid and cheaper than using conventional sequencing approaches.

  4. Detection and differentiation of Cryptosporidium by real-time polymerase chain reaction in stool samples from patients in Rio de Janeiro, Brazil

    Directory of Open Access Journals (Sweden)

    Roberta Flávia Ribeiro Rolando

    2012-06-01

    Full Text Available This study reports the first genetic characterisation of Cryptosporidium isolates in Brazil using real-time polymerase chain reaction (RT-PCR. A total of 1,197 faecal specimens from children and 10 specimens from human immunodeficiency virus-infected patients were collected between 1999-2010 and screened using microscopy. Forty-eight Cryptosporidium oocyst-positive isolates were identified and analysed using a generic TaqMan assay targeting the 18S rRNA to detect Cryptosporidium species and two other TaqMan assays to identify Cryptosporidium hominis and Cryptosporidium parvum. The 18S rRNA assay detected Cryptosporidium species in all 48 of the stool specimens. The C. parvum TaqMan assay correctly identified five/48 stool samples, while 37/48 stool specimens were correctly amplified in the C. hominis TaqMan assay. The results obtained in this study support previous findings showing that C. hominis infections are more prevalent than C. parvum infections in Brazil and they demonstrate that the TaqMan RT-PCR procedure is a simple, fast and valuable tool for the detection and differentiation of Cryptosporidium species.

  5. Prevalence of Human T-lymphotropic virus type 1 and 2 among blood donors in Manaus, Amazonas State, Brazil.

    Science.gov (United States)

    Morais, Márcia Poinho EncarnaçÃo de; Gato, Cynara Melo; Maciel, Lucinei Alves; Lalwani, Pritesh; Costa, Cristóvão Alves; Lalwani, Jaila Dias Borges

    2017-12-21

    Human T-lymphotropic virus type 1 and 2 (HTLV-1/2) is endemic in Brazil, but few studies have investigated the seroprevalence of HTLV and its subtypes among blood donors in the capital city Manaus, Amazonas State, Brazil. To estimate the seroprevalence of HTLV-1/2 and to identify circulating subtypes among blood donors in Manaus. Blood donors (2001-2003) were screened for HTLV-1/2 antibodies by ELISA. Positive results were confirmed and subtyped by Western blot assays. Prevalence rates were calculated and compared with demographic data. Among the 87,402 individuals screened, 116 (0.13%) were seropositive for HTLV-1/2. A second sample (76/116) was collected and retested by HTLV-1/2 ELISA, of which only 41/76 were positive. Western blot confirmed HTLV infection in 24/41 retested blood donors [HTLV-1 (n=16), HTLV-2 (n=5) and HTLV-untypable (n=3)]. HTLV-1 and HTLV-2 are prevalent among blood donors in Manaus. However, additional studies are needed to comprehend the epidemiology of HTLV-1/2 in Amazonas not only to understand the pathophysiology of the disease providing adequate medical assistance, but also to reduce or block virus transmission.

  6. Prevalence of Human T-lymphotropic virus type 1 and 2 among blood donors in Manaus, Amazonas State, Brazil

    Directory of Open Access Journals (Sweden)

    Márcia Poinho EncarnaçÃo de Morais

    2017-12-01

    Full Text Available ABSTRACT Introduction: Human T-lymphotropic virus type 1 and 2 (HTLV-1/2 is endemic in Brazil, but few studies have investigated the seroprevalence of HTLV and its subtypes among blood donors in the capital city Manaus, Amazonas State, Brazil. Aim: To estimate the seroprevalence of HTLV-1/2 and to identify circulating subtypes among blood donors in Manaus. Materials and Methods: Blood donors (2001-2003 were screened for HTLV-1/2 antibodies by ELISA. Positive results were confirmed and subtyped by Western blot assays. Prevalence rates were calculated and compared with demographic data. Results: Among the 87,402 individuals screened, 116 (0.13% were seropositive for HTLV-1/2. A second sample (76/116 was collected and retested by HTLV-1/2 ELISA, of which only 41/76 were positive. Western blot confirmed HTLV infection in 24/41 retested blood donors [HTLV-1 (n=16, HTLV-2 (n=5 and HTLV-untypable (n=3]. Discussion: HTLV-1 and HTLV-2 are prevalent among blood donors in Manaus. However, additional studies are needed to comprehend the epidemiology of HTLV-1/2 in Amazonas not only to understand the pathophysiology of the disease providing adequate medical assistance, but also to reduce or block virus transmission.

  7. Zika virus infection, associated microcephaly, and low yellow fever vaccination coverage in Brazil: is there any causal link?

    Science.gov (United States)

    De Góes Cavalcanti, Luciano Pamplona; Tauil, Pedro Luiz; Alencar, Carlos Henrique; Oliveira, Wanderson; Teixeira, Mauro Martins; Heukelbach, Jorg

    2016-06-30

    Since the end of 2014, Zika virus (ZIKV) infection has been rapidly spreading in Brazil. To analyze the possible association of yellow fever vaccine with a protective effect against ZIKV-related microcephaly, the following spatial analyses were performed, using Brazilian municipalities as units: i) yellow fever vaccination coverage in Brazilian municipalities in individuals aged 15-49; ii) reported cases of microcephaly by municipality; and iii) confirmed cases of microcephaly related to ZIKV, by municipality. SaTScan software was used to identify clusters of municipalities for high risk of microcephaly. There were seven significant high risk clusters of confirmed microcephaly cases, with four of them located in the Northeast where yellow fever vaccination rates were the lowest. The clusters harbored only 2.9% of the total population of Brazil, but 15.2% of confirmed cases of microcephaly. We hypothesize that pregnant women in regions with high yellow fever vaccination coverage may pose their offspring to lower risk for development of microcephaly. There is an urgent need for systematic studies to confirm the possible link between low yellow fever vaccination coverage, Zika virus infection and microcephaly.

  8. Molecular surveillance of dengue in Minas Gerais provides insights on dengue virus 1 and 4 circulation in Brazil.

    Science.gov (United States)

    Dutra, Karina Rocha; Drumond, Betânia Paiva; de Rezende, Izabela Maurício; Nogueira, Maurício Lacerda; de Oliveira Lopes, Débora; Calzavara Silva, Carlos Eduardo; Siqueira Ferreira, Jaqueline Maria; Dos Santos, Luciana Lara

    2017-06-01

    Dengue, caused by any of the four types of Dengue virus (DENV) is the most important arbovirus in the world. In this study we performed a molecular surveillance of dengue during the greatest dengue outbreak that took place in Divinópolis, Minas Gerais state, Southeast Brazil, in 2013. Samples from 100 patients with clinical symptoms of dengue were studied and 26 were positive. The capsid/premembrane (CprM) and envelope gene sequences of some samples were amplified and sequenced. Molecular analyses demonstrated that two DENV-1 lineages, belonging to genotype V were introduced and co-circulated in Divinópolis. When compared to each other, those lineages presented high genetic diversity and showed unique amino acids substitutions in the envelope protein, including in domains I, II, and III. DENV-4 strains from Divinópolis clustered within genotype IIb and the most recent common ancestor was probably introduced into the city three years before the 2013 epidemic. Here we demonstrated for the first time the circulation of DENV-4 and the co-circulation of two DENV-1 lineages in Midwest region of Minas Gerais, Brazil. Moreover our analysis indicated the introduction of five DENV-1 lineages, genotype V into Brazil, in different times. J. Med. Virol. 89:966-973, 2017. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  9. Multiplex Real Time PCR For Detection of Wheat Streak Mosaic Virus and Triticum Mosaic Virus

    Science.gov (United States)

    Wheat streak mosaic virus (WSMV) and Triticum mosaic virus (TRIMV) are widespread throughout the southwestern Great Plains states. Using conventional diagnostics such as Enzyme-Linked Immunosorbent Assays (ELISA), these two viruses are commonly found together in infected wheat samples. Methods for m...

  10. Perinatal hepatitis B virus detection by hepatitis B virus-DNA analysis.

    OpenAIRE

    de Virgiliis, S; Frau, F; G. Sanna; Turco, M P; Figus, A L; Cornacchia, G; Cao, A.

    1985-01-01

    Maternal transmission of hepatitis B virus infection in relation to the hepatitis B e antigen/antibody system and serum hepatitis B virus-DNA were evaluated. Results indicate that hepatitis B virus-DNA analysis can identify hepatitis B serum antigen positive mothers who may transmit infection to their offspring.

  11. Rhinovirus-C detection in children presenting with acute respiratory infection to hospital in Brazil.

    Science.gov (United States)

    Fawkner-Corbett, David W; Khoo, Siew Kim; Duarte, Carminha M; Bezerra, Patricia G M; Bochkov, Yury A; Gern, James E; Le Souef, Peter N; McNamara, Paul S

    2016-01-01

    Human rhinovirus (RV) is a common cause of acute respiratory infection (ARI) in children. We aimed to characterize the clinical and demographic features associated with different RV species detected in children attending hospital with ARI, from low-income families in North-east Brazil. Nasopharyngeal aspirates were collected from 630 children infected with RV-A and RV-C. However, in children with asthma/EVW, RV-C was detected relatively more frequently than RV-A (23% vs. 5%; P = 0.04). Our findings highlight RV as a potentially important pathogen in this setting. Generally, clinical and demographic features were similar in children in whom RV-A and C species were detected. However, RV-C was more frequently found in children with asthma/EVW than RV-A. © 2015 Wiley Periodicals, Inc.

  12. Comprehensive detection of viruses in pediatric patients with acute liver failure using next-generation sequencing.

    Science.gov (United States)

    Suzuki, Takako; Kawada, Jun-Ichi; Okuno, Yusuke; Hayano, Satoshi; Horiba, Kazuhiro; Torii, Yuka; Takahashi, Yoshiyuki; Umetsu, Syuichiro; Sogo, Tsuyoshi; Inui, Ayano; Ito, Yoshinori

    2017-11-01

    Pediatric acute liver failure (PALF) is a rare and severe syndrome that frequently requires liver transplantation. Viruses are one of the most frequent causes of this disease, however, pathogenic viruses are not determined in many patients. Recently next-generation sequencing (NGS) has been applied to comprehensively detect pathogens of infectious diseases of unknown etiology. To evaluate an NGS-based approach for detecting pathogenic viruses in patients with PALF or acute hepatitis of unknown etiology. To detect virus-derived DNA and RNA sequences existing in sera/plasma from patients, both DNA and RNA sequencing were performed. First, we validated the ability of NGS to detect viral pathogens in clinical serum/plasma samples, and compared different commercial RNA library preparation methods Then, serum/plasma of fourteen patients with PALF or acute hepatitis of unknown etiology were evaluated using NGS. Among three RNA library preparation methods, Ovation RNA-Seq System V2 had the highest sensitivity to detect RNA viral sequences. Among fourteen patients, sequence reads of torque teno virus, adeno-associated virus, and stealth virus were found in the sera of one patient each, however, the pathophysiological role of these three viruses was not clarified. Significant virus reads were not detected in the remaining 11 patients. This finding might be due to low virus titer in blood at the time of referral or a non-infectious cause might be more frequent. These results suggest an NGS-based approach has potential to detect viral pathogens in clinical samples and would contribute to clarification of the etiology of PALF. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Detection of Onchocerca volvulus (Nematoda: Onchocercidae infection in vectors from Amazonian Brazil following mass Mectizan™ distribution

    Directory of Open Access Journals (Sweden)

    Verônica Marchon-Silva

    2007-03-01

    Full Text Available Detection of Onchocerca volvulus in Simulium populations is of primary importance in the assessment of the effectiveness of onchocerciasis control programs. In Brazil, the main focus of onchocerciasis is in the Amazon region, in a Yanomami reserve. The main onchocerciasis control strategy in Brazil is the semi-annually mass distribution of the microfilaricide ivermectin. In accordance with the control strategy for the disease, polymerase chain reaction (PCR was applied in pools of simuliids from the area to detect the helminth infection in the vectors, as recommended by the Onchocerciasis Elimination Program for the Americas and the World Health Organization. Systematic sampling was performed monthly from September 1998 to October 1999, and a total of 4942 blackflies were collected from two sites (2576 from Balawaú and 2366 from Toototobi. The molecular methodology was found to be highly sensitive and specific for the detection of infected and/or infective blackflies in pools of 50 blackflies. The results from the material collected under field conditions showed that after the sixth cycle of distribution of ivermectin, the prevalence of infected blackflies with O. volvulus had decreased from 8.6 to 0.3% in Balawaú and from 4 to 0.1% in Toototobi.

  14. First detection of adenovirus in the vampire bat (Desmodus rotundus) in Brazil.

    Science.gov (United States)

    Lima, Francisco Esmaile de Sales; Cibulski, Samuel Paulo; Elesbao, Felipe; Carnieli Junior, Pedro; Batista, Helena Beatriz de Carvalho Ruthner; Roehe, Paulo Michel; Franco, Ana Cláudia

    2013-10-01

    This paper describes the first detection of adenovirus in a Brazilian Desmodus rotundus bat, the common vampire bat. As part of a continuous rabies surveillance program, three bat specimens were captured in Southern Brazil. Total DNA was extracted from pooled organs and submitted to a nested PCR designed to amplify a 280 bp long portion of the DNA polymerase gene of adenoviruses. One positive sample was subjected to nucleotide sequencing, confirming that this DNA fragment belongs to a member of the genus Mastadenovirus. This sequence is approximately 25 % divergent at the nucleotide level from equine adenovirus 1 and two other recently characterized bat adenoviruses.

  15. Detection of antibodies against Leishmania infantum in cats (Felis catus) from the State of Pernambuco, Brazil.

    Science.gov (United States)

    Silva, Rita de Cássia Nascimento; Ramos, Rafael Antonio Nascimento; Pimentel, Danillo de Souza; Oliveira, Gênova Maria de Azevedo; Carvalho, Gílcia Aparecida de; Santana, Marília de Andrade; Faustino, Maria Aparecida da Glória; Alves, Leucio Câmara

    2014-01-01

    Little information is available concerning infection by Leishmania infantum in cats. Therefore, the aim of this study was to perform a serological study in domestic cats. Serum samples (n=153) obtained from animals living in the Cities of Recife and Petrolina, State of Pernambuco, Brazil, were tested by ELISA/S7® (Biogene). Anti-L. infantum antibodies were detected in 3.9% (6/153) of the cats. All seroreagent animals were from Petrolina. These results serve as an important alert, and future studies are needed to better understand the possible role of cats in the epidemiology of visceral leishmaniasis (VL) in this area.

  16. Detection of psychiatric morbidity in the primary medical care setting in Brazil

    OpenAIRE

    Mari,Jair de Jesus; Iacoponi,Eduardo; Williams,Paul; Simões,Oziris; Silva,João Batista Teodoro

    1987-01-01

    The aims of this study were a) to assess the ability of primary care doctors to make accurate ratings of psychiatric disturbance and b) to evaluate the use of a case-finding questionnaire in the detection of psychiatric morbidity. The estudy took place in three primary care clinics in the city of São Paulo, Brazil, during a six-month survey. A time sample of consecutive adult attenders were asked to complete a case-finding questionnaire for psychiatric disorders (the Self Report Questionnaire...

  17. Plasmonic Nanomaterial-Based Optical Biosensing Platforms for Virus Detection.

    Science.gov (United States)

    Lee, Jaewook; Takemura, Kenshin; Park, Enoch Y

    2017-10-13

    Plasmonic nanomaterials (P-NM) are receiving attention due to their excellent properties, which include surface-enhanced Raman scattering (SERS), localized surface plasmon resonance (LSPR) effects, plasmonic resonance energy transfer (PRET), and magneto optical (MO) effects. To obtain such plasmonic properties, many nanomaterials have been developed, including metal nanoparticles (MNP), bimetallic nanoparticles (bMNP), MNP-decorated carbon nanotubes, (MNP-CNT), and MNP-modified graphene (MNP-GRP). These P-NMs may eventually be applied to optical biosensing systems due to their unique properties. Here, probe biomolecules, such as antibodies (Ab), probe DNA, and probe aptamers, were modified on the surface of plasmonic materials by chemical conjugation and thiol chemistry. The optical property change in the plasmonic nanomaterials was monitored based on the interaction between the probe biomolecules and target virus. After bioconjugation, several optical properties, including fluorescence, plasmonic absorbance, and diffraction angle, were changed to detect the target biomolecules. This review describes several P-NMs as potential candidates of optical sensing platforms and introduces various applications in the optical biosensing field.

  18. Plasmonic Nanomaterial-Based Optical Biosensing Platforms for Virus Detection

    Directory of Open Access Journals (Sweden)

    Jaewook Lee

    2017-10-01

    Full Text Available Plasmonic nanomaterials (P-NM are receiving attention due to their excellent properties, which include surface-enhanced Raman scattering (SERS, localized surface plasmon resonance (LSPR effects, plasmonic resonance energy transfer (PRET, and magneto optical (MO effects. To obtain such plasmonic properties, many nanomaterials have been developed, including metal nanoparticles (MNP, bimetallic nanoparticles (bMNP, MNP-decorated carbon nanotubes, (MNP-CNT, and MNP-modified graphene (MNP-GRP. These P-NMs may eventually be applied to optical biosensing systems due to their unique properties. Here, probe biomolecules, such as antibodies (Ab, probe DNA, and probe aptamers, were modified on the surface of plasmonic materials by chemical conjugation and thiol chemistry. The optical property change in the plasmonic nanomaterials was monitored based on the interaction between the probe biomolecules and target virus. After bioconjugation, several optical properties, including fluorescence, plasmonic absorbance, and diffraction angle, were changed to detect the target biomolecules. This review describes several P-NMs as potential candidates of optical sensing platforms and introduces various applications in the optical biosensing field.

  19. Unbiased approach for virus detection in skin lesions.

    Directory of Open Access Journals (Sweden)

    Davit Bzhalava

    Full Text Available To assess presence of virus DNA in skin lesions, swab samples from 82 squamous cell carcinomas of the skin (SCCs, 60 actinic keratoses (AKs, paraffin-embedded biopsies from 28 SCCs and 72 kerathoacanthomas (KAs and fresh-frozen biopsies from 92 KAs, 85 SCCs and 92 AKs were analyzed by high throughput sequencing (HTS using 454 or Ion Torrent technology. We found total of 4,284 viral reads, out of which 4,168 were Human Papillomavirus (HPV-related, belonging to 15 known (HPV8, HPV12, HPV20, HPV36, HPV38, HPV45, HPV57, HPV59, HPV104, HPV105, HPV107, HPV109, HPV124, HPV138, HPV147, four previously described putative (HPV 915 F 06 007 FD1, FA73, FA101, SE42 and two putatively new HPV types (SE46, SE47. SE42 was cloned, sequenced, designated as HPV155 and found to have 76% similarity to the most closely related known HPV type. In conclusion, an unbiased approach for viral DNA detection in skin tumors has found that, although some new putative HPVs were found, known HPV types constituted most of the viral DNA.

  20. Detection of late intermediates in virus capsid assembly by charge detection mass spectrometry.

    Science.gov (United States)

    Pierson, Elizabeth E; Keifer, David Z; Selzer, Lisa; Lee, Lye Siang; Contino, Nathan C; Wang, Joseph C-Y; Zlotnick, Adam; Jarrold, Martin F

    2014-03-05

    The assembly of hundreds of identical proteins into an icosahedral virus capsid is a remarkable feat of molecular engineering. How this occurs is poorly understood. Key intermediates have been anticipated at the end of the assembly reaction, but it has not been possible to detect them. In this work we have used charge detection mass spectrometry to identify trapped intermediates from late in the assembly of the hepatitis B virus T = 4 capsid, a complex of 120 protein dimers. Prominent intermediates are found with 104/105, 110/111, and 117/118 dimers. Cryo-EM observations indicate the intermediates are incomplete capsids and, hence, on the assembly pathway. On the basis of their stability and kinetic accessibility we have proposed plausible structures. The prominent trapped intermediate with 104 dimers is attributed to an icosahedron missing two neighboring facets, the 111-dimer species is assigned to an icosahedron missing a single facet, and the intermediate with 117 dimers is assigned to a capsid missing a ring of three dimers in the center of a facet.

  1. Functionalized magnetic microparticle-based colorimetric platform for influenza A virus detection

    Science.gov (United States)

    Chen, Chaohui; Zou, Zhong; Chen, Lu; Ji, Xinghu; He, Zhike

    2016-10-01

    A colorimetric platform for influenza A virus detection was developed by using the high efficiency of enzymatic catalysis and the reduction of gold ions with hydrogen peroxide. Aptamer-functionalized magnetic microparticles were synthesized to capture the influenza A virus. This was followed by the binding of ConA-GOx-AuNPs to the H3N2 virus through the ConA-glycan interaction. The sandwich complex was subsequently dispersed in glucose solution to trigger an enzymatic reaction to produce hydrogen peroxide, which controlled the growth of gold nanoparticles and produced colored solutions. The determination of H3N2 concentration was realized by comparing the two differently colored gold nanoparticles. This method could detect the target virus as low as 11.16 μg ml-1. Furthermore, it opens new opportunities for sensitive and colorimetric detection of viruses and proteins.

  2. An improved electrochemiluminescence polymerase chain reaction method for highly sensitive detection of plant viruses

    Energy Technology Data Exchange (ETDEWEB)

    Tang Yabing [MOE Key Laboratory of Laser Life Science and Institute of Laser Life Science, South China Normal University, Guangzhou 510631 (China); Xing Da [MOE Key Laboratory of Laser Life Science and Institute of Laser Life Science, South China Normal University, Guangzhou 510631 (China)]. E-mail: xingda@scnu.edu.cn; Zhu Debin [MOE Key Laboratory of Laser Life Science and Institute of Laser Life Science, South China Normal University, Guangzhou 510631 (China); Liu Jinfeng [MOE Key Laboratory of Laser Life Science and Institute of Laser Life Science, South China Normal University, Guangzhou 510631 (China)

    2007-01-23

    Recently, we have reported an electrochemiluminescence polymerase chain reaction (ECL-PCR) method for detection of genetically modified organisms. The ECL-PCR method was further improved in the current study by introducing a multi-purpose nucleic acid sequence that was specific to the tris(bipyridine) ruthenium (TBR) labeled probe, into the 5' terminal of the primers. The method was applied to detect plant viruses. Conserved sequence of the plant viruses was amplified by PCR. The product was hybridized with a biotin labeled probe and a TBR labeled probe. The hybridization product was separated by streptavidin-coated magnetic beads, and detected by measuring the ECL signals of the TBR labeled. Under the optimized conditions, the experiment results show that the detection limit is 50 fmol of PCR products, and the signal-to-noise ratio is in excess of 14.6. The method was used to detect banana streak virus, banana bunchy top virus, and papaya leaf curl virus. The experiment results show that this method could reliably identity viruses infected plant samples. The improved ECL-PCR approach has higher sensitivity and lower cost than previous approach. It can effectively detect the plant viruses with simplicity, stability, and high sensitivity.

  3. Respiratory viruses are continuously detected in children with chronic tonsillitis throughout the year.

    Science.gov (United States)

    Proença-Módena, José Luiz; Buzatto, Guilherme P; Paula, Flávia E; Saturno, Tamara H; Delcaro, Luana S; Prates, Mirela C; Tamashiro, Edwin; Valera, Fabiana C P; Arruda, Eurico; Anselmo-Lima, Wilma T

    2014-10-01

    To evaluate the oscillations on the viral detection in adenotonsillar tissues from patients with chronic adenotonsillar diseases as an indicia of the presence of persistent viral infections or acute subclinical infections. Cross-sectional prospective study. Tertiary hospital. The fluctuations of respiratory virus detection were compared to the major climatic variables during a two-year period using adenoids and palatine tonsils from 172 children with adenotonsillar hypertrophy and clinical evidence of obstructive sleep apnoea syndrome or recurrent adenotonsillitis, without symptoms of acute respiratory infection (ARI), by TaqMan real-time PCR. The rate of detection of at least one respiratory virus in adenotonsillar tissue was 87%. The most frequently detected viruses were human adenovirus in 52.8%, human enterovirus in 47.2%, human rhinovirus in 33.8%, human bocavirus in 31.1%, human metapneumovirus in 18.3% and human respiratory syncytial virus in 17.2%. Although increased detection of human enterovirus occurred in summer/autumn months, and there were summer nadirs of human respiratory syncytial virus in both years of the study, there was no obvious viral seasonality in contrast to reports with ARI patients in many regions of the world. Respiratory viruses are continuously highly detected during whole year, and without any clinical symptomatology, indicating that viral genome of some virus can persist in lymphoepithelial tissues of the upper respiratory tract. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  4. Investigation of Influenza A, West Nile and Newcastle Disease Viruses in Birds from the Pantanal Wetlands of Mato Grosso, Brazil

    Directory of Open Access Journals (Sweden)

    LB Pinto

    Full Text Available ABSTRACT The Pantanal is the world's largest wetland biome with a seasonal flood pulse that attracts a great diversity of birds, many of which are migratory. Birds can be natural reservoirs Influenza A, West Nile and Newcastle Disease viruses. However, the occurrence of carriers for these viruses in the Pantanal was not verified yet. The present study evaluated the occurrence of natural infection by Influenza A, WN and ND virus of birds in the municipality of Poconé, a subregion of the Pantanal in the state of Mato Grosso, Brazil. A total of 76 birds belonging to 11 orders and 20 families were captured using mist nets. The most representative order was Passeriformes, followed by the other nine orders, which included Columbiformes, Psittaciformes, Charadriiformes and Anseriformes. The most representative family was Thamnophilidae, with 16 individuals (21.0%, followed by the family Tyrannidae with 10 individuals (7.6% and the family Furnariidae, with eight individuals (10.5%. The bird species were identified, and cloacal and tracheal swab samples were collected. The samples were subjected to RNA extraction and tested for the presence of the three agents by real-time polymerase chain reaction (qRT-PCR. All the sampled birds were considered healthy, had no clinical sign of infection, and were tested negative for the three viruses. Based on our findings, we can conclude that Influenza, West Nile and Newcastle Disease viruses were absent from the samples in this region of the Pantanal wetlands during the period of this study.

  5. A Novel Astrovirus-Like RNA Virus Detected in Human Stool

    NARCIS (Netherlands)

    Oude Munnink, Bas B.; Cotten, Matthew; Canuti, Marta; Deijs, Martin; Jebbink, Maarten F.; van Hemert, Formijn J.; Phan, My V. T.; Bakker, Margreet; Jazaeri Farsani, Seyed Mohammad; Kellam, Paul; van der Hoek, Lia

    2016-01-01

    Several novel clades of astroviruses have recently been identified in human faecal samples. Here, we describe a novel astrovirus-like RNA virus detected in human stools, which we have tentatively named bastrovirus. The genome of this novel virus consists of 6,300 nucleotides organized in three open

  6. Detecting the Presence of Nora Virus in "Drosophila" Utilizing Single Fly RT-PCR

    Science.gov (United States)

    Munn, Bethany; Ericson, Brad; Carlson, Darby J.; Carlson, Kimberly A.

    2015-01-01

    A single fly RT-PCR protocol has recently been developed to detect the presence of the persistent, horizontally transmitted Nora virus in "Drosophila." Wild-caught flies from Ohio were tested for the presence of the virus, with nearly one-fifth testing positive. The investigation presented can serve as an ideal project for biology…

  7. Screen-Printed All-Polymer Aptasensor for Impedance Based Detection of Influenza A Virus

    DEFF Research Database (Denmark)

    Kirkegaard, Julie; Rozlosnik, Noemi

    2017-01-01

    are made by CO2 laser cutting of Poly(methyl methacrylate) (PMMA) sheets. Influenza A virus specific aptamers are immobilized onto the electrodes by UV cross-linking. Impedance based measurements at a single frequency, measured over time, are used to detect the virus in a buffer solution....

  8. A sensitive cell-based assay for the detection of residual infectious West Nile virus

    NARCIS (Netherlands)

    Koldijk, M. H.; Bogaards, J. A.; Kostense, S.; de Vocht, M.; Gijsbers, L.; ter Haak, M.; Ophorst, C.; Brakenhoff, J. P. J.; Weverling, G. J.; Guichoux, J. Y.; Uytdehaag, F.; Lewis, J.; Goudsmit, J.; Marzio, G.

    2007-01-01

    Ensuring complete viral inactivation is critical for the safety of vaccines based on an inactivated virus. Detection of residual infectious virus is dependent on sensitivity of the assay, sample volume analyzed and the absence of interference with viral infection. Here we describe the development

  9. 35 original article detection of influenza a virus in pigs in lagos, nigeria

    African Journals Online (AJOL)

    User

    (16). The high number of positive results to influenza A virus 31(26.7%) detected in this study is characterized by its continuous antigenic drift and shift, making it more genetically diverse with high prevalence in the subject (8,4). This is further supported by the work of. (17) which showed that influenza A virus mutates at a.

  10. Comparative detection of a large population of grapevine viruses by TaqMan® RT-qPCR and ELISA.

    Science.gov (United States)

    Bruisson, Sébastien; Lebel, Sylvain; Walter, Bernard; Prevotat, Laurent; Seddas, Sam; Schellenbaum, Paul

    2017-02-01

    Grapevine (Vitis spp.) can be infected by numerous viruses that are often widespread and of great economic importance. Reliable detection methods are necessary for sanitary selection which is the only way to partly control grapevine virus diseases. Biological indexing and ELISA are currently the standard methods for screening propagation material, and PCR-methods are becoming increasingly popular. Due to the diversity of virus isolates, it is essential to verify that the tests allow the detection of the largest possible virus populations. We developed three quadruplex TaqMan® RT-qPCR assays for detecting nine different viruses that cause considerable damage in many vineyards world-wide. Each assay is designed to detect three viruses and the grapevine Actin as an internal control. A large population of grapevines from diverse cultivars and geographic location was tested for the presence of nine viruses: Arabis mosaic virus (ArMV), Grapevine fleck virus (GFkV), Grapevine fanleaf virus (GFLV), Grapevine leafroll-associated viruses (GLRaV-1, -2, -3), Grapevine rupestris stem pitting-associated virus (GRSPaV), Grapevine virus A (GVA), and Grapevine virus B (GVB). In general, identical results were obtained with multiplex TaqMan® RT-qPCR and ELISA although, in some cases, viruses could be detected by only one of the two techniques. Copyright © 2016 Elsevier B.V. All rights reserved.

  11. Duplex PCR assay for the detection of avian adeno virus and chicken anemia virus prevalent in Pakistan

    Directory of Open Access Journals (Sweden)

    Iqbal Aqib

    2011-09-01

    Full Text Available Abstract Avian Adeno viruses and Chicken Anemia Viruses cause serious economic losses to the poultry industry of Pakistan each year. Timely and efficient diagnosis of the viruses is needed in order to practice prevention and control strategies. In the first part of this study, we investigated broilers, breeder and Layer stocks for morbidity and mortality rates due to AAV and CAV infections and any co-infections by examining signs and symptoms typical of their infestation or post mortem examination. In the second part of the study, we developed a duplex PCR assay for the detection of AAV and CAV which is capable to simultaneously detect both the viral types prevalent in Pakistan with high sensitivity and 100% specificity.

  12. The detection of Vaccinia virus confirms the high circulation of Orthopoxvirus in buffaloes living in geographical isolation, Marajó Island, Brazilian Amazon.

    Science.gov (United States)

    Franco-Luiz, Ana Paula Moreira; Fagundes Pereira, Alexandre; de Oliveira, Cairo Henrique Sousa; Barbosa, José Diomedes; Oliveira, Danilo Bretas; Bonjardim, Cláudio Antônio; Ferreira, Paulo César Peregrino; de Souza Trindade, Giliane; Abrahão, Jônatas Santos; Kroon, Erna Geessien

    2016-06-01

    In Brazil, serologic evidence of Orthopoxvirus (OPV) circulation showed positivity around 20% in cattle, humans, monkeys and rodents. Although OPV seropositivity has been described in buffalo herds in southeastern Brazil, no Vaccinia virus (VACV) (member of genus OPV) outbreaks in buffalo herds have been described in this country. This study aimed to investigate the detection of anti-OPV antibodies and to study the OPV genome in Brazilian buffalo herds. Our results demonstrated a high OPV seropositivity in buffalo herds on Marajó Island and molecular data confirmed the circulation of VACV. The geographical isolation conditionmight be a sine qua non condition to explain our results. Copyright © 2016. Published by Elsevier Ltd.

  13. External Quality Assessment of Molecular Detection of Ebola Virus in China

    Science.gov (United States)

    Wang, Guojing; Sun, Yu; Zhang, Kuo; Jia, Tingting; Hao, Mingju; Zhang, Dong; Chang, Le; Zhang, Lei; Zhang, Rui; Lin, Guigao; Peng, Rongxue; Li, Jinming

    2015-01-01

    In 2014, Ebola hemorrhagic fever broke out in West Africa. As contact between China and West Africa is frequent, the possibility that Ebola virus would enter China was high. Thus, an external assessment of the quality of Ebola virus detection was organized by the National Center for Clinical Laboratories in China. Virus-like particles encapsulating known sequences of epidemic strains of Ebola virus from 2014 were prepared as positive quality controls. The sample panel, which was composed of seven positive and three negative samples, was dispatched to 19 laboratories participating in this assessment of Ebola virus detection. Accurate detection was reported at 14 of the 19 participating laboratories, with a sensitivity of 91.43% and a specificity of 100%. Four participants (21.05%) reported false-negative results and were classified as “acceptable.” One participant (5.26%) did not detect any positive samples and was thus classified as “improvable.” Based on the results returned, the ability to detect weakly positive Ebola specimens should be improved. Furthermore, commercial assays and the standard primers offered by the Chinese Centers for Disease Control and Prevention were found to be most accurate and dependable for Ebola detection. A two-target detection approach is recommended for Ebola screening; this approach could reduce the probability of false-negative results. Additionally, standardization of operations and punctual adjustment of instruments are necessary for the control and prevention of Ebola virus. PMID:26177537

  14. New target tissue for food-borne virus detection in oysters.

    Science.gov (United States)

    Wang, D; Wu, Q; Yao, L; Wei, M; Kou, X; Zhang, J

    2008-11-01

    To evaluate the different tissues of naturally contaminated oyster for food-borne virus detection. The different tissues of 136 field oyster samples were analysed for norovirus (NV), hepatitis A virus (HAV) and rotavirus (RV) by reverse transcription (RT)-PCR and were confirmed by sequencing. These viruses were detected in 20 samples (14.71%), showing positivity for NV (1.47%), HAV (5.15%) and RV (8.82%). Furthermore, among different tissues, the highest positive rate of the food-borne viruses was found in the gills (14.71%), followed by the stomach (13.97%) and the digestive diverticula (13.24%). The food-borne viruses were detected in the gills, stomach, digestive diverticula and the cilia of the mantle. In addition, the results showed that the gills are one of the appropriate tissues for viral detection in oysters by nucleic acid assay. This is the first paper to report on the presence of food-borne viruses in the gills and the cilia of the mantle of naturally contaminated oysters. The research team hopes that the results of the study will be of help in sampling the appropriate tissues for the detection of food-borne viruses in commercial oysters.

  15. Detection and characterization of fibropapilloma associated herpesvirus of marine turtles in Rio Grande do Sul, Brazil

    Directory of Open Access Journals (Sweden)

    Carla R. Rodenbusch

    2012-11-01

    Full Text Available Fibropapillomatosis (FP is a benign tumoral disease that affects sea turtles, hampering movement, sight and feeding, ultimately leading to death. In Brazil, the disease was described for the first time in 1986. Research suggests the involvement of a herpesvirus in association with environmental and genetic factors as causal agents of FP. The objective of the present study was to detect and characterize this herpesvirus in sea turtles living in the coast of state Rio Grande do Sul (RS, Brazil. From October 2008 to July 2010, 14 turtles were observed between the beaches of Torres and Tavares, of which 11 were green turtles (Chelonia mydas and 3 were loggerhead turtles (Caretta caretta. All turtles were young and mean curved carapace length was 37.71±7.82cm, and varied from 31 to 55cm. Only one green turtle presented a 1cm, papillary, pigmented fibropapilloma. Skin and fibropapilloma samples were analyzed by conventional and real time PCR assays to detect and quantify herpesvirus. All skin samples were negative, though the fibropapilloma specimen was positive in both tests. Viral load was 9,917.04 copies of viral genome per milligram of tissue. The DNA fragment amplified from the fibropapilloma sample was sequenced and allocated in the Atlantic phylogeographic group. This study reports the first molecular characterization of herpesvirus associated with fibropapilloma in turtles from the coast of RS.

  16. [Development of A synthetic positive control for the nucleic acid detection of mumps virus].

    Science.gov (United States)

    Cui, Ai-li; Jin, Li; Xu, Wen-bo

    2013-09-01

    To rapidly identify the cross-contamination problems caused by the positive control in the process of mumps virus nucleic acid detection, a new mumps virus RNA positive control was developed in this study. Using the same primers and reaction conditions, the cross-contamination problems caused by the positive control could be readily identified by comparing the fragments lengths of the PCR products between the positive control and the samples. This new RNA positive control of mumps virus can be widely used in the diagnosis and genotyping of mumps virus as a better laboratory quality control.

  17. Round-robin comparison of methods for the detection of human enteric viruses in lettuce

    DEFF Research Database (Denmark)

    Le Guyader, Francoise S.; Schultz, Anna Charlotte; Haugarreau, Larissa

    2004-01-01

    Five methods that detect human enteric virus contamination in lettuce were compared. To mimic multiple contaminations as observed after sewage contamination, artificial contamination was with human calicivirus and poliovirus and animal calicivirus strains at different concentrations. Nucleic acid...

  18. Limited interlaboratory comparison of Schmallenberg virus antibody detection in serum samples.

    NARCIS (Netherlands)

    Poel, van der W.H.M.; Cay, B.; Zientara, S.; Steinbach, F.; Valarcher, J.F.; Botner, A.; Mars, M.H.; Hakze-van der Honing, van der R.W.; Schirrmeier, H.; Beer, M.

    2014-01-01

    Eight veterinary institutes in seven different countries in Europe participated in a limited interlaboratory comparison trial to evaluate laboratory performances of Schmallenberg virus (SBV) antibody detection in serum. Seven different sheep sera and three different cattle sera were circulated, and

  19. Notes from the Field: Evidence of Zika Virus Infection in Brain and Placental Tissues from Two Congenitally Infected Newborns and Two Fetal Losses--Brazil, 2015.

    Science.gov (United States)

    Martines, Roosecelis Brasil; Bhatnagar, Julu; Keating, M Kelly; Silva-Flannery, Luciana; Muehlenbachs, Atis; Gary, Joy; Goldsmith, Cynthia; Hale, Gillian; Ritter, Jana; Rollin, Dominique; Shieh, Wun-Ju; Luz, Kleber G; Ramos, Ana Maria de Oliveira; Davi, Helaine Pompeia Freire; Kleber de Oliveria, Wanderson; Lanciotti, Robert; Lambert, Amy; Zaki, Sherif

    2016-02-19

    Zika virus is a mosquito-borne flavivirus that is related to dengue virus and transmitted primarily by Aedes aegypti mosquitoes, with humans acting as the principal amplifying host during outbreaks. Zika virus was first reported in Brazil in May 2015 (1). By February 9, 2016, local transmission of infection had been reported in 26 countries or territories in the Americas.* Infection is usually asymptomatic, and, when symptoms are present, typically results in mild and self-limited illness with symptoms including fever, rash, arthralgia, and conjunctivitis. However, a surge in the number of children born with microcephaly was noted in regions of Brazil with a high prevalence of suspected Zika virus disease cases. More than 4,700 suspected cases of microcephaly were reported from mid-2015 through January 2016, although additional investigations might eventually result in a revised lower number (2). In response, the Brazil Ministry of Health established a task force to further investigate possible connections between the virus and brain anomalies in infants (3).

  20. An outbreak of post-vaccinal rabies (rage de laboratoire) in Fortaleza, Brazil, in 1960. Residual fixed virus as the etiological agent.

    Science.gov (United States)

    Pará, M

    1965-01-01

    The repeated isolation of fixed rabies virus from the CNS tissues of victims of an acute and lethal outbreak of encephalomyelitis in Fortaleza, Brazil, in November 1960, following vaccination with a locally produced killed-virus anti-rabies vaccine of the Fermi type is considered as definitive evidence of the rabic etiology (vaccinal fixed-virus rabies, rage de laboratoire) of this outbreak. Eighteen persons were affected, all of whom died.The clinical picture of paralytic rabies was recognizable in all of these 18 patients. The well-marked characteristics of an acute infection permit the easy differentiation of the paralysis caused by fixed rabies virus from post-vaccinal accidents that occur as allergic reactions.The incriminated anti-rabies vaccine was found to contain fixed live rabies virus at a titre of 10(-3.0). After one year of storage under refrigeration, the vaccine still contained fixed rabies virus, at a titre of 0,2x10(-1.0).Subsequent laboratory studies tend to indicate that the curve of inactivation of fixed virus by phenol does not follow a linear function but rather resembles the curve of inactivation of poliomyelitis virus by heat and formol according to the Salk technique. It is suggested that the antigenicity of the so-called "killed-virus" anti-rabies vaccines is actually due to the presence in them of residual amounts of live virus.

  1. Serological evidence of widespread circulation of West Nile virus and other flaviviruses in equines of the Pantanal, Brazil.

    Directory of Open Access Journals (Sweden)

    Alex Pauvolid-Corrêa

    2014-02-01

    Full Text Available A recent study reported neutralizing antibodies to West Nile virus (WNV in horses from four ranches of southern Pantanal. To extend that study, a serosurvey for WNV and 11 Brazilian flaviviruses was conducted with 760 equines, 238 sheep and 61 caimans from 17 local cattle ranches. Among the tested equines, 32 were collected from a ranch where a neurologic disorder outbreak had been recently reported. The sera were initially screened by using a blocking ELISA and then titrated by 90% plaque-reduction neutralization test (PRNT90 for 12 flaviviruses. Employing the criterion of 4-fold greater titer, 78 (10.3% equines were seropositive for Ilheus virus, 59 (7.8% for Saint Louis encephalitis virus, 24 (3.2% for WNV, two (0.3% for Cacipacore virus and one (0.1% for Rocio virus. No serological evidence was found linking the neurological disease that affected local equines to WNV. All caimans and sheep were negative by blocking ELISA for flaviviruses. There were no seropositive equines for Bussuquara, Iguape, Yellow fever and all four Dengue virus serotypes. The detection of WNV-seropositive equines in ten ranches and ILHV and SLEV-seropositive equines in fourteen ranches of two different sub-regions of Pantanal is strong evidence of widespread circulation of these flaviviruses in the region.

  2. Serological evidence of widespread circulation of West Nile virus and other flaviviruses in equines of the Pantanal, Brazil.

    Science.gov (United States)

    Pauvolid-Corrêa, Alex; Campos, Zilca; Juliano, Raquel; Velez, Jason; Nogueira, Rita Maria Ribeiro; Komar, Nicholas

    2014-02-01

    A recent study reported neutralizing antibodies to West Nile virus (WNV) in horses from four ranches of southern Pantanal. To extend that study, a serosurvey for WNV and 11 Brazilian flaviviruses was conducted with 760 equines, 238 sheep and 61 caimans from 17 local cattle ranches. Among the tested equines, 32 were collected from a ranch where a neurologic disorder outbreak had been recently reported. The sera were initially screened by using a blocking ELISA and then titrated by 90% plaque-reduction neutralization test (PRNT90) for 12 flaviviruses. Employing the criterion of 4-fold greater titer, 78 (10.3%) equines were seropositive for Ilheus virus, 59 (7.8%) for Saint Louis encephalitis virus, 24 (3.2%) for WNV, two (0.3%) for Cacipacore virus and one (0.1%) for Rocio virus. No serological evidence was found linking the neurological disease that affected local equines to WNV. All caimans and sheep were negative by blocking ELISA for flaviviruses. There were no seropositive equines for Bussuquara, Iguape, Yellow fever and all four Dengue virus serotypes. The detection of WNV-seropositive equines in ten ranches and ILHV and SLEV-seropositive equines in fourteen ranches of two different sub-regions of Pantanal is strong evidence of widespread circulation of these flaviviruses in the region.

  3. Bovine papillomavirus in Brazil: detection of coinfection of unusual types by a PCR-RFLP method.

    Science.gov (United States)

    Carvalho, R F; Sakata, S T; Giovanni, D N S; Mori, E; Brandão, P E; Richtzenhain, L J; Pozzi, C R; Arcaro, J R P; Miranda, M S; Mazzuchelli-de-Souza, J; Melo, T C; Comenale, G; Assaf, S L M R; Beçak, W; Stocco, R C

    2013-01-01

    Bovine papillomavirus (BPV) is recognized as a causal agent of benign and malignant tumors in cattle. Thirteen types of BPV are currently characterized and classified into three distinct genera, associated with different pathological outcomes. The described BPV types as well as other putative ones have been demonstrated by molecular biology methods, mainly by the employment of degenerated PCR primers. Specifically, divergences in the nucleotide sequence of the L1 gene are useful for the identification and classification of new papillomavirus types. On the present work, a method based on the PCR-RFLP technique and DNA sequencing was evaluated as a screening tool, allowing for the detection of two relatively rare types of BPV in lesions samples from a six-year-old Holstein dairy cow, chronically affected with cutaneous papillomatosis. These findings point to the dissemination of BPVs with unclear pathogenic potential, since two relatively rare, new described BPV types, which were first characterized in Japan, were also detected in Brazil.

  4. Test and validation of methods to sample and detect human virus from environmental surfaces using norovirus as a model virus

    DEFF Research Database (Denmark)

    Ibfelt, T.; Frandsen, T.; Permin, Anders

    2016-01-01

    : To validate and test efficient and reliable procedures to detect multiple human pathogenic viruses on surfaces. Methods: The study was divided into two parts. In Part A, six combinations of three different swabs (consisting of cotton, foamed cotton, or polyester head) and two different elution methods (direct...

  5. Detection of sweet potato virus C, sweet potato virus 2 and sweet potato feathery mottle virus in Portugal.

    Science.gov (United States)

    Varanda, Carla M R; Santos, Susana J; Oliveira, Mônica D M; Clara, Maria Ivone E; Félix, Maria Rosário F

    2015-06-01

    Field sweet potato plants showing virus-like symptoms, as stunting, leaf distortion, mosaic and chlorosis, were collected in southwest Portugal and tested for the presence of four potyviruses, sweet potato virus C (SPVC), sweet potato virus 2 (SPV2), sweet potato feathery mottle virus (SPFMV), sweet potato virus G (SPVG), and the crinivirus sweet potato chlorotic stunt virus (SPCSV). DsRNA fractions were extracted from symptomatic leaves and used as templates in single and multiplex RT-PCR assays using previously described specific primers for each analyzed virus. The amplified reaction products for SPVC, SPV2 and SPFMV were of expected size, and direct sequencing of PCR products revealed that they correspond to the coat protein gene (CP) and showed 98%, 99% and 99% identity, respectively, to those viruses. Comparison of the CP genomic and amino acid sequences of the Portuguese viral isolates recovered here with those of ten other sequences of isolates obtained in different countries retrieved from the GenBank showed very few differences. The application of the RT-PCR assays revealed for the first time the presence of SPVC and SPFMV in the sweet potato crop in Portugal, the absence of SPVG and SPCSV in tested plants, as well as the occurrence of triple virus infections under field conditions.

  6. JC virus detection and JC virus-specific immunity in natalizumab-treated Multiple Sclerosis patients

    Directory of Open Access Journals (Sweden)

    Mancuso Roberta

    2012-12-01

    Full Text Available Abstract Background The use of natalizumab in multiple sclerosis (MS may favour JC virus reactivation; this phenomenon is usually asymptomatic but can, albeit rarely, evolve into frank progressive multifocal leucoencephalopathy (PML. Methods JCV-specific CD8+ T lymphocytes were evaluated by flow cytometry over a 24-month period in 24 natalizumab-treated MS patients in whom JCV DNA was or was not detected in blood using quantitative real-time polymerase chain reaction; all these cases were asymptomatic. Results Perforin- and grazymes-containing VP-1-specific CD8+ T lymphocytes were reduced whereas CD107a-expressing cells were increased in JCV positive patients, suggesting an active degranulation of these cells; naïve CD8+ T lymphocytes were also decreased whereas memory cells were increased in patients in whom JCV reactivation was observed. Conclusion The presence of a CD8+ T lymphocyte-mediated effector immune response offers a greater insight into reactivation of JCV and its clinical sequelae, and may help the monitoring of patients on natalizumab therapy.

  7. New influenza A(H7N7) viruses detected in live poultry markets in China.

    Science.gov (United States)

    Cui, Pengfei; Deng, Guohua; Shi, Jianzhong; Kong, Huihui; Liu, Liling; Guan, Yuntao; Suzuki, Yasuo; Chen, Hualan

    2016-12-01

    H7N7 avian influenza viruses have been widely detected in wild birds and domestic poultry since they were first detected in chickens in Italy in 1902. They can occasionally transmit to humans. Here, we isolated six H7N7 viruses in live poultry markets during routine surveillance from 2010 to 2013. Sequences analysis revealed that these viruses are reassortants bearing genes of H3N8, H7N3, H7N7, and H10N7 influenza viruses detected in wild birds and ducks, and can be categorized into three genotypes (A, B, and C). All six viruses bound to both human-type and avian-type receptors. The viruses in genotype B and C could replicate efficiently in the lungs and nasal turbinates of mice without prior adaptation, and the genotype C virus also replicated in the brain of two of three mice tested. It is important to continue to monitor the evolution of H7N7 viruses and to evaluate their potential to cause human infections. Copyright © 2016 Elsevier Inc. All rights reserved.

  8. Host-encoded reporters for the detection and purification of multiple enveloped viruses.

    Science.gov (United States)

    Ketteler, Robin; Tomov, Vesselin; Neunkirchner, Alina; Xie, Qiang; Pickl, Winfried F; Seed, Brian

    2010-08-01

    The identification of host cell factors for virus replication holds great promise for the development of new antiviral therapies. Recently, high-throughput screening methods have emerged as powerful tools to identify candidate host factors for therapeutic intervention. The development of assay systems suitable for large-scale automated screening is of particular importance for novel viruses with high pathogenic potential for which limited biological information can be developed in a short period of time. This report presents a general enzymatic reporter system for the detection and characterization of multiple enveloped viruses that does not rely on engineering of the virus. Instead, reporter enzymes are incorporated into virus particles by targeting to lipid microdomains in producer cells. The approach allows a variety of human pathogenic enveloped viruses to be detected by sensitive, inexpensive and automatable enzymatic assays. Tagged viruses can be purified quickly and efficiently by a magnetic bead-based capture method. The method allows general detection of enveloped viruses without prior reference to their sequence. Copyright (c) 2010 Elsevier B.V. All rights reserved.

  9. [Detection of Aichi virus in stool samples from children in Lanzhou].

    Science.gov (United States)

    Li, Bo-Wen; Liang, Sha; Xie, Jin-Xin; Li, Jie; Lan, Bei; Li, Yong-Qing; Li, Hui-Ying; Cheng, Wei-Xia; Li, Dan-Di; Jin, Yu; Duan, Zhao-Jun

    2010-02-01

    To identification and analysis Aichi virus from diarrhea and normal children in Lanzhou, and discuss the relationship between Aichi virus and Infant Diarrhea. According to the literature published data, Using RT-PCR method to amplified Aichi virus 3CD fragment and the positive products were sequenced and determined, and made the alignment analysis between the nucleotide sequences of the amplified fragment with the known sequence of this virus. There was 1 case detection of Aichi virus in the 46 hospitalized children with diarrhea and 299 children with diarrhea out-patients specifically, Overall detection rate was 0.06%, and there was no Aichi virus was detected in normal control children. 2 viral 3CD gene and the known reference strains of nucleotide sequences were 97%, while phylogenetic analysis showed that genotype of 2 viral belongs to the B. There existed B Genotype of Aichi virus in China, and more research is needed to clarified the etiology and epidemiology of Aichi virus characteristics.

  10. Characterization of Tomato yellow spot virus, a novel tomato-infecting begomovirus in Brazil Caracterização do Tomato yellow spot virus, um novo begomovírus isolado de tomateiro no Brasil

    Directory of Open Access Journals (Sweden)

    Renata Faier Calegario

    2007-09-01

    Full Text Available The objective of this work was the biological and molecular characterization of a begomovirus detected in São Joaquim de Bicas, Minas Gerais, Brazil, named TGV-[Bi2], by determining its host range, complete nucleotide sequence and phylogenetic relationships with other begomoviruses. Biological characterization consisted of a host range study using either sap inoculation or particle bombardment as inoculation methods. The yellow spot virus can infect plants in Solanaceae and Amaranthaceae, including economically importat crops as sweet pepper, and weeds as Datura stramonium and Nicotiana silvestris. For the molecular characterization, the full-length genome (DNA-A and DNA-B was amplified, cloned and completely sequenced. Sequence comparisons and phylogenetic analyses indicated that TGV-[Bi2] constitutes a novel begomovirus species named Tomato yellow spot virus (ToYSV, closely related to Sida mottle virus (SiMoV.O objetivo deste trabalho foi a caracterização biológica e molecular de um begomovírus detectado em tomateiros em São Joaquim de Bicas, Minas Gerais, denominado TGV-[Bi2]. A caracterização biológica consistiu em teste de gama de hospedeiros, realizado por meio de inoculação via extrato foliar tamponado ou bombardeamento de partículas. O isolado TGV-[Bi2] infecta plantas das famílias Solanaceae e Amaranthaceae, inclusive espécies economicamente importantes como o pimentão, e algumas plantas daninhas como Datura stramonium e Nicotiana silvestris. A caracterização molecular consistiu na clonagem e seqüenciamento de seu genoma completo (DNA-A e DNA-B. A comparação de seqüências e análise filogenética indicaram que o TGV-[Bi2] constitui uma nova espécie de begomovírus, denominada Tomato yellow spot virus (ToYSV, filogeneticamente relacionado ao Sida mottle virus (SiMoV.

  11. Respiratory viruses in children with cystic fibrosis: viral detection and clinical findings

    Science.gov (United States)

    Burns, Jane L.; Emerson, Julia; Kuypers, Jane; Campbell, Angela P.; Gibson, Ronald L.; McNamara, Sharon; Worrell, Kelly; Englund, Janet A.

    2011-01-01

    Please cite this paper as: Burns et al. (2011) Respiratory viruses in children with cystic fibrosis: viral detection and clinical findings. Influenza and Other Respiratory Viruses 6(3), 218–223. Background  Viral detection from different respiratory sample types in children with cystic fibrosis (CF) is facilitated by available molecular methods, but optimum sampling strategies have not been identified. In addition, associations between viral detection and respiratory symptoms are not well described. Objectives  Study goals were to compare molecular detection of viruses from concurrent upper airway and sputum samples in children with CF and to describe relative frequency of respiratory viral infections and identify potential clinical associations. Methods  We conducted a 2‐year prospective surveillance study in 44 children with CF aged 6–18 years. Upper airway and sputum samples were collected quarterly and during pulmonary exacerbations and tested for respiratory syncytial virus (RSV), influenza viruses, parainfluenza viruses types 1–4, human metapneumovirus, coronaviruses, rhinoviruses, and adenoviruses. Physical exams and symptom surveys were used to identify respiratory signs and symptoms. Results  Upper airway samples were collected at 359 visits; concordance of PCR‐based viral detection was examined in a subset of paired upper airway and sputum samples from 21 participants at 92 visits. Rhinovirus was the most commonly detected virus (23·1% overall), and rhinovirus detection was the same for both sample types (21·7% each). Sensitivity and specificity for the detection of rhinovirus in sputum relative to upper airway sampling were 70% and 91·7%, respectively. Respiratory symptoms associated with rhinovirus detection included increased cough, increased nasal congestion, increased sputum production, and wheezing. Conclusions  A relatively high frequency of rhinovirus detection was observed by either upper airway or sputum samples, and

  12. Single virus detection by means of atomic force microscopy in combination with advanced image analysis.

    Science.gov (United States)

    Bocklitz, Thomas; Kämmer, Evelyn; Stöckel, Stephan; Cialla-May, Dana; Weber, Karina; Zell, Roland; Deckert, Volker; Popp, Jürgen

    2014-10-01

    In the present contribution virions of five different virus species, namely Varicella-zoster virus, Porcine teschovirus, Tobacco mosaic virus, Coliphage M13 and Enterobacteria phage PsP3, are investigated using atomic force microscopy (AFM). From the resulting height images quantitative features like maximal height, area and volume of the viruses could be extracted and compared to reference values. Subsequently, these features were accompanied by image moments, which quantify the morphology of the virions. Both types of features could be utilized for an automatic discrimination of the five virus species. The accuracy of this classification model was 96.8%. Thus, a virus detection on a single-particle level using AFM images is possible. Due to the application of advanced image analysis the morphology could be quantified and used for further analysis. Here, an automatic recognition by means of a classification model could be achieved in a reliable and objective manner. Copyright © 2014 Elsevier Inc. All rights reserved.

  13. Detection of Aichi virus shedding in a child with enteric and extraintestinal symptoms in Hungary.

    Science.gov (United States)

    Reuter, Gábor; Boldizsár, Akos; Papp, Gábor; Pankovics, Péter

    2009-01-01

    Aichi virus, genus Kobuvirus, family Picornaviridae, has been proposed as a causative agent of gastroenteritis in human. Although high seroprevalence has been detected, it has been identified in only a few cases. We report detection of Aichi virus in Hungary. A total of 65 stool samples were tested retrospectively, collected from children with diarrhea, by reverse transcription-polymerase chain reaction. One (1.5%) sample from a 3-year-old girl was positive. Besides diarrhea, fever, purulent conjunctivitis and respiratory symptoms were also present at the same time with virus shedding. The genotype A virus, Kobuvirus/human/Szigetvar-HUN298/2000/Hungary (FJ225407), has 96% nucleotide identity to Aichi virus.

  14. Nested PCR for Rapid Detection of Mumps Virus in Cerebrospinal Fluid from Patients with Neurological Diseases

    OpenAIRE

    Poggio, Gustavo Palacios; Rodriguez, Claudia; Cisterna, Daniel; Freire, María Cecilia; Cello, Jerónimo

    2000-01-01

    In this study, we have developed a reverse transcription (RT)-nested polymerase chain reaction (n-PCR) for the detection of mumps virus RNA in cerebrospinal fluid (CSF) from patients with neurological infections. A specific 112-bp fragment was amplified by this method with primers from the nucleoprotein of the mumps virus genome. The mumps virus RT–n-PCR was capable of detecting 0.001 PFU/ml and 0.005 50% tissue culture infective dose/ml. This method was found to be specific, since no PCR pro...

  15. A duplex PCR assay for simultaneous detection and differentiation of Capripoxvirus and Orf virus.

    Science.gov (United States)

    Zheng, Min; Liu, Qi; Jin, Ningyi; Guo, Jiangang; Huang, Xia; Li, Huaming; Zhu, Wei; Xiong, Yi

    2007-08-01

    A duplex polymerase chain reaction (PCR) was developed and optimized for simultaneous detection and differentiation of Capripoxvirus (CPV) (including goat pox virus and sheep pox virus) and orf virus (ORFV). Two sets of specific oligonucleotide primers were designed and used for CPV and ORFV, respectively. The duplex PCR DNA products, which consisted of fragments of 413 bp for A29L gene of CPV, and 708 bp for P55 gene of ORFV, were visualized by gel electrophoresis. The developed assay was found to be highly specific and sensitive with a detection limit of 1 plaque forming unit (pfu) for both CPV and CPV.

  16. Selected mild strains of Passion fruit woodiness virus (PWV fail to protect pre-immunized vines in Brazil

    Directory of Open Access Journals (Sweden)

    Novaes Quelmo Silva de

    2003-01-01

    Full Text Available The Passion fruit woodiness virus (PWV is the most important virus affecting passion fruit (Passiflora edulis f. flavicarpa Deg. crops in Brazil. The main purpose of this work was to select mild strains of PWV and to evaluate their protective effect against a severe strain of the virus. Three mild strains were selected from outstanding plants found in orchards severely affected by the virus (F-101, F-102 and F-103 and three others were obtained from blisters formed in passion fruit vine leaves showing mosaic (F-99, F-144 and F-145. The protective effect of the mild strains was evaluated for vines under greenhouse and field conditions. Plants pre-immunized with mild strains F-101, F-102 and F-144, in a greenhouse, had partial protection against the severe strain PWV-SP. In a first field experiment, all passion fruit vines pre-immunized with the six selected mild strains showed severe symptoms of the disease, approximately four months after the challenge inoculation with the PWV-SP strain. Results from a second field experiment, with vines pre-immunized with strains F-101 and F-144, followed by a quantitative evaluation of the mild strains in different leaves of the protected plants, indicated that breakdown in protection seems to be related to the low concentration and/or irregular distribution of the mild strains in leaves, which allows the existence of infection sites available for the establishment of the severe strain. Pre-immunization was not an appropriate alternative for the control of the passion fruit woodiness disease.

  17. Oligonucleotide microarray-based detection and genotyping of Plum pox virus.

    Science.gov (United States)

    Pasquini, Graziella; Barba, Marina; Hadidi, Ahmed; Faggioli, Francesco; Negri, Rodolfo; Sobol, Iris; Tiberini, Antonio; Caglayan, Kadriye; Mazyad, Hamed; Anfoka, Ghandi; Ghanim, Murad; Zeidan, Mohammad; Czosnek, Henryk

    2008-01-01

    Plum pox virus (PPV) is the most damaging viral pathogen of stone fruits. The detection and identification of its strains are therefore of critical importance to plant quarantine and certification programs. Existing methods to screen strains of PPV suffer from significant limitations such as the simultaneous detection and genotyping of several strains of PPV in samples infected with different isolates of the virus. A genomic strategy for PPV screening based on the viral nucleotide sequence was developed to enable the detection and genotyping of the virus from infected plant tissue or biological samples. The basis of this approach is a long 70-mer oligonucleotide DNA microarray capable of simultaneously detecting and genotyping PPV strains. Several 70-mer oligonucleotide probes were specific for the detection and genotyping of individual PPV isolates to their strains. Other probes were specific for the detection and identification of two or three PPV strains. One probe (universal), derived from the genome highly conserved 3' non-translated region, detected all individual strains of PPV. This universal PPV probe, combined with probes specific for each known strain, could be used for new PPV strain discovery. Finally, indirect fluorescent labeling of cDNA with cyanine after cDNA synthesis enhanced the sensitivity of the virus detection without the use of the PCR amplification step. The PPV microarray detected and identified efficiently the PPV strains in PPV-infected peach, apricot and Nicotiana benthamiana leaves. This PPV detection method is versatile, and enables the simultaneous detection of plant pathogens.

  18. Detection of West Nile Virus and other common equine viruses in three locations from the Leeward Islands, West Indies.

    Science.gov (United States)

    Bolfa, Pompei; Jeon, Isaac; Loftis, Amanda; Leslie, Teresa; Marchi, Silvia; Sithole, Fortune; Beck, Cecile; Lecollinet, Sylvie; Zientara, Stephan; Hans, Aymeric; Issel, Charles J

    2017-10-01

    Equines in the West Indies are used for recreational purposes, tourism industry, racing and agriculture or can be found in feral populations. Little is known in the Caribbean basin about the prevalence of some major equine infectious diseases, some with zoonotic potential, listed as reportable by the OIE. Our objective was to study the prevalence of antibodies for West Nile Virus (WNV), Equine Herpes Virus-1 and 4 (EHV-1 and EHV-4), Equine Influenza (EI), Equine Viral Arteritis (EVA) and Equine Infectious Anemia Virus (EIAV) using a retrospective serological convenience study. We used 180 equine serum samples, 140 from horses and 40 from donkeys in St. Kitts, Nevis, and Sint Eustatius, collected between 2006 and 2015 that were tested with ELISA kits and virus neutralization (for WNV and EVA). Combining ELISA with virus neutralization testing, 25 (13.8%) equine sera were WNV positive (a mixture of indigenous and imported equines) and 3 sera (1.6%) showed doubtful results. For EHV-1, 41 equines (23.7%), mean age 6.7 years, were seropositive. For EHV-4, 138 equines were found seropositive (82.8%), mean age 6.3 years. For EI, 49 equines (27.2%), mean age 7.5 years, were seropositive on ELISA, some previously vaccinated horses. No antibodies against EAV were found on virus neutralization testing, although one animal (0.6%), was EAV positive on ELISA. All samples were EIAV negative. The seroprevalence for EHV-1 and EHV-4 is similar to other parts of the world. For the first time in the study location serologic evidence of antibodies against WNV and EI is reported. This was found in both indigenous and imported animals, highlighting the need for developing proper surveillance plans based on complementary methods of virus detection. Further studies will be needed to define the prevalence, rates of transmission, characterize local virus strains, and study their impact on these populations. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. Primeiro relato do Tomato spotted wilt virus (TSWV em Campanula medium L. no Brasil First report of Tomato spotted wilt virus (TSWV infecting Campanula medium in Brazil

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    Ricardo Gioria

    2010-06-01

    Full Text Available Plantas de campânula (Campanula medium exibindo mosaico e necrose foliar e anéis em flores foram coletadas em uma estufa comercial de flores na região de Atibaia, SP. Suspeitando de possível etiologia viral, amostras de tecido lesionado foram analisadas por ensaios de transmissão mecânica, microscopia eletrônica e sorologia. Todos os resultados apontaram para a presença do Tomato spotted wilt virus (TSWV como o responsável pelos sintomas. Esse é o primeiro relato deste patógeno em campânula no Brasil.Plants of bellflower (Campanula medium exhibiting symptoms of mosaic, leaf necrosis and flower ring spot were found in a commercial crop in a greenhouse in Atibaia, São Paulo State. Electron microscopy, serology and biological assays indicated the presence of Tomato spotted wilt virus (TSWV. This is the first report of this pathogen in bellflower plants in Brazil.

  20. Epidemiology of hepatitis B virus infection in first-time blood donors in the southwestern region of Goiás, central Brazil

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    Giulena Rosa Leite Cardoso dos Anjos

    2011-02-01

    Full Text Available INTRODUCTION: Little is known about the epidemiology of hepatitis B virus (HBV infection in populations from inner cities, especially in Central Brazil. Thus the objective of this study was to estimate the prevalence of HBV infection, and to analyze the factors associated with HBV infection, in a population of first-time blood donors in the southwestern region of Goiás, Central Brazil. METHODS: A total of 984 individuals were interviewed and gave blood samples to detect serological markers of HBV (HBsAg, anti-HBs, and anti-HBc by enzyme linked immunosorbent assays. RESULTS: An overall prevalence of 6.9% was found for HBV, with constituent prevalence rates of 3.6% and 11.6%, in subjects classified as fit and unfit to donate blood according the epidemiological screening, respectively. Only three individuals were positive for anti-HBs alone, suggesting previous vaccination against HBV. The variables of prior blood transfusion (OR = 2.3, tattoo/piercing (OR = 2.1, illicit drug use (OR = 2.3, sex with a partner with hepatitis (OR = 14.7, and history of sexually transmitted diseases (OR = 2.9 were independently associated with HBV-positivity. These data suggested a low endemicity of hepatitis B in the studied population. CONCLUSION: The findings of low hepatitis B immunization coverage and the association of hepatitis B with risky behavior highlight that there is a need to intensify hepatitis B prevention programs in the southwest region of Goiás.

  1. Rapid quantitative PCR assays for the simultaneous detection of herpes simplex virus, varicella zoster virus, cytomegalovirus, Epstein-Barr virus, and human herpesvirus 6 DNA in blood and other clinical specimens

    NARCIS (Netherlands)

    Engelmann, I.; Petzold, D. R.; Kosinska, A.; Hepkema, B. G.; Schulz, T. F.; Heim, A.

    Rapid diagnosis of human herpesvirus primary infections or reactivations is facilitated by quantitative PCRs. Quantitative PCR assays with a standard thermal cycling profile permitting simultaneous detection of herpes simplex virus (HSV), varicella zoster virus (VZV), cytomegalovirus (CMV),

  2. Molecular Detection and Characterization of Gastroenteritis Viruses Occurring Naturally in the Stream Waters of Manaus, Central Amazônia, Brazil▿

    Science.gov (United States)

    Miagostovich, Marize P.; Ferreira, Fabiana F. M.; Guimarães, Flávia R.; Fumian, Túlio M.; Diniz-Mendes, Leonardo; Luz, Sérgio Luiz B.; Silva, Luciete A.; Leite, José Paulo G.

    2008-01-01

    To assess the presence of the four main viruses responsible for human acute gastroenteritis in a hydrographic network impacted by a disordered urbanization process, a 1-year study was performed involving water sample collection from streams in the hydrographic basin surrounding the city of Manaus, Amazonas, Brazil. Thirteen surface water sample collection sites, including different areas of human settlement characterized as urban, rural, and primary forest, located in the Tarumã-Açu, São Raimundo, Educandos, and Puraquequara microbasins, were defined with a global positioning system. At least one virus was detected in 59.6% (31/52) of the water samples analyzed, and rotavirus was the most frequent (44.2%), followed by human adenovirus (30.8%), human astrovirus (15.4%), and norovirus (5.8%). The viral contamination observed mainly in the urban streams reflected the presence of a local high-density population and indicated the gastroenteritis burden from pathogenic viruses in the water, principally due to recreational activities such as bathing. The presence of viral genomes in areas where fecal contamination was not demonstrated by bacterial indicators suggests prolonged virus persistence in aquatic environments and emphasizes the enteric virus group as the most reliable for environmental monitoring. PMID:18065620

  3. Detection of viruses in olive trees in Croatian Istria

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    Marta LUIGI

    2011-05-01

    Full Text Available Normal 0 14 false false false IT ZH-TW X-NONE MicrosoftInternetExplorer4 /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Tabella normale"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0cm 5.4pt 0cm 5.4pt; mso-para-margin-top:0cm; mso-para-margin-right:0cm; mso-para-margin-bottom:10.0pt; mso-para-margin-left:0cm; line-height:115%; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:"Times New Roman"; mso-bidi-theme-font:minor-bidi;} Following identification of four viruses in a general survey of olive trees throughout Croatia, a detailed survey was conducted in 2009 in the field collection of the Institute of Agriculture and Tourism in Poreč (an important reservoir of Istrian native olive germplasm in order to evaluate the sanitary status of the most important Croatian Istria olive cultivars. Twenty five samples from symptomatic or symptomless trees were collected from five autochthonous and four exotic cultivars. All the samples were tested by RT-PCR for the presence of: Olive leaf yellowing associated virus (OLYaV, Cherry leaf roll virus (CLRV, Strawberry latent ring spot virus (SLRSV, Arabis mosaic virus (ArMV, Olive latent virus-1 (OLV-1, Cucumber mosaic virus (CMV, Olive latent virus-2 (OLV-2 and Tobacco necrosis virus D (TNV-D. Six of the 25 plants were found positive to CLRV; all infected plants showed leaf and fruit deformation and leaf yellowing. Four positive samples were from the native cv. Buža whereas the other two were from two exotic cultivars: Ascolana tenera and Frantoio. The presence of CLRV,  either in native or imported plants, highlights the importance of strict phytosanitary regulations to prevent incursion of key

  4. Detection of West Nile virus and tick-borne encephalitis virus in birds in Slovakia, using a universal primer set.

    Science.gov (United States)

    Csank, Tomáš; Bhide, Katarína; Bencúrová, Elena; Dolinská, Saskia; Drzewnioková, Petra; Major, Peter; Korytár, Ľuboš; Bocková, Eva; Bhide, Mangesh; Pistl, Juraj

    2016-06-01

    West Nile virus (WNV) is a mosquito-borne neurotropic pathogen that presents a major public health concern. Information on WNV prevalence and circulation in Slovakia is insufficient. Oral and cloacal swabs and bird brain samples were tested for flavivirus RNA by RT-PCR using newly designed generic primers. The species designation was confirmed by sequencing. WNV was detected in swab and brain samples, whereas one brain sample was positive for tick-borne encephalitis virus (TBEV). The WNV sequences clustered with lineages 1 and 2. These results confirm the circulation of WNV in birds in Slovakia and emphasize the risk of infection of humans and horses.

  5. Detection and molecular characterization of Babesia caballi and Theileria equi isolates from endemic areas of Brazil.

    Science.gov (United States)

    Heim, Alexandra; Passos, Lygia M F; Ribeiro, Múcio F B; Costa-Júnior, Lívio M; Bastos, Camila V; Cabral, Dagmar D; Hirzmann, Jörg; Pfister, Kurt

    2007-12-01

    Blood samples were collected from 487 adult horses, including 83 pregnant mares, at a slaughterhouse located in Araguari, Minas Gerais State, Brazil. For each blood sample, the packed cell volume (PCV) was determined, and Giemsa-stained smears were microscopically examined for the presence of hemoparasites. The plasma was examined by the indirect fluorescent antibody test for detection of antibodies against Babesia caballi and Theileria equi. In addition, DNA was extracted and analyzed by a multiplex real-time polymerase chain reaction (PCR), specific for B. caballi and T. equi. Products of PCR were sequenced and compared with each other and with known sequences. The serological results showed a total prevalence of 91.0% for T. equi and 83.0% for B. caballi, while by PCR, prevalences of 59.7% for T. equi and 12.5% for B. caballi were observed. However, no correlations were seen between positivity (neither by serology nor by PCR) and PCV values. As expected, the microscopic examination of blood smears showed low sensitivity in detecting the infections when compared to the PCR. Only 35 out of 570 blood smears were positive, with parasitemias below 0.1%. No congenital transmission was detectable. As far as sequencing is concerned, no differences were seen among the isolates of each species nor among them and known sequences available. These results confirm, by molecular methods, the high prevalence rates of T. equi and B. caballi infections in carrier horses in Brazil. However, no diversity was observed among the isolates within the studied regions.

  6. Molecular detection of virulence factors among food and clinical Enterococcus faecalis strains in South Brazil

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    A.W. Medeiros

    2014-01-01

    Full Text Available The present report aimed to perform a molecular epidemiological survey by investigating the presence of virulence factors in E. faecalis isolated from different human clinical (n = 57 and food samples (n = 55 in Porto Alegre, Brazil, collected from 2006 to 2009. In addition, the ability to form biofilm in vitro on polystyrene and the β-haemolytic and gelatinase activities were determined. Clinical strains presented a higher prevalence of aggregation substance (agg, enterococcal surface protein (esp and cytolysin (cylA genes when compared with food isolates. The esp gene was found only in clinical strains. On the other hand, the gelatinase (gelE and adherence factor (ace genes had similar prevalence among the strains, showing the widespread occurrence of these virulence factors among food and clinical E. faecalis strains in South Brazil. More than three virulence factor genes were detected in 77.2% and 18.2% of clinical and food strains, respectively. Gelatinase and β-haemolysin activities were not associated with the presence of gelE and cylA genes. The ability to produce biofilm was detected in 100% of clinical and 94.6% of food isolates, and clinical strains were more able to form biofilm than the food isolates (Student's t-test, p < 0.01. Results from the statistical analysis showed significant associations between strong biofilm formation and ace (p = 0.015 and gelE (p = 0.007 genes in clinical strains. In conclusion, our data indicate that E. faecalis strains isolated from clinical and food samples possess distinctive patterns of virulence factors, with a larger number of genes that encode virulence factors detected in clinical strains.

  7. A case-control study on the association of hepatitis B virus infection and hepatocellular carcinoma in Northeast Brazil

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    Cotrim Helma

    1992-01-01

    Full Text Available Hepatitis B virus (HBV serological markers were investigated in 40 incident cases of hepatocellular carcinoma (HCC and in two age and sex matched control groups, comprising 40 patients with other cancers and 80 healthy individuals, resident in Bahia, Brazil. Serologic tests were done by radioimmunoassay. The study observed high proportion of seropositivity to HBsAg (42.5% and of those presenting HBsAg or antiHBc (65.0% among HCC cases, higher in men than women and in those aged 17 to 30 years old. HBsAg seropositivity among HCC patients was greater than in the control group with other cancers (7.5% and in healthy controls (2.5%, corresponding to odds ratio estimates of 15.0 (95% CI 3.29, 68.30 and 33.0 (95% CI 9.13, 119.28, both statistically significant. HBeAg was not observed and antiHBe was present in 41.2% of cases, suggesting the absence of viral replication, possibly with viral DNA intergration into the hepatocyte genome. The presence of cirrhosis was associated with HBsAg seropositivity among HCC cases. A history of chronic alcoholism is shown to be more frequently related to those cases with cirrhosis. This study highlights the relevant association between HCC and HBV in Northeast Brazil, particularly for young individuals, and the high risk of development of HCC for HBsAg carriers.

  8. A case-control study on the association of hepatitis B virus infection and hepatocellular carcinoma in Northeast Brazil

    Directory of Open Access Journals (Sweden)

    Helma Cotrim

    1992-10-01

    Full Text Available Hepatitis B virus (HBV serological markers were investigated in 40 incident cases of hepatocellular carcinoma (HCC and in two age and sex matched control groups, comprising 40 patients with other cancers and 80 healthy individuals, resident in Bahia, Brazil. Serologic tests were done by radioimmunoassay. The study observed high proportion of seropositivity to HBsAg (42.5% and of those presenting HBsAg or antiHBc (65.0% among HCC cases, higher in men than women and in those aged 17 to 30 years old. HBsAg seropositivity among HCC patients was greater than in the control group with other cancers (7.5% and in healthy controls (2.5%, corresponding to odds ratio estimates of 15.0 (95% CI 3.29, 68.30 and 33.0 (95% CI 9.13, 119.28, both statistically significant. HBeAg was not observed and antiHBe was present in 41.2% of cases, suggesting the absence of viral replication, possibly with viral DNA intergration into the hepatocyte genome. The presence of cirrhosis was associated with HBsAg seropositivity among HCC cases. A history of chronic alcoholism is shown to be more frequently related to those cases with cirrhosis. This study highlights the relevant association between HCC and HBV in Northeast Brazil, particularly for young individuals, and the high risk of development of HCC for HBsAg carriers.

  9. Prevalence of human papillomavirus and Epstein-Barr virus DNA in penile cancer cases from Brazil

    Directory of Open Access Journals (Sweden)

    Larissa Alves Afonso

    2012-02-01

    Full Text Available Penile cancer is a potentially mutilating disease. Although its occurrence is relatively rare worldwide, penile cancer rates can be high in developing countries. A few studies have been conducted on the involvement of human papillomavirus (HPV in penile carcinoma, which have found HPV present in 30-70% of penile malignant lesions, with a higher prevalence of HPV 16 and 18. It has been assumed that cofactors, such as Epstein-Barr virus (EBV infections, may play a role in the progression of penile neoplasia. The aim of this study was to determine HPV and EBV prevalence in 135 penile malignant lesions from Brazilian men through the use of MY09/11 polymerase chain reaction (PCR, type-specific PCR and restriction fragment length polymorphism analysis. HPV prevalence among the men tested was 60.7%. Of the men who tested positive, 27 presented with HPV 16 (29.7%, five with HPV 18 (5.5%, 21 with HPV 45 (23.1% and nine with HPV 6 (9.9%. Seven mixed infections were detected (9.2%, while 11 cases remained untyped (13.4%. Regarding EBV positivity, 46.7% of the samples contained EBV DNA with EBV-1 as the most prevalent type (74.6%. More than 23% of the men were co-infected with both HPV and EBV, while 35% presented exclusively with HPV DNA and 20% presented only with EBV DNA. Penile carcinoma aetiology has not been fully elucidated and the role of HPV and EBV infections individually or synergistically is still controversial. Hence, more studies are needed to determine their possible role in carcinogenesis.

  10. No detection of xenotropic murine leukemia virus-related viruses in prostate cancer in Sanandaj, west of Iran.

    Science.gov (United States)

    Khodabandehloo, Mazaher; Hosseini, Weria; Rahmani, Mohammad-Reza; Rezaee, Mohammad-Ali; Hakhamaneshi, Mohammad-Saied; Nikkhoo, Bahram; Jalili, Ali

    2013-01-01

    Multiple etiologies have been hypothesized for prostate cancer, including genetic defects and infectious agents. A recently reported gamaretrovirus, xenotropic murine leukemia virus-related virus (XMRV) has been reported to be detected in prostate cancer. However, this virus has not been detected in similar groups of patients in other studies. Herein, we sought to detect XMRV in prostate cancers and benign controls in Sanandaj, west of Iran. In a case-control study, genomic DNA was extracted from formalin fixed and paraffin embedded prostate tissues from a total of 163 Iranian patients. We developed a conventional and a nested PCR assay using primers targeting to an env specific sequence of XMRV. PCR assays were carried out on 63 prostate cancers and 100 benign prostate hyperplasias. Beta-actin sequences were successfully detected in the DNA extracts from all prostate tissues, confirming DNA extraction integrity. We did not detect XMRV in samples either from prostate cancers or benign prostate hyperplasias using XMRV specific primers. We conclude that in our population XMRV does not play a role in genesis of prostate cancer.

  11. A bead-based multiplex assay for the detection of DNA viruses infecting laboratory rodents.

    Directory of Open Access Journals (Sweden)

    Daniela Höfler

    Full Text Available The Federation of European Laboratory Animal Science Association (FELASA recommends screening of laboratory rodents and biological materials for a broad variety of bacterial agents, viruses, and parasites. Methods commonly used to date for pathogen detection are neither cost-effective nor time- and animal-efficient or uniform. However, an infection even if silent alters experimental results through changing the animals' physiology and increases inter-individual variability. As a consequence higher numbers of animals and experiments are needed for valid and significant results. We developed a novel high-throughput multiplex assay, called rodent DNA virus finder (rDVF for the simultaneous identification of 24 DNA viruses infecting mice and rats. We detected all 24 DNA viruses with high specificity and reproducibility. Detection limits for the different DNA viruses varied between 10 and 1000 copies per PCR. The validation of rDVF was done with DNA isolated from homogenised organs amplified by pathogen specific primers in one multiplex PCR. The biotinylated amplicons were detected via hybridisation to specific oligonucleotide probes coupled to spectrally distinct sets of fluorescent Luminex beads. In conclusion, rDVF may have the potential to replace conventional testing and may simplify and improve routine detection of DNA viruses infecting rodents.

  12. A bead-based multiplex assay for the detection of DNA viruses infecting laboratory rodents.

    Science.gov (United States)

    Höfler, Daniela; Nicklas, Werner; Mauter, Petra; Pawlita, Michael; Schmitt, Markus

    2014-01-01

    The Federation of European Laboratory Animal Science Association (FELASA) recommends screening of laboratory rodents and biological materials for a broad variety of bacterial agents, viruses, and parasites. Methods commonly used to date for pathogen detection are neither cost-effective nor time- and animal-efficient or uniform. However, an infection even if silent alters experimental results through changing the animals' physiology and increases inter-individual variability. As a consequence higher numbers of animals and experiments are needed for valid and significant results. We developed a novel high-throughput multiplex assay, called rodent DNA virus finder (rDVF) for the simultaneous identification of 24 DNA viruses infecting mice and rats. We detected all 24 DNA viruses with high specificity and reproducibility. Detection limits for the different DNA viruses varied between 10 and 1000 copies per PCR. The validation of rDVF was done with DNA isolated from homogenised organs amplified by pathogen specific primers in one multiplex PCR. The biotinylated amplicons were detected via hybridisation to specific oligonucleotide probes coupled to spectrally distinct sets of fluorescent Luminex beads. In conclusion, rDVF may have the potential to replace conventional testing and may simplify and improve routine detection of DNA viruses infecting rodents.

  13. Development of a reverse transcription polymerase chain reaction method for yellow fever virus detection.

    Science.gov (United States)

    Méndez, María C; Domingo, Cristina; Tenorio, Antonio; Pardo, Lissethe C; Rey, Gloria J; Méndez, Jairo A

    2013-09-01

    Yellow fever is considered a re-emerging disease and is endemic in tropical regions of Africa and South America. At present, there are no standardized or commercialized kits available for yellow fever virus detection. Therefore, diagnosis must be made by time-consuming routine techniques, and sometimes, the virus or its proteins are not detected. Furthermore, co-circulation with other flaviviruses, including dengue virus, increases the difficulty of diagnosis. To develop a specific reverse transcriptase-polymerase chain reaction (RT-PCR) and nested PCR-based assay to improve the detection and diagnosis of yellow fever virus using both serum and fresh tissue samples. RT-PCR primers were designed to amplify a short fragment of all yellow fever virus genotypes reported. A second set of primers was used in a nested PCR to increase sensitivity. Thirty-three clinical samples were tested with the standardized reaction. The expected amplicon was obtained in 25 out of 33 samples analyzed using this approach, and 2 more samples tested positive after a subsequent nested PCR approach. This improved technique not only ensures the specific detection of a wide range of yellow fever virus genotypes but also may increase the sensitivity of detection by introducing a second round of amplification, allowing a rapid differential diagnosis between dengue and yellow fever infection, which is required for effective surveillance and opportune epidemiologic measures.

  14. Molecular and serological detection of Leishmania spp. in captive wild animals from Ilha Solteira, SP, Brazil.

    Science.gov (United States)

    Jusi, Márcia Mariza Gomes; Starke-Buzetti, Wilma Aparecida; Oliveira, Trícia Maria Ferreira de Sousa; Tenório, Michely da Silva; Sousa, Lúcio de Oliveira de; Machado, Rosangela Zacarias

    2011-01-01

    Leishmaniasis is a zoonotic disease that affects 12 million people worldwide. Several mammalian species can serve as a reservoir for this disease. Dogs are the main reservoir for visceral leishmaniasis in urban areas, which has become a serious public health concern in Brazil. The aim of this study was to evaluate the presence of Leishmania spp. in captive wild animals from Ilha Solteira, São Paulo, Brazil. Blood and various tissues samples were collected from animals of five different species: Speothos venaticus, Chrysocyon brachyurus, Cerdocyon thous, Pseudalopex vetulus, and Procyon cancrivorus. Antibodies against Leishmania spp. were detected in three wild canids by indirect fluorescent antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA). PCR analyses of blood and bone marrow from all animals were negative, but Leishmania DNA was found in the tissues and skin of seropositive animals. Positive PCR samples were also positive for Leishmania donovani complex. Analysis of sequenced PCR products showed similarities with different regions of Leishmania (Leishmania) infantum and Leishmania (Leishmania) chagasi kinetoplastids. Measures to control visceral leishmaniasis in wild animals kept in Brazilian zoos should be established, as no disease control programs are currently available.

  15. Evaluation of rapid tests for anti-HIV detection in Brazil.

    Science.gov (United States)

    Ferreira Junior, Orlando C; Ferreira, Cristine; Riedel, Maristela; Widolin, Marcya Regina Visinoni; Barbosa-Júnior, Aristides

    2005-10-01

    This assessment in Brazil was to evaluate the performance of commercially available HIV rapid test (RT) against the gold standard testing and to establish a highly sensitive and specific RT algorithm for HIV diagnosis. A prospective, anonymous and unlinked study. An evaluation of seven commercially available RT to compare their performance against the gold standard tests for Brazil. This includes two competing enzyme immunoassays plus a Western blot for confirmation. After informed consent, whole blood samples were collected from volunteers in voluntary counselling and testing sites (n = 400), antenatal clinics (n = 500) and from HIV-positive controls in AIDS treatment centres (n = 200). Two seroconversion panels, one HIV-1 subtype (B, B', C and F) panel and an operational assay performance evaluation were also part of the study parameters. For the seven RT the clinical sensitivity ranged from 97.74 to 100% and clinical specificity from 99.43 to 100%. However, only four RT were considered acceptable after full evaluation. The two EIA had a clinical sensitivity of 100% and clinical specificity of 99.32 and 99.66%. Two RT had the same performance on the seroconversions panels as the EIA. The operational assay performance evaluation for the RT indicated that Hexagon and Capillus could not be classified as simple assays. We have provided evidence that RT assays can perform equally or better than EIA for the detection of HIV antibodies. The simplicity and rapidity of the RT warrants its utilization in an algorithm for a rapid diagnosis of HIV infection.

  16. Molecular and serological detection of Theileria equi and Babesia caballi in donkeys (Equus asinus) in Brazil.

    Science.gov (United States)

    Machado, R Z; Toledo, C Z P; Teixeira, M C A; André, M R; Freschi, C R; Sampaio, P H

    2012-05-25

    Piroplasmosis in donkeys has been recognized as a serious problem of major economic importance, since the affected animals manifest loss of appetite and decreased working capacity. The present work is aimed at detecting infection or exposure of donkeys in São Paulo, Brazil to Theileria (T.) equi and Babesia (B.) caballi using molecular and serological approaches. EDTA-blood and serum samples were collected from 88 donkeys (Equus asinus). From 88 sampled donkeys, 65 (73.86%; 95% confidence interval, PI=63.41, 82.65) and 82 (93.2%; 95% confidence interval, PI=85.75, 97.46) animals showed IgG antibodies to T. equi (by ELISA) and B. caballi (by IFAT), respectively. Twenty-eight (31.81%; 95% confidence interval, PI=22.3, 42.61) and 18 (20.45%; 95% confidence interval, PI=12.6, 30.39) donkeys were positive to T. equi and B. caballi nested PCR assays, respectively. The results indicated that T. equi and B. caballi are prevalent among donkeys in Brazil. Copyright © 2011 Elsevier B.V. All rights reserved.

  17. Detection of viruses in drinking water by concentration on magnetic iron oxide.

    Science.gov (United States)

    Rao, V C; Waghmare, S V; Lakhe, S B

    1981-09-01

    Discharge of raw domestic wastes containing human enteric viruses into water courses, consumption of untreated water from canals, streams, and shallow wells in villages, and cross-contamination of water in the distribution system because of intermittent water supply in urban areas continue to cause widespread outbreaks of infectious hepatitis in India. To detect a low number of viruses in 50- to 100-liter samples of water, a method was developed with magnetic iron oxide as the virus adsorbent. Poliovirus-seeded dechlorinated tap water, adjusted to pH 3.0 and 0.0005 M AlCl3, was filtered through a 10-g bed of iron oxide sandwiched between two AP20 prefilter pads held in a 142-mm-diameter, stainless-steel holder. Virus was eluted from iron oxide by recirculating three times a 100-ml volume of 3% beef extract, pH 9.0. The eluate was reconcentrated to 5 ml by adjusting to pH 3, adding 1 g of iron oxide, stirring for 30 min, and eluting the readsorbed virus with 5 ml of beef extract, pH 9.0. Virus recovery varied from 60 to 80%. Using the above method, we took a survey of drinking water at three locations in Nagpur during 1976 and found the presence of virus in 7 of 50 samples. The quantity of virus recovered ranged from 1 to 7 plaque-forming units per 30 to 60 liters. Virus was detected in some samples even with residual chlorine. No coliforms were detected in the virus-positive samples.

  18. Hepatitis G virus / GB virus C in Brazil. Preliminary report Vírus da Hepatite G / Vírus GB-C no Brasil

    Directory of Open Access Journals (Sweden)

    J.R.R. Pinho

    1996-06-01

    Full Text Available Hepatitis G virus/ GB virus C is a novel flavivirus recently detected in hepatitis non A-E cases. In this study, the presence of this virus in chronic non-B, non-C hepatitis patients was evaluated using GBV-C specific PCR and this virus was detected in one out of thirteen patients. This patient has presented a severe liver failure, has lived for a long time in the Western Amazon basin and no other cause for this clinical picture was reported. The impact of the discovery of this new agent is still under evaluation throughout the world. The study of the prevalence of this virus among chronic hepatitis patients and healthy individuals (as blood donors will furnish subside to evaluate its real pathogenicity.O vírus da Hepatite G ou vírus GB-C é um novo vírus recentemente descoberto em casos de hepatites não A-E. Neste estudo, casos de hepatite crônica não-B, não-C foram testados com uma reação de amplificação específica para GBV-C. Este vírus foi detectado em 1 entre os 13 casos estudados. Este paciente apresentava insuficiência hepática severa, tinha habitado por vários anos na Amazônia Ocidental e nenhuma outra causa para este quadro clínico havia sido relatada. O impacto da descoberta deste novo agente está ainda sendo investigado. O estudo da prevalência deste vírus entre pacientes com hepatite crônica e entre indivíduos sadios (como, por exemplo, doadores de sangue fornecerá subsídios para o estabelecimento de sua real patogenicidade.

  19. Detection and management of antiviral resistance for influenza viruses.

    Science.gov (United States)

    Boivin, Guy

    2013-11-01

    Neuraminidase inhibitors (NAIs) are first-line agents for the treatment and prevention of influenza virus infections. As for other antivirals, the development of resistance to NAIs has become an important concern particularly in the case of A(H1N1) viruses and oseltamivir. The most frequently reported change conferring oseltamivir resistance in that viral context is the H275Y neuraminidase mutation (N1 numbering). Recent studies have shown that, in the presence of the appropriate permissive mutations, the H275Y variant can retain virulence and transmissibility in some viral backgrounds. Most oseltamivir-resistant influenza A virus infections can be managed with the use of inhaled or intravenous zanamivir, another NAI. New NAI compounds and non-neuraminidase agents as well as combination therapies are currently in clinical evaluation for the treatment for severe influenza infections. © 2013 Blackwell Publishing Ltd.

  20. PREVALENCE OF HERPES SIMPLEX VIRUS TYPE 2 AND RISK FACTORS ASSOCIATED WITH THIS INFECTION IN WOMEN IN SOUTHERN BRAZIL

    Science.gov (United States)

    Caldeira, Thaís Duquia Moraes; Gonçalves, Carla Vitola; de Oliveira, Gisele Rodrigues; da Fonseca, Tânia Vieira; Gonçalves, Regina; do Amaral, Clair Teixeira; da Hora, Vanusa Pousada; de Martinez, Ana Maria Barral

    2013-01-01

    SUMMARY The herpes simplex virus type 2 (HVS-2) is the most prevalent infection worldwide. It is a cofactor in the acquisition of human immunodeficiency virus (HIV) and the persistence of human papillomavirus (HPV). This study evaluated the prevalence of HSV-2, using the polymerase chain reaction (PCR), and associated factors in patients treated at the Federal University of Rio Grande (FURG) and Basic Health Units (BHU) in Rio Grande, Brazil. The observed prevalence of HSV-2 was 15.6%. Among the 302 women studied, 158 had received assistance in BHU and 144 were treated at FURG. The prevalence of HSV-2 in these groups was 10.8% and 20.8%, respectively, RR 1.9 and p = 0.012. Knowledge about the Pap smear, and the presence of lesions showed no association with HSV-2 infection. Multivariate analysis showed that the variable that most influenced the risk of HSV-2 infection was the presence of HIV infection, with a relative risk of 1.9 and p = 0.04. Discussion: Genital ulcers are an important entry point for HIV, and condom use is an important strategy to reduce transmission of HIV and HSV-2. PMID:24037285

  1. PREVALENCE OF HERPES SIMPLEX VIRUS TYPE 2 AND RISK FACTORS ASSOCIATED WITH THIS INFECTION IN WOMEN IN SOUTHERN BRAZIL

    Directory of Open Access Journals (Sweden)

    Thais Duquia Moraes Caldeira

    2013-09-01

    Full Text Available SUMMARY The herpes simplex virus type 2 (HVS-2 is the most prevalent infection worldwide. It is a cofactor in the acquisition of human immunodeficiency virus (HIV and the persistence of human papillomavirus (HPV. This study evaluated the prevalence of HSV-2, using the polymerase chain reaction (PCR, and associated factors in patients treated at the Federal University of Rio Grande (FURG and Basic Health Units (BHU in Rio Grande, Brazil. The observed prevalence of HSV-2 was 15.6%. Among the 302 women studied, 158 had received assistance in BHU and 144 were treated at FURG. The prevalence of HSV-2 in these groups was 10.8% and 20.8%, respectively, RR 1.9 and p = 0.012. Knowledge about the Pap smear, and the presence of lesions showed no association with HSV-2 infection. Multivariate analysis showed that the variable that most influenced the risk of HSV-2 infection was the presence of HIV infection, with a relative risk of 1.9 and p = 0.04. Discussion: Genital ulcers are an important entry point for HIV, and condom use is an important strategy to reduce transmission of HIV and HSV-2.

  2. Detection of Mouse Mammary Tumour Virus in house mice

    DEFF Research Database (Denmark)

    Steffensen, Lise K; Leirs, Herwig; Heiberg, Ann-Charlotte

    The prevalence of human breast cancer (HBC) is affected by several parameters. For the past decades MMTV, Mouse Mammary Tumor Virus, known to cause breast cancer in mice, has been hypothesized to affect the frequency of hormone dependent HBC. Though conclusive evidence has not been produced, still...... and the fact that human cells can be infected in vitro strengthen the theory. Results from above mentioned studies indicate that the source of provirus is exogenous. The transmission mechanism is tentatively suggested to be by means of recombination of exogeneous and endogenous viruses obtained from...

  3. Detection of antibodies against Leishmania infantum in cats (Felis catus from the State of Pernambuco, Brazil

    Directory of Open Access Journals (Sweden)

    Rita de Cássia Nascimento Silva

    2014-01-01

    Full Text Available Introduction: Little information is available concerning infection by Leishmania infantum in cats. Therefore, the aim of this study was to perform a serological study in domestic cats. Methods: Serum samples (n=153 obtained from animals living in the Cities of Recife and Petrolina, State of Pernambuco, Brazil, were tested by ELISA/S7® (Biogene. Results: Anti-L. infantum antibodies were detected in 3.9% (6/153 of the cats. All seroreagent animals were from Petrolina. Conclusions: These results serve as an important alert, and future studies are needed to better understand the possible role of cats in the epidemiology of visceral leishmaniasis (VL in this area.

  4. Detection of Lawsonia intracellularis fecal shedding in dogs in Minas Gerais, Brazil

    Directory of Open Access Journals (Sweden)

    Rodrigo Otávio Silveira Silva

    2015-09-01

    Full Text Available Lawsonia intracellularisis an obligate intracellular bacterium that is responsible for proliferative enteropathy, an enteric disease endemic in swine and common in foals. However, few studies have investigated this disease in dogs, and there are no reports of dogs infected with L. intracellularisin Latin America. The aim of this study was to evaluate the fecal shedding of L. intracellularisin diarrheic and non-diarrheic dogs in Minas Gerais, Brazil. A total of 58 dogs, 18 apparently healthy and 40 diarrheic, were examined in this study. DNA extracted from feces was analyzed using a nested PCR reaction to detect L. intracellularis.Three out of 40 (7.5% diarrheic samples, all from 3-month-old puppies, were positive for L. intracellularis. These results highlight the need for additional studies to examine the role of this pathogen as a possible cause of enteric disease in dogs

  5. Molecular detection of Ehrlichia canis in dogs from the Pantanal of Mato Grosso State, Brazil.

    Science.gov (United States)

    Santos, Luana Gabriela Ferreira Dos; Melo, Andréia Lima Tomé; Moraes-Filho, Jonas; Witter, Rute; Labruna, Marcelo Bahia; Aguiar, Daniel Moura de

    2013-01-01

    The present study evaluated the presence of Ehrlichia DNA in the blood samples of 320 dogs from the urban and rural areas of the municipality of Poconé, Pantanal region, Mato Grosso state, by Polymerase Chain Reaction (PCR), targeting the ehrlichial dsb gene. Risk factors for infection in dogs were also evaluated. Forty-eight (15%, 95% CI: 11.4-19.5%) dogs were positive: 25 (15.6%, 95% CI: 10.4-22.2%) from the urban area and 23 (14.4%, 95% CI: 9.3-20.8%) from the rural area (P > 0.05). Partial DNA sequence obtained from PCR products of 18 samples from the urban area and 16 samples from the rural area were 100% identical to E. canis from Brazil and the USA. This study reports the first E. canis molecular detection in dogs from the northern Pantanal region.

  6. How Does Sampling Methodology Influence Molecular Detection and Isolation Success in Influenza A Virus Field Studies?

    Science.gov (United States)

    Latorre-Margalef, Neus; Avril, Alexis; Tolf, Conny; Olsen, Björn; Waldenström, Jonas

    2015-12-11

    Wild waterfowl are important reservoir hosts for influenza A virus (IAV) and a potential source of spillover infections in other hosts, including poultry and swine. The emergence of highly pathogenic avian influenza (HPAI) viruses, such as H5N1 and H5N8, and subsequent spread along migratory flyways prompted the initiation of several programs in Europe, North America, and Africa to monitor circulation of HPAI and low-pathogenicity precursor viruses (low-pathogenicity avian influenza [LPAI] viruses). Given the costs of maintaining such programs, it is essential to establish best practice for field methodologies to provide robust data for epidemiological interpretation. Here, we use long-term surveillance data from a single site to evaluate the influence of a number of parameters on virus detection and isolation of LPAI viruses. A total of 26,586 samples (oropharyngeal, fecal, and cloacal) collected from wild mallards were screened by real-time PCR, and positive samples were subjected to isolation in embryonated chicken eggs. The LPAI virus detection rate was influenced by the sample type: cloacal/fecal samples showed a consistently higher detection rate and lower cycle threshold (Ct) value than oropharyngeal samples. Molecular detection was more sensitive than isolation, and virus isolation success was proportional to the number of RNA copies in the sample. Interestingly, for a given Ct value, the isolation success was lower in samples from adult birds than in those from juveniles. Comparing the results of specific real-time reverse transcriptase (RRT)-PCRs and of isolation, it was clear that coinfections were common in the investigated birds. The effects of sample type and detection methods warrant some caution in interpretation of the surveillance data. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  7. Nucleic Acid, Antibody, and Virus Culture Methods to Detect Xenotropic MLV-Related Virus in Human Blood Samples

    Directory of Open Access Journals (Sweden)

    M. F. Kearney

    2011-01-01

    Full Text Available The MLV-related retrovirus, XMRV, was recently identified and reported to be associated with both prostate cancer and chronic fatigue syndrome. At the National Cancer Institute-Frederick, MD (NCI-Frederick, we developed highly sensitive methods to detect XMRV nucleic acids, antibodies, and replication competent virus. Analysis of XMRV-spiked samples and/or specimens from two pigtail macaques experimentally inoculated with 22Rv1 cell-derived XMRV confirmed the ability of the assays used to detect XMRV RNA and DNA, and culture isolatable virus when present, along with XMRV reactive antibody responses. Using these assays, we did not detect evidence of XMRV in blood samples ( or prostate specimens ( from two independent cohorts of patients with prostate cancer. Previous studies detected XMRV in prostate tissues. In the present study, we primarily investigated the levels of XMRV in blood plasma samples collected from patients with prostate cancer. These results demonstrate that while XMRV-related assays developed at the NCI-Frederick can readily measure XMRV nucleic acids, antibodies, and replication competent virus, no evidence of XMRV was found in the blood of patients with prostate cancer.

  8. Detecting genetic introgression: high levels of intersubspecific recombination found in Xylella fastidiosa in Brazil.

    Science.gov (United States)

    Nunney, Leonard; Yuan, Xiaoli; Bromley, Robin E; Stouthamer, Richard

    2012-07-01

    Documenting the role of novel mutation versus homologous recombination in bacterial evolution, and especially in the invasion of new hosts, is central to understanding the long-term dynamics of pathogenic bacteria. We used multilocus sequence typing (MLST) to study this issue in Xylella fastidiosa subsp. pauca from Brazil, a bacterium causing citrus variegated chlorosis (CVC) and coffee leaf scorch (CLS). All 55 citrus isolates typed (plus one coffee isolate) defined three similar sequence types (STs) dominated by ST11 (85%), while the remaining 22 coffee isolates defined two STs, mainly ST16 (74%). This low level of variation masked unusually large allelic differences (>1% divergence with no intermediates) at five loci (leuA, petC, malF, cysG, and holC). We developed an introgression test to detect whether these large differences were due to introgression via homologous recombination from another X. fastidiosa subspecies. Using additional sequencing around these loci, we established that the seven randomly chosen MLST targets contained seven regions of introgression totaling 2,172 bp of 4,161 bp (52%), only 409 bp (10%) of which were detected by other recombination tests. This high level of introgression suggests the hypothesis that X. fastidiosa subsp. pauca became pathogenic on citrus and coffee (crops cultivated in Brazil for several hundred years) only recently after it gained genetic variation via intersubspecific recombination, facilitating a switch from native hosts. A candidate donor is the subspecies infecting plum in the region since 1935 (possibly X. fastidiosa subsp. multiplex). This hypothesis predicts that nonrecombinant native X. fastidiosa subsp. pauca (not yet isolated) does not cause disease in citrus or coffee.

  9. Detecting Genetic Introgression: High Levels of Intersubspecific Recombination Found in Xylella fastidiosa in Brazil

    Science.gov (United States)

    Yuan, Xiaoli; Bromley, Robin E.; Stouthamer, Richard

    2012-01-01

    Documenting the role of novel mutation versus homologous recombination in bacterial evolution, and especially in the invasion of new hosts, is central to understanding the long-term dynamics of pathogenic bacteria. We used multilocus sequence typing (MLST) to study this issue in Xylella fastidiosa subsp. pauca from Brazil, a bacterium causing citrus variegated chlorosis (CVC) and coffee leaf scorch (CLS). All 55 citrus isolates typed (plus one coffee isolate) defined three similar sequence types (STs) dominated by ST11 (85%), while the remaining 22 coffee isolates defined two STs, mainly ST16 (74%). This low level of variation masked unusually large allelic differences (>1% divergence with no intermediates) at five loci (leuA, petC, malF, cysG, and holC). We developed an introgression test to detect whether these large differences were due to introgression via homologous recombination from another X. fastidiosa subspecies. Using additional sequencing around these loci, we established that the seven randomly chosen MLST targets contained seven regions of introgression totaling 2,172 bp of 4,161 bp (52%), only 409 bp (10%) of which were detected by other recombination tests. This high level of introgression suggests the hypothesis that X. fastidiosa subsp. pauca became pathogenic on citrus and coffee (crops cultivated in Brazil for several hundred years) only recently after it gained genetic variation via intersubspecific recombination, facilitating a switch from native hosts. A candidate donor is the subspecies infecting plum in the region since 1935 (possibly X. fastidiosa subsp. multiplex). This hypothesis predicts that nonrecombinant native X. fastidiosa subsp. pauca (not yet isolated) does not cause disease in citrus or coffee. PMID:22544234

  10. Detection of herpes viruses in the cerebrospinal fluid of adults with suspected viral meningitis in Malawi.

    Science.gov (United States)

    Benjamin, L A; Kelly, M; Cohen, D; Neuhann, F; Galbraith, S; Mallewa, M; Hopkins, M; Hart, I J; Guiver, M; Lalloo, D G; Heyderman, R S; Solomon, T

    2013-02-01

    We looked for herpes simplex virus types 1 and 2 (HSV-1 and HSV-2, respectively), varicella zoster virus (VZV), Epstein-Barr virus (EBV) and cytomegalovirus (CMV) DNA in Malawian adults with clinically suspected meningitis. We collected cerebrospinal fluid (CSF) from consecutive adults admitted with clinically suspected meningitis to Queen Elizabeth Central Hospital (QECH), Blantyre, Malawi, for a period of 3 months. Those with proven bacterial or fungal meningitis were excluded. Real-time polymerase chain reaction (PCR) was performed on the CSF for HSV-1 and HSV-2, VZV, EBV and CMV DNA. A total of 183 patients presented with clinically suspected meningitis. Of these, 59 (32 %) had proven meningitis (bacterial, tuberculous or cryptococcal), 39 (21 %) had normal CSF and 14 (8 %) had aseptic meningitis. For the latter group, a herpes virus was detected in 9 (64 %): 7 (50 %) had EBV and 2 (14 %) had CMV, all were human immunodeficiency virus (HIV)-positive. HSV-2 and VZV were not detected. Amongst those with a normal CSF, 8 (21 %) had a detectable herpes virus, of which 7 (88 %) were HIV-positive. The spectrum of causes of herpes viral meningitis in this African population is different to that in Western industrialised settings, with EBV being frequently detected in the CSF. The significance of this needs further investigation.

  11. Detection of brucellosis in water buffaloes for exportation in northern and northeastern of Brazil

    Directory of Open Access Journals (Sweden)

    José Diomedes Barbosa

    2016-11-01

    Full Text Available ABSTRACT. Barbosa J.D., Bomjardim H.dosA., Lima D.H.daS., Reis A.dosS.B., Barboza F.B., Albernaz T.T., Oliveira C.M.C., Fonseca A.H., Nicolino R.R. & da Silva J.B. Detection of brucellosis in water buffaloes for exportation in northern and northeastern of Brazil. [Detecção de brucelose em búfalos d’agua para exportação no norte e nordeste do Brasil}. Revista Brasileira de Medicina Veterinária, 38(supl. 3: 129-135, 2016. Laboratório de Doenças Parasitárias, Departamento de Epidemiologia e Saúde Pública, Instituto de Veterinária, Universidade Federal Rural de Rio de Janeiro, BR 465, Km 7 Seropédica, RJ 23890-000, Brazil. E-mail: adivaldofonseca@yahoo.com The prevalence of brucellosis in buffaloes was evaluated by the Buffered Acidified Plate Antigen (BAPA in 5.163 water buffaloes from Maranhão state, Pará state and Marajó Island, Brazil. The detection of buffaloes positive for brucellosis by BAPA was 7.37% in Marajó Island, 8.45% in Pará state and 29.86% in Maranhão state. The locations with the highest prevalences were Santa Cruz do Arari, in Marajó Island (12.50%; Ipixuna, in Pará state (30.25%; and Santa Inês, in Maranhão state (34.76%. After the confirmatory test (Complement Fixation Test, only 7 animals remained positive in Marajó Island, and 22 remained positive in the state of Pará. None of the 66 animals that reacted positively in the BAPA test in Maranhão reacted positively in the CF. The high prevalence of B. abortus that was observed in animals, especially in the state of Maranhão, is worrisome for the health system for the control and eradication of bovine brucellosis.

  12. Forecasting the new case detection rate of leprosy in four states of Brazil: A comparison of modelling approaches

    NARCIS (Netherlands)

    D.J. Blok (David); R.E. Crump (Ron E.); Sundaresh, R. (Ram); M. Ndeffo-Mbah (Martial); Galvani, A.P. (Alison P.); T.C. Porco (Travis C.); S.J. de Vlas (Sake); G.F. Medley (Graham F.); J.H. Richardus (Jan Hendrik)

    2017-01-01

    textabstractBackground Brazil has the second highest annual number of new leprosy cases. The aim of this study is to formally compare predictions of future new case detection rate (NCDR) trends and the annual probability of NCDR falling below 10/100,000 of four different modelling approaches in four

  13. Detection of mumps virus RNA in cerebrospinal fluid of patients with neuromyelitis optica.

    Science.gov (United States)

    Mori, Masahiro; Hosoya, Mitsuaki; Hiwasa, Takaki; Hayakawa, Sei; Uzawa, Akiyuki; Kuwabara, Satoshi

    2011-10-01

    Neuromyelitis optica (NMO) is an acute inflammatory disease that preferentially involves the optic nerves and spinal cord. Although many infectious agents, including mumps virus, are postulated to have a role in the pathogenesis of multiple sclerosis (MS), the relationship between NMO and infectious agents remains uncertain. To investigate the relationship between NMO and viruses that have special affinity for the central nervous system, we performed a nested polymerase chain reaction (PCR) to detect mumps virus or enterovirus RNA in cerebrospinal fluid samples from 13 patients with MS, 8 with NMO and 20 with other neurological diseases (ONDs). Nested PCR was positive for mumps virus in 2 (25%) of NMO patients, but in none of those with MS and ONDs. Moreover, nested PCR results became negative in the remission phase in the two PCR-positive NMO patients. Mumps virus may have some role in the pathogenesis of NMO.

  14. Detection of Serum Antibodies to Borna Disease Virus in Patients with Psychiatric Disorders

    Science.gov (United States)

    Rott, R.; Herzog, S.; Fleischer, B.; Winokur, A.; Amsterdam, J.; Dyson, W.; Koprowski, H.

    1985-05-01

    Borna disease virus causes a rare meningoencephalitis in horses and sheep and has been shown to produce behavioral effects in some species. The possibility that the Borna virus is associated with mental disorders in humans was evaluated by examining serum samples from 979 psychiatric patients and 200 normal volunteers for the presence of Borna virus-specific antibodies. Antibodies were detected by the indirect immunofluorescence focus assay. Antibodies to the virus were demonstrated in 16 of the patients but none of the normal volunteers. The patients with the positive serum samples were characterized by having histories of affective disorders, particularly of a cyclic nature. Further studies are needed to define the possible involvement of Borna virus in human psychiatric disturbances.

  15. Distribution and molecular detection of apple mosaic virus in apple ...

    African Journals Online (AJOL)

    ... 15 apple ApMV isolates were obtained. All of the amplicons were subjected to enzymatic digestion with restriction endonuclease enzymes and phylogenetic analysis were performed according to the digestion profiles. Keywords: Apple mosaic virus, coat protein gene, hazelnut. African Journal of Biotechnology, Vol 13(31) ...

  16. Detection of banana streak virus (BSV) Tamil Nadu isolate (India ...

    African Journals Online (AJOL)

    Yomi

    2012-10-09

    Oct 9, 2012 ... 2Department of Fruit Crops, Tamil Nadu Agricultural University, Coimbatore-641 003, Tamil Nadu, India. Accepted 5 September, 2012. Banana streak virus (BSV) is of quarantine significance since Musa is a vegetatively propagated crop. Diagnosis by symptomatology is unreliable because the symptoms ...

  17. Detection of bovine viral diarrhea virus antibodies in camels ...

    African Journals Online (AJOL)

    This study was carried out to determine the seroprevalence of bovine viral diarrhea virus (BVDV) antibodies in camels presented for slaughter at the Maiduguri abattoir using a BVDV specific indirect enzyme-linked-immunosorbent assay (ELISA). Ninety (90) serum samples collected from adult male and female camels were ...

  18. Detection of Infectious Bursal Disease Virus (IBDV) in naturally ...

    African Journals Online (AJOL)

    The Reverse Transcription - Polymerase Chain Reaction (RT-PCR) was used for the identification of Infectious bursal disease virus (IBDV). The technique was applied on bursa of Fabricius of infected chicken. Some of these bursae have been kept in the freezer for 16years under conditions of regular electric power ...

  19. Detection of avian nephritis virus and chicken astrovirus in Nigerian ...

    African Journals Online (AJOL)

    Avian nephritis virus (ANV) and chicken astrovirus (CAstV) are widely distributed in poultry flocks worldwide, causing growth retardation. However, these avian astroviruses have not been previously diagnosed in poultry species in Nigeria. Real-time reverse transcriptase-polymerase chain reaction (rRTPCR) and reverse ...

  20. Distribution and molecular detection of apple mosaic virus in apple ...

    African Journals Online (AJOL)

    SAM

    2014-07-30

    Jul 30, 2014 ... Apple mosaic virus (ApMV) is one of the most important diseases limiting the production of hazelnut and apple in Turkey and the objectives of this research were to determine the convenient and reliable method for RNA isolation and also to determine primer pair for real time polymerase chain reaction (RT-.

  1. Detection and distribution of sweetpotato feathery mottle virus in ...

    African Journals Online (AJOL)

    Owner

    Planck, 1968). Susceptibility is characterized by a total absence of reaction from the plant face to the large number of rapidly multiplying pathogens in all organs and tissues of attacked plants (Semal et al., 1994). A dosage of virus concentration indicates an important concentration of the pathogen in the tissues of the.

  2. Analytical methods for virus detection in water and food

    NARCIS (Netherlands)

    Bosch, A.; Sanchez, G.; Abbaszadegan, M.; Carducci, A.; Guix, S.; Guyader, Le F.S.; Netshikweta, R.; Pintó, R.M.; Poel, van der W.H.M.; Rutjes, S.; Sano, D.; Taylor, M.D.; Zijl, Van W.B.; Rodriguez-Lázaro, D.; Kovac, K.; Sellwood, J.

    2011-01-01

    Potential ways to address the issues that relate to the techniques for analyzing food and environmental samples for the presence of enteric viruses are discussed. It is not the authors’ remit to produce or recommend standard or reference methods but to address specific issues in the analytical

  3. Simultaneous detection of respiratory syncytial virus types A and B ...

    African Journals Online (AJOL)

    Multiplex RT-PCR has a significant advantage in that it permits the simultaneous amplification of several viruses in a single reaction facilitating cost-effective diagnosis and perhaps improved clinical management. Objectives: In this study, our aim was to determine the prevalence of influenza A and B, and RSV types A and B ...

  4. Serological detection of viruses infecting tomato and pepper in ...

    African Journals Online (AJOL)

    Mixed viral infections were few, PVY + PVMV occurring only in one tomato leaf sample while PVMV + CMV occurred on three pepper leaf samples. The control of aphid vectors that transmit these viruses and good sanitary practices against soil borne ToMV would minimize disease incidences and subsequent yield loss.

  5. serological detection of seed borne viruses in cowpea regenerated ...

    African Journals Online (AJOL)

    Administrator

    mosaic virus genus Potyvirus, which is of great economic importance in germplasm conservation and exchange between ... onto hormone free medium composed of 4.43 g MS basal salt, 30 g sucrose and 8 g of agar. ... la conservation du germoplasme et l'échange entre les pays eu égard aux pertes qu'il entraîne.

  6. Detection and elimination of sweetpotato viruses | Rukarwa | African ...

    African Journals Online (AJOL)

    Based on shoot survival and effectiveness of virus elimination, the best results were obtained by exposing plantlets to daily temperature regime of 32 oC for 8 hr of darkness and 36 oC for 16 hr of light for four weeks. Meristem-tip culture combined with thermotherapy allowed elimination of SPFMV and SPMMV in 77% of ...

  7. Rapid detection of Hendra virus antibodies: an integrated device with nanoparticle assay and chaotic micromixing.

    Science.gov (United States)

    Petkovic, K; Metcalfe, G; Chen, H; Gao, Y; Best, M; Lester, D; Zhu, Y

    2016-12-20

    Current diagnosis of infectious diseases such as Hendra virus (HeV) relies mostly on laboratory-based tests. There is an urgent demand for rapid diagnosis technology to detect and identify these diseases in humans and animals so that disease spread can be controlled. In this study, an integrated lab-on-a-chip device using a magnetic nanoparticle immunoassay is developed. The key features of the device are the chaotic fluid mixing, achieved by magnetically driven motion of nanoparticles with the optimal mixing protocol developed using chaotic transport theory, and the automatic liquid handling system for loading reagents and samples. The device has been demonstrated to detect Hendra virus antibodies in dilute horse serum samples within a short time of 15 minutes and the limit of detection is about 0.48 ng ml -1 . The device platform can potentially be used for field detection of viruses and other biological and chemical substances.

  8. Development and evaluation of a Luminex multiplex serology assay to detect antibodies to bovine herpes virus 1, parainfluenza 3 virus, bovine viral diarrhoea virus, and bovine respiratory syncytial virus, with comparison to existing ELISA detection methods.

    Science.gov (United States)

    Anderson, Steve; Wakeley, Phil; Wibberley, Guy; Webster, Kath; Sawyer, Jason

    2011-03-07

    Detection of circulating antibodies to bovine herpes virus 1 (BHV-1), parainfluenza 3 virus (PI3V), bovine viral diarrhoea virus (BVDV) and bovine respiratory syncytial virus (BRSV) using ELISA is widely used for veterinary diagnostics and surveillance. In this paper, the potential of a multiplex serology test based on Luminex technology, where all antibodies are simultaneously detected in a single assay was investigated. The performance of "in-house" separate ELISAs which use relatively crude lysates of cultured virus as capture antigens, was compared to the multiplex assay where the same antigens were covalently bound to the fluorescent beads used in the Luminex platform. A panel of field serum samples was tested by the multiplex assay in parallel with the separate routine ELISAs to provide a comparison between tests. The BHV-1 and PI3V components of the multiplex test showed similar sensitivities and specificities to the separate "in-house" ELISAs. The performance of the BVDV and BRSV components was less successful and was attributed to relatively low signal strength for these antigens, leading to higher assay variability and a reduced ability to distinguish positive and negative samples compared to the "in-house" ELISAs. The results illustrated that antigens commonly used successfully in ELISAs cannot always be transferred for use in alternative assay systems. The use of recombinant BVDV E2 protein was investigated and was shown to lead to an appreciable increase in signal strength compared to the use of crude BVDV antigen in the Luminex system. Crown Copyright © 2011. Published by Elsevier B.V. All rights reserved.

  9. Molecular detection of Trypanosoma evansi (Kinetoplastida: Trypanosomatidae in procyonids (Carnivora: Procyonidae in Eastern Amazon, Brazil

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    Paulo Cesar Magalhães-Matos

    2016-04-01

    Full Text Available ABSTRACT: The present study aimed to diagnose the natural infection of captive and free-living procyonids with Trypanosoma evansi in the states of Amapá and Pará, Brazil. From February 2012 to August 2013, whole blood samples and blood smears were obtained from 45 free-living procyonids and from nine procyonids kept in captivity in wild life refuges and zoobotanical parks in the states of Amapá and Pará. Whole blood samples were collected and kept at -20ºC for the detection of T. evansi DNA by PCR using the RoTat 1.2 forward and RoTat 1.2 reverse primers. In addition, the blood smears were processed and examined for the presence of trypomastigote forms of T. evansi. T. evansi DNA was detected in 18.52% (10/54 of the procyonids, namely, in captive crab-eating raccoons and captive and free-living coatis in Pará State. No trypomastigote forms were observed in the blood smears. DNA from T. evansi was detected in P. cancrivorus and N. nasua in Pará State, being this the first such report in P. cancrivorus.

  10. A liquid phase blocking ELISA for the detection of antibodies against infectious bronchitis virus

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    Cardoso T.C.

    1999-01-01

    Full Text Available A liquid phase blocking ELISA (LPB-ELISA was developed for the detection and measurement of antibodies against infectious bronchitis virus (IBV. The purified and nonpurified virus used as antigen, the capture and detector antibodies, and the chicken hyperimmune sera were prepared and standardized for this purpose. A total of 156 sera from vaccinated and 100 from specific pathogen-free chickens with no recorded contact with the virus were tested. The respective serum titers obtained in the serum neutralization test (SNT were compared with those obtained in the LPB-ELISA. There was a high correlation (r2 = 0.8926 between the two tests. The LPB-ELISA represents a single test suitable for the rapid detection of antibodies against bronchitis virus in chicken sera, with good sensitivity (88%, specificity (100% and agreement (95.31%.

  11. Quantitative detection of Merkel cell virus in human tissues and possible mode of transmission.

    Science.gov (United States)

    Loyo, Myriam; Guerrero-Preston, Rafael; Brait, Mariana; Hoque, Mohmammad O; Chuang, Alice; Kim, Myoung S; Sharma, Rajni; Liégeois, Nanette J; Koch, Wayne M; Califano, Joseph A; Westra, William H; Sidransky, David

    2010-06-15

    Merkel Cell Virus (MCV) is a newly discovered polyomavirus, recently found in a rare skin cancer, Merkel cell carcinoma (MCC). However, MCV has also been detected in some normal tissue samples. We tested and compared the relative quantity of the MCV in a set of diverse human tissue samples with the MCC samples. The levels of MCV in MCCs were over 60 times higher than the highest values in all other tissues. Low quantities of MCV were detected in diverse tissue samples independently of malignant or benign histologic status. Higher levels of the virus were found in the upper aerodigestive tract, digestive system, and saliva compared to the lung and genitourinary system samples. These results confirm that MCV is widespread in the human body and suggest a possible fecal-oral transmission route similar to the Hepatitis A virus. Despite widespread presence of the virus, it appears that only neuroendocrine skin cells are susceptible to transformation by MCV.

  12. Natural transovarial transmission of dengue virus 4 in Aedes aegypti from Cuiabá, State of Mato Grosso, Brazil

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    Lucinéia Claudia de Toni Aquino da Cruz

    2015-02-01

    Full Text Available INTRODUCTION: Dengue is the most prevalent arboviral disease in tropical areas. In Mato Grosso, outbreaks are reported every year, but studies on dengue in this state are scarce. METHODS: Natural transovarial infection of Aedes aegypti by a flavivirus was investigated in the Jardim Industriário neighborhood of Cuiabá, Mato Grosso. Eggs were collected with ovitraps during the dry, intermediate, and rainy seasons of 2012. After the eggs hatched and the larvae developed to adulthood, mosquitoes (n = 758 were identified and allocated to pools of 1-10 specimens according to the collection location, sex, and climatic period. After RNA extraction, multiplex semi-nested RT-PCR was performed to detect the four dengue virus (DENV serotypes, yellow fever virus, West Nile virus and Saint Louis encephalitis virus. RESULTS: DENV-4 was the only flavivirus detected, and it was found in 8/50 pools (16.0%. Three of the positive pools contained females, and five contained males. Their nucleotide sequences presented 96-100% similarity with DENV-4 genotype II strains from Manaus, Amazonas. The minimum infection rate was 10.5 per 1000 specimens, and the maximum likelihood estimator of the infection rate was 11.6 (95% confidence interval: 4.8; 23.3. CONCLUSIONS: This study provides the first evidence of natural transovarial infection by DENV-4 in Ae. Aegypti in Mato Grosso, suggesting that this type of infection might serve as a mechanism of virus maintenance during interepidemic periods in Cuiabá, a city where dengue epidemics are reported every year. These results emphasize the need for efficient vector population control measures to prevent arbovirus outbreaks in the state.

  13. Development of a blocking latex agglutination test for the detection of antibodies to chicken anemia virus.

    Science.gov (United States)

    Trinh, Dai Quang; Ogawa, Haruko; Bui, Vuong Nghia; Nguyen, Tham Thi Hong; Gronsang, Dulyatad; Baatartsogt, Tugsbaatar; Kizito, Mugimba Kahoza; AboElkhair, Mohammed; Yamaguchi, Shigeo; Nguyen, Viet Khong; Imai, Kunitoshi

    2015-09-01

    A blocking latex agglutination test (b-LAT) developed in this study was evaluated for the detection of antibodies against chicken anemia virus (CAV) in chickens. Polystyrene latex beads were coupled with a neutralizing monoclonal antibody (mAb) to CAV (mAb-beads). When mAb-beads were mixed with antigens prepared from the lysate of MDCC-MSB1 cells infected with CAV, agglutination occurred. A short pre-incubation of CAV antigens with CAV-specific antiserum inhibited the agglutination of mAb-beads. The test results were obtained within 5min. The specificity of b-LAT was evaluated using sera from specific pathogen-free chickens and sera containing antibodies to avian influenza virus, Newcastle disease virus, infectious bursal disease virus, and Marek's disease virus; nonspecific agglutination and cross-reactivity with antibodies to unrelated viruses were not observed. The examination of 94 serum samples collected from commercial breeder chickens of various ages (17-63 weeks) revealed good agreement (93.6%, Kappa value=0.82) between b-LAT and a virus neutralization test, known to be most sensitive and specific in the detection of antibodies to CAV. These results indicate that b-LAT, a simple and rapid test, is a useful and reliable tool in CAV serology. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. Detection of infectious laryngotracheitis virus by real-time PCR in naturally and experimentally infected chickens.

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    Yan Zhao

    Full Text Available Infectious laryngotracheitis (ILT is an acute, highly contagious upper-respiratory infectious disease of chickens. In this study, a real-time PCR method was developed for fast and accurate detection and quantitation of ILTV DNA of chickens experimentally infected with ILTV strain LJS09 and naturally infected chickens. The detection lower limit of the assay was 10 copies of DNA. There were no cross reactions with the DNA and RNA of infectious bursal disease virus, chicken anemia virus, reticuloendotheliosis virus, avian reovirus, Newcastle disease virus, and Marek's disease virus. The real-time PCR was reproducible as the coefficients of variation of reproducibility of the intra-assay and the inter-assay were less than 2%. The real-time PCR was used to detect the levels of the ILTV DNA in the tissues of specific pathogen free (SPF chickens infected with ILTV at different times post infection. ILTV DNA was detected by real-time PCR in the heart, liver, spleen, lung, kidney, larynx, tongue, thymus, glandular stomach, duodenum, pancreatic gland, small intestine, large intestine, cecum, cecal tonsil, bursa of Fabricius, and brain of chickens in the infection group and the contact-exposure group. The sensitivity, specificity, and reproducibility of the ILTV real-time PCR assay revealed its suitability for detection and quantitation of ILTV in the samples from clinically and experimentally ILTV infected chickens.

  15. Optical biosensor system for the quick and reliable detection of virus infections: VIROSENS

    Science.gov (United States)

    Proll, Günther; Hartjes, Anja; Sinclair, Alexander; Markovic, Goran; Pröll, Florian; Patel, Pranav; Niedrig, Matthias

    2014-10-01

    Viral infections are of special threat because they can induce severe courses of disease but only few medical treatments are available. Because of socio-economic and climate changes, increased worldwide mobility and population growth, the risk of newly occurring and quickly spreading viral pathogens has increased. A diagnosis of these diseases at an early stage is essential for a quick risk assessment and a proper health management as well as patient's treatment in an optimal way. Currently, the diagnosis of such diseases is based on time consuming and costly detection methods that can only be performed by specially trained personnel in laboratories at specific security levels. Aim of the project VIROSENS is the development of a biosensor platform that can specifically detect virus particles as well as virus-specific antibodies out of biological matrices like blood, serum, plasma and other body fluids. For this purpose, a disposable cartridge for such antibody- and virus-arrays is designed and developed within the project. The optical detection of viruses is performed with a portable device that will be benchmarked and evaluated concerning currently used standard detection methods in terms of its analytical performance. Within this project, a novel combination of serological tests and direct detection of virus particles will be developed, which will provide faster and more reliable results than presently available and used test systems.

  16. Comparison of three multiplex gastrointestinal platforms for the detection of gastroenteritis viruses.

    Science.gov (United States)

    Chhabra, Preeti; Gregoricus, Nicole; Weinberg, Geoffrey A; Halasa, Natasha; Chappell, James; Hassan, Ferdaus; Selvarangan, Rangaraj; Mijatovic-Rustempasic, Slavica; Ward, M Leanne; Bowen, Michael; Payne, Daniel C; Vinjé, Jan

    2017-10-01

    Viruses are major etiological agents of childhood gastroenteritis. In recent years, several molecular platforms for the detection of viral enteric pathogens have become available. We evaluated the performance of three multiplex platforms including Biofire's Gastrointestinal Panel (FilmArray), Luminex xTAG ® Gastrointestinal Pathogen Panel (GPP), and the TaqMan Array Card (TAC) for the detection of five gastroenteritis viruses using a coded panel of 300 archived stool samples. The FilmArray detected a virus in 199 (96.1%) and the TAC in 172 (83.1%) of the 207 samples (187 samples positive for a single virus and 20 samples positive for more than one virus) whereas the GPP detected a virus in 100 (78.7%) of the 127 (97 positive for one virus and three positive for more than one virus) samples. Overall the clinical accuracy was highest for the FilmArray (98%) followed by TAC (97.2%) and GPP (96.9%). The sensitivity of the FilmArray, GPP and TAC platforms was highest for rotavirus (100%, 95.8%, and 89.6%, respectively) and lowest for adenovirus type 40/41 (97.4%, 57.9% and 68.4%). The specificity of the three platforms ranged from 95.6% (rotavirus) to 99.6% (norovirus/sapovirus) for the FilmArray, 99.6% (norovirus) to 100% (rotavirus/adenovirus) for GPP, and 98.9% (astrovirus) to 100% (rotavirus/sapovirus) for TAC. The FilmArray demonstrated the best analytical performance followed by TAC. In recent years, the availability of multi-enteric molecular testing platforms has increased significantly and our data highlight the strengths and weaknesses of these platforms. Published by Elsevier B.V.

  17. Comparative evaluation of coagglutination and latex agglutination test (Rotalex kit) for detection of rota virus.

    OpenAIRE

    Mathur M; Bhave G

    1993-01-01

    Coagglutination test was compared with commercially available latex agglutination test (Rotalex kit) for detection of rota virus in faecal samples from clinically suspected cases of viral gastroenteritis. Out of 80 test samples 16 (20%) and 20 (25.3%) were positive for rota virus antigen by Rotalex kit and coagglutination test respectively. All the 40 controls were negative for viral antigen by Rotalex kit and only one gave positive result by coagglutination test. Coagglutinatio...

  18. Comparative evaluation of coagglutination and latex agglutination test (Rotalex kit for detection of rota virus.

    Directory of Open Access Journals (Sweden)

    Mathur M

    1993-07-01

    Full Text Available Coagglutination test was compared with commercially available latex agglutination test (Rotalex kit for detection of rota virus in faecal samples from clinically suspected cases of viral gastroenteritis. Out of 80 test samples 16 (20% and 20 (25.3% were positive for rota virus antigen by Rotalex kit and coagglutination test respectively. All the 40 controls were negative for viral antigen by Rotalex kit and only one gave positive result by coagglutination test. Coagglutination test was found to be economical, sensitive and specific for screening and rapid diagnosis of Rota virus diarrhoea.

  19. Simultaneous detection of papaya ringspot virus, papaya leaf distortion mosaic virus, and papaya mosaic virus by multiplex real-time reverse transcription PCR.

    Science.gov (United States)

    Huo, P; Shen, W T; Yan, P; Tuo, D C; Li, X Y; Zhou, P

    2015-12-01

    Both the single infection of papaya ringspot virus (PRSV), papaya leaf distortion mosaic virus (PLDMV) or papaya mosaic virus (PapMV) and double infection of PRSV and PLDMV or PapMV which cause indistinguishable symptoms, threaten the papaya industry in Hainan Island, China. In this study, a multiplex real-time reverse transcription PCR (RT-PCR) was developed to detect simultaneously the three viruses based on their distinctive melting temperatures (Tms): 81.0±0.8°C for PRSV, 84.7±0.6°C for PLDMV, and 88.7±0.4°C for PapMV. The multiplex real-time RT-PCR method was specific and sensitive in detecting the three viruses, with a detection limit of 1.0×10(1), 1.0×10(2), and 1.0×10(2) copies for PRSV, PLDMV, and PapMV, respectively. Indeed, the reaction was 100 times more sensitive than the multiplex RT-PCR for PRSV, and 10 times more sensitive than multiplex RT-PCR for PLDMV. Field application of the multiplex real-time RT-PCR demonstrated that some non-symptomatic samples were positive for PLDMV by multiplex real-time RT-PCR but negative by multiplex RT-PCR, whereas some samples were positive for both PRSV and PLDMV by multiplex real-time RT-PCR assay but only positive for PLDMV by multiplex RT-PCR. Therefore, this multiplex real-time RT-PCR assay provides a more rapid, sensitive and reliable method for simultaneous detection of PRSV, PLDMV, PapMV and their mixed infections in papaya.

  20. Highly Specific Detection of Five Exotic Quarantine Plant Viruses using RT-PCR

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    Hoseong Choi

    2013-03-01

    Full Text Available To detect five plant viruses (Beet black scorch virus, Beet necrotic yellow vein virus, Eggplant mottled dwarf virus, Pelargonium zonate spot virus, and Rice yellow mottle virus for quarantine purposes, we designed 15 RT-PCR primer sets. Primer design was based on the nucleotide sequence of the coat protein gene, which is highly conserved within species. All but one primer set successfully amplified the targets, and gradient PCRs indicated that the optimal temperature for the 14 useful primer sets was 51.9°C. Some primer sets worked well regardless of annealing temperature while others required a very specific annealing temperature. A primer specificity test using plant total RNAs and cDNAs of other plant virus-infected samples demonstrated that the designed primer sets were highly specific and generated reproducible results. The newly developed RT-PCR primer sets would be useful for quarantine inspections aimed at preventing the entry of exotic plant viruses into Korea.

  1. Detection and characterization of Aichi virus 1 in pediatric patients with diarrhea in Thailand.

    Science.gov (United States)

    Chuchaona, Watchaporn; Khamrin, Pattara; Yodmeeklin, Arpaporn; Kumthip, Kattareeya; Saikruang, Wilaiporn; Thongprachum, Aksara; Okitsu, Shoko; Ushijima, Hiroshi; Maneekarn, Niwat

    2017-02-01

    Kobuvirus is a newly discovered virus that belongs to the Kobuvirus genus in Picornaviridae family, which comprised of three species including Aichivirus A, Aichivirus B, and Aichivirus C. The kobuvirus isolated from human has been classified as Aichi virus 1 and belongs to Aichivirus A species. The present study aimed to assess the epidemiology and to perform molecular characterization of Aichi virus 1 in children admitted to hospitals with acute gastroenteritis in Chiang Mai, Thailand. A total of 923 fecal specimens collected from January, 2011 to December, 2013 were screened for the presence of Aichi virus 1 by RT semi-nested PCR. Out of 923 fecal specimens tested, Aichi virus 1 was detected with the prevalence of 2.6% (24/923). Of these, 0.3% (3/923) was genotype A and 2.3% (21/923) were genotype B. It is interesting to note that the genotype A showed the nucleotide sequence closely related to the Aichi virus reference strain isolated from sewage in Tunisia, while genotype B was most closely related to other human Aichi virus B reference strains. The results suggest that Aichi virus 1 of both genotypes A and B are circulating in pediatric patients in Thailand. J. Med. Virol. 89:234-238, 2017. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  2. Quantitative detection of four pome fruit viruses in apple trees throughout the year

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    Lucie WINKOWSKA

    2016-07-01

    Full Text Available A one-step real-time RT-PCR assay (RT-qPCR with melting curve analysis, using the green fluorescence dye SYBR Green I, was developed to detect and quantify RNA targets from Apple mosaic virus (ApMV, Apple stem grooving virus (ASGV, Apple stem pitting virus (ASPV and Apple chlorotic leaf spot virus (ACLSV in infected apple trees. Single PCR products of 87 bp (ApMV, 70 bp (ASGV, 104 bp (ASPV and 148 bp (ACLSV were obtained, and melting curve analyses revealed distinct melting temperature peaks for each virus. A dilution series using in vitro synthesized transcripts containing the target sequences as standards yielded a reproducible quantitative assay, with a wide dynamic range of detection and low coefficients of variance. The content of selected viruses in apple plant tissues was stable throughout the year, and their accumulation did not significantly change between different plant tissues. The only minor exceptions were for ApMV and ACLSV, in which noticeable differences in their concentrations in various biological material were observed within the year. This divergence did not influence their year-round detectability. This one-step RT-qPCR assay is a valuable tool for year-round diagnostics, and molecular studies of the biology of ApMV, ASGV, ASPV and ACLSV.

  3. Simultaneous detection of three lily viruses using Triplex IC-RT-PCR.

    Science.gov (United States)

    Zhang, Yubao; Wang, Yajun; Xie, Zhongkui; Yang, Guo; Guo, Zhihong; Wang, Le

    2017-11-01

    Viruses commonly infecting lily (Lilium spp.) include: Lily symptomless virus (LSV), Cucumber mosaic virus (CMV) and Lily mottle virus (LMoV). These viruses usually co-infect lilies causing severe economic losses in terms of quantity and quality of flower and bulb production around the world. Reliable and precise detection systems need to be developed for virus identification. We describe the development of a triplex immunocapture (IC) reverse transcription (RT) polymerase chain reaction (PCR) assay for the simultaneous detection of LSV, CMV and LMoV. The triplex IC-RT-PCR was compared with a quadruplex RT-PCR assay. Relative to the quadruplex RT-PCR, the specificity of the triplex IC-RT-PCR system for LSV, CMV and LMoV was 100% for field samples. The sensitivity of the triplex IC-RT-PCR system was 99.4%, 81.4% and 98.7% for LSV, CMV and LMoV, respectively. Agreement (κ) between the results obtained from the two tests was 0.968, 0.844 and 0.984 for LSV, CMV and LMoV, respectively. This is the first report of the simultaneous detection of LSV, CMV and LMoV in a triplex IC-RT-PCR assay. In particular we believe this convenient and reliable triplex IC-RT-PCR method could be used routinely for large-scale field surveys or crop health monitoring of lily. Copyright © 2017. Published by Elsevier B.V.

  4. Acanthamoeba polyphaga mimivirus stability in environmental and clinical substrates: implications for virus detection and isolation.

    Science.gov (United States)

    Dornas, Fábio P; Silva, Lorena C F; de Almeida, Gabriel M; Campos, Rafael K; Boratto, Paulo V M; Franco-Luiz, Ana P M; La Scola, Bernard; Ferreira, Paulo C P; Kroon, Erna G; Abrahão, Jônatas S

    2014-01-01

    Viruses are extremely diverse and abundant and are present in countless environments. Giant viruses of the Megavirales order have emerged as a fascinating research topic for virologists around the world. As evidence of their ubiquity and ecological impact, mimiviruses have been found in multiple environmental samples. However, isolation of these viruses from environmental samples is inefficient, mainly due to methodological limitations and lack of information regarding the interactions between viruses and substrates. In this work, we demonstrate the long-lasting stability of mimivirus in environmental (freshwater and saline water) and hospital (ventilator plastic device tube) substrates, showing the detection of infectious particles after more than 9 months. In addition, an enrichment protocol was implemented that remarkably increased mimivirus detection from all tested substrates, including field tests. Moreover, biological, morphological and genetic tests revealed that the enrichment protocol maintained mimivirus particle integrity. In conclusion, our work demonstrated the stability of APMV in samples of environmental and health interest and proposed a reliable and easy protocol to improve giant virus isolation. The data presented here can guide future giant virus detection and isolation studies.

  5. Acanthamoeba polyphaga mimivirus stability in environmental and clinical substrates: implications for virus detection and isolation.

    Directory of Open Access Journals (Sweden)

    Fábio P Dornas

    Full Text Available Viruses are extremely diverse and abundant and are present in countless environments. Giant viruses of the Megavirales order have emerged as a fascinating research topic for virologists around the world. As evidence of their ubiquity and ecological impact, mimiviruses have been found in multiple environmental samples. However, isolation of these viruses from environmental samples is inefficient, mainly due to methodological limitations and lack of information regarding the interactions between viruses and substrates. In this work, we demonstrate the long-lasting stability of mimivirus in environmental (freshwater and saline water and hospital (ventilator plastic device tube substrates, showing the detection of infectious particles after more than 9 months. In addition, an enrichment protocol was implemented that remarkably increased mimivirus detection from all tested substrates, including field tests. Moreover, biological, morphological and genetic tests revealed that the enrichment protocol maintained mimivirus particle integrity. In conclusion, our work demonstrated the stability of APMV in samples of environmental and health interest and proposed a reliable and easy protocol to improve giant virus isolation. The data presented here can guide future giant virus detection and isolation studies.

  6. Molecular surveillance of the Newcastle disease virus in domestic and wild birds on the North Eastern Coast and Amazon biome of Brazil

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    LM Thomazelli

    2012-03-01

    Full Text Available Brazil is one of the world's largest countries with a rich diversity of wildlife, including resident and migratory wild birds, which may be natural reservoirs of the Newcastle disease virus (NDV. Because Brazil is a major global exporter of chicken meat, the emergence of such a disease may have a huge negative impact not only on the economy due to trade restrictions and embargoes, but also on the quality of life of the population. Samples were collected from 1,022 asymptomatic domestic and wild birds from the Brazilian coast and the Amazon region using tracheal/cloacal swabs and tested by RT-qPCR. The results showed 7 (0.7% birds were positive for NDV. The positive samples were then isolated in embryonated chicken eggs and their matrix protein genes were partially sequenced, revealing a low-pathogenicity NDV. This study confirms the maintenance of the velogenic-NDV free status of Brazil.

  7. One-step RT-PCR for detection of Zika virus.

    Science.gov (United States)

    Faye, Oumar; Faye, Ousmane; Dupressoir, Anne; Weidmann, Manfred; Ndiaye, Mady; Alpha Sall, Amadou

    2008-09-01

    Zika virus (ZIKV) is an emerging mosquito-borne flavivirus circulating in Asia and Africa. Human infection induces an influenza-like syndrome that is associated with retro-orbital pain, oedema, lymphadenopathy, or diarrhea. Diagnosis of Zika fever requires virus isolation and serology, which are time consuming or cross-reactive. To develop a one-step RT-PCR assay to detect ZIKV in human serum. An assay targeting the envelope protein coding region was designed and evaluated for its specificity, detection limit, repeatability, and capacity to detect ZIKV isolates collected over a 40-year period from various African countries and hosts. The assay's detection limit and repeatability were respectively 7.7pfu/reaction and 100% in serum and L-15 medium; none of 19 other flaviviruses tested were detected. The assay is rapid, sensitive, and specific to detect ZIKV in cell culture or serum, but needs to be validated for diagnosis using clinical samples.

  8. Performance Evaluation of Allplex Respiratory Panels 1, 2, and 3 for Detection of Respiratory Viruses and Influenza A Virus Subtypes

    OpenAIRE

    Huh, Hee Jae; Kim, Ji-Youn; Kwon, Hyeon Jeong; Yun, Sun Ae; Lee, Myoung-Keun; Lee, Nam Yong; Kim, Jong-Won; Ki, Chang-Seok

    2017-01-01

    The Allplex respiratory panels 1, 2, and 3 (Allplex) comprise a one-step real-time reverse transcription-PCR assay for the detection of respiratory viruses (RVs) and influenza A subtypes based on multiple detection temperature (MuDT) technology. The performance of the Allplex assay was compared with those of the AdvanSure RV real-time PCR kit (AdvanSure) and the PowerChek pandemic H1N1/H3N2/H5N1 real-time PCR kit (PowerChek) using 417 clinical respiratory specimens. In comparison with the Adv...

  9. Basal core promoter and precore mutations among hepatitis B virus circulating in Brazil and its association with severe forms of hepatic diseases

    OpenAIRE

    Chachá, Silvana Gama Florencio; Gomes-Gouvêa, Michele Soares; Malta, Fernanda de Mello; Ferreira, Sandro da Costa; Villanova, Márcia Guimarães; Souza, Fernanda Fernandes; Teixeira, Andreza Correa; Passos, Afonso Dinis da Costa; Pinho, João Renato Rebello; Martinelli, Ana de Lourdes Candolo

    2017-01-01

    BACKGROUND In Brazil, few studies have investigated the prevalence of infection with the precore (PC) and basal core promoter (BCP) mutants of the hepatitis B virus (HBV). OBJECTIVES This study aimed to analyse the frequency of PC and BCP mutations among patients infected with HBV and to evaluate the association between the variants and advanced hepatic disease. METHODS A total of 161 patients infected with HBV were studied. To identify PC and BCP mutations, a 501-bp fragment of HBV DNA ...

  10. Detection of Bovine Leukemia Virus in Brains of Cattle with a Neurological Syndrome: Pathological and Molecular Studies

    Directory of Open Access Journals (Sweden)

    Rubens Henrique Ramos D’Angelino

    2013-01-01

    Full Text Available Bovine leukemia virus (BLV was investigated in the central nervous system (CNS of cattle with neurological syndrome. A total of 269 CNS samples were submitted to nested-PCR (BLV env gene gp51, and the viral genotypes were identified. The nested-PCR was positive in 4.8% (13/269 CNS samples, with 2.7% (2/74 presenting at histological examination lesions of nonpurulent meningoencephalitis (NPME, whereas 5.6% (11/195 not presenting NPME (P>0.05. No samples presented lymphosarcoma. The PCR products (437 bp were sequenced and submitted to phylogenetic analysis by neighbor-joining and maximum composite likelihood methods, and genotypes 1, 5, and 6 were detected, corroborating other South American studies. The genotype 6 barely described in Brazil and Argentina was more frequently detected in this study. The identity matrices showed maximum similarity (100% among some samples of this study and one from Argentina (FJ808582, recovered from GenBank. There was no association among the genotypes and NPME lesions.

  11. Detection of Bovine Leukemia Virus in Brains of Cattle with a Neurological Syndrome: Pathological and Molecular Studies

    Science.gov (United States)

    D'Angelino, Rubens Henrique Ramos; Pituco, Edviges Maristela; Villalobos, Eliana Monteforte Cassaro; Harakava, Ricardo; Gregori, Fábio

    2013-01-01

    Bovine leukemia virus (BLV) was investigated in the central nervous system (CNS) of cattle with neurological syndrome. A total of 269 CNS samples were submitted to nested-PCR (BLV env gene gp51), and the viral genotypes were identified. The nested-PCR was positive in 4.8% (13/269) CNS samples, with 2.7% (2/74) presenting at histological examination lesions of nonpurulent meningoencephalitis (NPME), whereas 5.6% (11/195) not presenting NPME (P > 0.05). No samples presented lymphosarcoma. The PCR products (437 bp) were sequenced and submitted to phylogenetic analysis by neighbor-joining and maximum composite likelihood methods, and genotypes 1, 5, and 6 were detected, corroborating other South American studies. The genotype 6 barely described in Brazil and Argentina was more frequently detected in this study. The identity matrices showed maximum similarity (100%) among some samples of this study and one from Argentina (FJ808582), recovered from GenBank. There was no association among the genotypes and NPME lesions. PMID:23710448

  12. Antibodies Against Henipa-Like Viruses in Brazilian Bats.

    Science.gov (United States)

    de Araujo, Jansen; Lo, Michael K; Tamin, Azaibi; Ometto, Tatiana L; Thomazelli, Luciano M; Nardi, Marcello S; Hurtado, Renata F; Nava, Alessandra; Spiropoulou, Christina F; Rota, Paul A; Durigon, Edison L

    2017-04-01

    Bats are reservoir hosts for many paramyxoviruses, some of which cause human and zoonotic diseases of public health importance. We developed a Nipah virus nucleoprotein enzyme-linked immunosorbent assay to detect cross-reactive antibodies in serum samples from several bat species in Brazil. Our results warrant further investigation of henipa-like virus reservoirs in the Western hemisphere.

  13. Detection Rate and Clinical Impact of Respiratory Viruses in Children with Kawasaki Disease

    Directory of Open Access Journals (Sweden)

    Ja Hye Kim

    2012-12-01

    Full Text Available &lt;B&gt;Purpose:&lt;/B&gt; The purpose of this prospective case-control study was to survey the detection rate of respiratory viruses in children with Kawasaki disease (KD by using multiplex reverse transcriptasepolymerase chain reaction (RT-PCR, and to investigate the clinical implications of the prevalence of respiratory viruses during the acute phase of KD. &lt;B&gt;Methods:&lt;/B&gt; RT-PCR assays were carried out to screen for the presence of respiratory syncytial virus A and B, adenovirus, rhinovirus, parainfluenza viruses 1 to 4, influenza virus A and B, metapneumovirus, bocavirus, coronavirus OC43/229E and NL63, and enterovirus in nasopharyngeal secretions of 55 KD patients and 78 control subjects. &lt;B&gt;Results:&lt;/B&gt; Virus detection rates in KD patients and control subjects were 32.7% and 30.8%, respectively (P=0.811. However, there was no significant association between the presence of any of the 15 viruses and the incidence of KD. Comparisons between the 18 patients with positive RT-PCR results and the other 37 KD patients revealed no significant differences in terms of clinical findings (including the prevalence of incomplete presentation of the disease and coronary artery diameter. &lt;B&gt;Conclusion:&lt;/B&gt; A positive RT-PCR for currently epidemic respiratory viruses should not be used as an evidence against the diagnosis of KD. These viruses were not associated with the incomplete presentation of KD and coronary artery dilatation.

  14. One-step multiplex real time RT-PCR for the detection of bovine respiratory syncytial virus, bovine herpesvirus 1 and bovine parainfluenza virus 3.

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    Thonur, Leenadevi; Maley, Madeleine; Gilray, Janice; Crook, Tara; Laming, Ellie; Turnbull, Dylan; Nath, Mintu; Willoughby, Kim

    2012-03-28

    Detection of respiratory viruses in veterinary species has traditionally relied on virus detection by isolation or immunofluorescence and/or detection of circulating antibody using ELISA or serum neutralising antibody tests. Multiplex real time PCR is increasingly used to diagnose respiratory viruses in humans and has proved to be superior to traditional methods. Bovine respiratory disease (BRD) is one of the most common causes of morbidity and mortality in housed cattle and virus infections can play a major role. We describe here a one step multiplex reverse transcriptase quantitative polymerase chain reaction (mRT-qPCR) to detect the viruses commonly implicated in BRD. A mRT-qPCR assay was developed and optimised for the simultaneous detection of bovine respiratory syncytial virus (BRSV), bovine herpes virus type 1 (BoHV-1) and bovine parainfluenza virus type 3 (BPI3 i & ii) nucleic acids in clinical samples from cattle. The assay targets the highly conserved glycoprotein B gene of BoHV-1, nucleocapsid gene of BRSV and nucleoprotein gene of BPI3. This mRT-qPCR assay was assessed for sensitivity, specificity and repeatability using in vitro transcribed RNA and recent field isolates. For clinical validation, 541 samples from clinically affected animals were tested and mRT-qPCR result compared to those obtained by conventional testing using virus isolation (VI) and/or indirect fluorescent antibody test (IFAT). The mRT-qPCR assay was rapid, highly repeatable, specific and had a sensitivity of 97% in detecting 102 copies of BRSV, BoHV-1 and BPI3 i & ii. This is the first mRT-qPCR developed to detect the three primary viral agents of BRD and the first multiplex designed using locked nucleic acid (LNA), minor groove binding (MGB) and TaqMan probes in one reaction mix. This test was more sensitive than both VI and IFAT and can replace the aforesaid methods for virus detection during outbreaks of BRD.

  15. Outbreaks of vesicular disease caused by Vaccinia virus in dairy cattle from Goiás State, Brazil (2010-2012

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    Fabiano J.F. de Sant'Ana

    2013-07-01

    Full Text Available Cases of vesicular and exanthematic disease by Vaccinia virus (VACV have been reported in dairy herds of several Brazilian regions, occasionally also affecting humans. The present article describes eight outbreaks of vesicular disease caused by VACV in dairy herds of six counties of Goiás state, Midwestern Brazil (2010-2012, involving a total of 122 cows, 12 calves and 11 people. Dairy cows (3 to 9 years old were affected in all cases and calves (2 to 9 months old were affected in five outbreaks, presenting oral lesions. The morbidity ranged between 8 and 100% in cows, and 1.5 to 31% in calves. In the cows, the clinical signs started with vesicles (2-7mm, painful and coalescent papules (3-8 mm, which resulted in ulcers (5-25mm and scabs in teats, and, occasionally, in the muzzle. The clinical course lasted from 16 to 26 days. The histopathology of bovine skin samples revealed superficial perivascular inflammatory infiltrate of lymphocytes, plasma cells, neutrophils, macrophages and multifocal areas of acanthosis, spongiosis, hipergranulosis and parakeratotic or orthokeratotic hyperkeratosis with adjacent focally extensive ulcers. Eosinophilic inclusion bodies were noted in the cytoplasm of the keratinocytes. PCR to vgf gene of Orthopoxvirus was positive in samples collected from all outbreaks, and in some cases, genomic VACV sequences were identified by nucleotide sequencing of the PCR amplicons. Infectious virus was isolated in cell culture from scabs from one outbreak. Antibodies to Orthopoxvirus were detected in at least 3 or 4 animals in most outbreaks, by ELISA (outbreaks 1, 2, 3, 4, 5 and 7 or virus-neutralization (outbreak 6. Neutralizing titers ranging from 8 to 64 in outbreak 6. In all outbreaks, VACV infection was suspected based on the clinical and pathological findings and it was confirmed by laboratory tests. Upon the etiological confirmation, other agents associated with vesicular disease were discarded. In all outbreaks, at least

  16. Detection of diarrheagenic Escherichia coli from children with and without diarrhea in Salvador, Bahia, Brazil

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    Vanessa Bueris

    2007-11-01

    Full Text Available We identified different diarrheagenic (DEC Escherichia coli pathotypes isolated from 1,207 children with and without acute endemic diarrhea in Salvador, Bahia, Brazil collected as part of a case-control study. Since the identification of DEC cannot be based on only biochemical and culture criteria, we used a multiplex polymerase chain reaction developed by combining five specific primer pairs for Enteropathogenic Escherichia coli (EPEC, Shiga toxin-producing E. coli/ Enterohaemorrhagic E. coli (STEC/EHEC, Enterotoxigenic E. coli (ETEC and Enteroaggregative E. coli (EAEC to detect these pathotypes simultaneously in a single-step reaction. In order to distinguish typical and atypical EPEC strains, these were tested for the presence of EAF plasmid. The prevalence of diarrheagenic E. coli in this sample of a global case-control study was 25.4% (259 patients and 18.7% (35 patients in the diarrhea group (1,020 patients and the control group (187 patients, respectively. The most frequently isolated pathotype was EAEC (10.7%, followed by atypical EPEC (9.4%, ETEC (3.7%, and STEC (0.6%. Typical EPEC was detected only in one sample. The prevalence of the pathotypes studied in children with diarrhea was not significantly different from that in children without diarrhea.

  17. Bovine Papillomavirus in Brazil: Detection of Coinfection of Unusual Types by a PCR-RFLP Method

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    R. F. Carvalho

    2013-01-01

    Full Text Available Bovine papillomavirus (BPV is recognized as a causal agent of benign and malignant tumors in cattle. Thirteen types of BPV are currently characterized and classified into three distinct genera, associated with different pathological outcomes. The described BPV types as well as other putative ones have been demonstrated by molecular biology methods, mainly by the employment of degenerated PCR primers. Specifically, divergences in the nucleotide sequence of the L1 gene are useful for the identification and classification of new papillomavirus types. On the present work, a method based on the PCR-RFLP technique and DNA sequencing was evaluated as a screening tool, allowing for the detection of two relatively rare types of BPV in lesions samples from a six-year-old Holstein dairy cow, chronically affected with cutaneous papillomatosis. These findings point to the dissemination of BPVs with unclear pathogenic potential, since two relatively rare, new described BPV types, which were first characterized in Japan, were also detected in Brazil.

  18. High sensitivity detection and sorting of infectious human immunodeficiency virus (HIV-1) particles by flow virometry.

    Science.gov (United States)

    Bonar, Michał M; Tilton, John C

    2017-05-01

    Detection of viruses by flow cytometry is complicated by their small size. Here, we characterized the ability of a standard (FACSAria II) and a sub-micron flow cytometer (A50 Micro) to resolve HIV-1 viruses. The A50 was superior at resolving small particles but did not reliably distinguish HIV-1, extracellular vesicles, and laser noise by light scatter properties alone. However, single fluorescent HIV-1 particles could readily be detected by both cytometers. Fluorescent particles were sorted and retained infectivity, permitting further exploration of the functional consequences of HIV-1 heterogeneity. Finally, flow cytometry had a limit of detection of 80 viruses/ml, nearly equal to PCR assays. These studies demonstrate the power of flow cytometry to detect and sort viral particles and provide a critical toolkit to validate methods to label wild-type HIV-1; quantitatively assess integrity and aggregation of viruses and virus-based therapeutics; and efficiently screen drugs inhibiting viral assembly and release. Copyright © 2017 Elsevier Inc. All rights reserved.

  19. Development of methodologies for virus detection in soybean and wheat seeds.

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    Botelho, Stephanie R A; Martins, Thais P; Duarte, Macária F; Barbosa, Andreza V; Lau, Douglas; Fernandes, Fernanda R; Sanches, Marcio M

    2016-01-01

    Seeds that contain large amounts of oil, starch, fibers and phenols are the most difficult tissues for RNA extraction. Currently, there are some reports of virus detection in seeds using commercial kits for RNA extraction. However, individual seeds were used, which may not be always suitable for analyses that deal with large amounts of seeds. Sangha [1] described a simple, quick and efficient protocol for RNA extraction and downstream applications in a group of seeds of jatropha (Jatropha curcas), mustard (Brassica sp.) and rice (Oryza sativa). We tested this protocol for soybean (Glycine max), maize (Zea mays), wheat (Triticum aestivum) and triticale (×Triticosecale) seeds and further reverse transcription PCR (RT-PCR)/quantitative real-time PCR (qPCR) in order to have a faster and more practical method for virus detection from seeds than the traditional scheme of seed planting and subsequent Elisa/RT-PCR from leaves. The essential points in the method are:•Some modifications in the protocol [1] were done in order to increase performance: Wheat and triticale seeds are incubated with water prior to maceration. An amount of 1.2 g of dry soybean seeds is used to maceration.•RT-PCR is used for detection of Wheat streak mosaic virus from wheat seeds and RT-qPCR for detection of Soybean mosaic virus from soybean seeds.•The method may be tested for other viruses, however, pre-validation will be needed.

  20. Exhaled breath condensate sampling is not a new method for detection of respiratory viruses

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    Maes Piet

    2011-03-01

    Full Text Available Abstract Background Exhaled breath condensate (EBC sampling has been considered an inventive and novel method for the isolation of respiratory viruses. Methods In our study, 102 volunteers experiencing upper airway infection were recruited over the winter and early spring of 2008/2009 and the first half of the winter of 2009/2010. Ninety-nine EBCs were successfully obtained and screened for 14 commonly circulating respiratory viruses. To investigate the efficiency of virus isolation from EBC, a nasal swab was taken in parallel from a subset of volunteers. The combined use of the ECoVent device with the RTube™ allowed the registration of the exhaled volume and breathing frequency during collection. In this way, the number of exhaled viral particles per liter air or per minute can theoretically be estimated. Results Viral screening resulted in the detection of 4 different viruses in EBC and/or nasal swabs: Rhinovirus, Human Respiratory Syncytial Virus B, Influenza A and Influenza B. Rhinovirus was detected in 6 EBCs and 1 EBC was Influenza B positive. We report a viral detection rate of 7% for the EBCs, which is much lower than the detection rate of 46.8% observed using nasal swabs. Conclusion Although very promising, EBC collection using the RTube™ is not reliable for diagnosis of respiratory infections.

  1. High rates of double-stranded RNA viruses and Mycoplasma hominis in Trichomonas vaginalis clinical isolates in South Brazil.

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    da Luz Becker, Débora; dos Santos, Odelta; Frasson, Amanda Piccoli; de Vargas Rigo, Graziela; Macedo, Alexandre José; Tasca, Tiana

    2015-08-01

    Trichomonas vaginalis is the etiological agent of trichomoniasis, the most common non-viral sexually transmitted disease (STD) in world, with 276.4 million new cases each year. T. vaginalis can be naturally infected with Mycoplasma hominis and Trichomonasvirus species. This study aimed to evaluate the prevalence of T. vaginalis infected with four distinct T. vaginalis viruses (TVVs) and M. hominis among isolates from patients in Porto Alegre city, South Brazil. An additional goal of this study was to investigate whether there is association between metronidazole resistance and the presence of M. hominis during TVV infection. The RNA expression level of the pyruvate ferredoxin oxidoreductase (PFOR) gene was also evaluated among metronidazole-resistant and metronidazole-sensitive T. vaginalis isolates. A total of 530 urine samples were evaluated, and 5.7% samples were positive for T. vaginalis infection. Among them, 4.51% were isolated from female patients and 1.12% were from male patients. Remarkably, the prevalence rates of M. hominis and TVV-positive T. vaginalis isolates were 56.7% and 90%, respectively. Most of the T. vaginalis isolates were metronidazole-sensitive (86.7%), and only four isolates (13.3%) were resistant. There is no statistically significant association between infection by M. hominis and infection by TVVs. Our results refute the hypothesis that the presence of the M. hominis and TVVs is enough to confer metronidazole resistance to T. vaginalis isolates. Additionally, the role of PFOR RNA expression levels in metronidazole resistance as the main mechanism of resistance to metronidazole could not be established. This study is the first report of the T. vaginalis infection by M. hominis and TVVs in a large collection of isolates from South Brazil. Copyright © 2015 Elsevier B.V. All rights reserved.

  2. Hepatitis virus and hepatocellular carcinoma in Brazil: a report from the State of Espírito Santo

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    Patrícia Lofêgo Gonçalves

    2014-10-01

    Full Text Available Introduction Few studies have examined hepatocellular carcinoma (HCC in Brazil, and the incidence and risk factors for this type of malignancy vary greatly geographically. In this paper, we report several risk factors associated with HCC diagnosed at the University Hospital in Vitória, ES, Brazil. Methods We reviewed 274 cases of HCC (January 1993 to December 2011 in which hepatitis B (HBV and C (HCV virus infection and chronic alcoholism were investigated. A diagnosis of hepatocellular carcinoma was confirmed by histology or by the presence of a characteristic pattern on imaging. Results HCC with associated liver cirrhosis was noted in 85.4% of cases. The mean ages of men and women were 56.6 years and 57.5 years, respectively. The male-to-female ratio was 5.8:1. Associated risk factors included the following: HBV, 37.6% (alone, 23.4%; associated with chronic alcoholism, 14.2%; HCV, 22.6% (alone, 13.5%; associated with chronic alcoholism, 9.1%, chronic alcoholism, 17.1%, non-alcoholic steatohepatitis, 2.6% and cryptogenic, 19.3%. The male-to-female ratio was higher in cases associated with HBV or chronic alcoholism compared with HCV-associated or cryptogenic cases. In 40 cases without associated cirrhosis, the male-to-female ratio and mean age were lower than those in cirrhosis-associated cases. Conclusions These results demonstrate that the main risk factor associated with HCC in the State of Espírito Santo is HBV. Chronic alcoholism is an important etiological factor, alone or in association with HBV or HCV infection.

  3. HIV behind bars: human immunodeficiency virus cluster analysis and drug resistance in a reference correctional unit from southern Brazil.

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    Isabel M Prellwitz

    Full Text Available People deprived of liberty in prisons are at higher risk of infection by the human immunodeficiency virus (HIV due to their increased exposure through intravenous drug use, unprotected sexual activity, tattooing in prison and blood exposure in fights and rebellions. Yet, the contribution of intramural HIV transmission to the epidemic is scarcely known, especially in low- and middle-income settings. In this study, we surveyed 1,667 inmates incarcerated at Presídio Central de Porto Alegre, located in southern Brazil, for HIV infection and molecular characterization. The HIV seroprevalence was 6.6% (110/1,667. Further analyses were carried out on 40 HIV-seropositive inmates to assess HIV transmission clusters and drug resistance within the facility with the use of molecular and phylogenetic techniques. The molecular epidemiology of HIV-1 subtypes observed was similar to the one reported for the general population in southern Brazil, with the predominance of HIV-1 subtypes C, B, CRF31_BC and unique BC recombinants. In particular, the high rate (24% of URF_BC found here may reflect multiple exposures of the population investigated to HIV infection. We failed to find HIV-infected inmates sharing transmission clusters with each other. Importantly, the analysis of HIV-1 pol genomic fragments evidenced high rates of HIV primary and secondary (acquired drug resistance and an alarming proportion of virologic failure among patients under treatment, unveiling suboptimal access to antiretroviral therapy (ARV, low ARV adherence and dissemination of drug resistant HIV strains in primary infections. Our results call for immediate actions of public authority to implement preventive measures, serological screening and, for HIV-seropositive subjects, clinical and treatment follow-up in order to control HIV infection and limit the spread of drug resistance strains in Brazilian prisons.

  4. Simultaneous detection of Hepatitis B virus and Hepatitis C virus in human plasma using Taq-man chemistry

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    Khaja M N

    2011-07-01

    Full Text Available Designing a rapid, reliable and sensitive assay, for detection of hepatitis B virus and Hepatitis C virus variants by real-time PCR, is challenging at best. A recent approach for quantifying the viral load using the sensitive fluorescence principle, was used in this study. A total of 350 samples were collected from outpatient unit, Center for Liver Research and Diagnostics (CLRD. Complete Human HBV DNA and HCV sequences were obtained from the National Centre for Biotechnology Information (NCBI; primers and probes were designed and synthesized from core, surface and x region of Hepatitis B and UTR region of HCV. Real-time based detection was done, using standard kit and in-house generated standards and RT-PCR protocols. A standard curve was generated by using the Smart Cycler II software and serial dilution 102 to 108 of cloned viral regions, the calibration curve was linear in a range from 102 to108 cp/ml for both HBV and HCV, with R2 value of 0.999 and 0.995. Out of 100 predetermined HCV negative samples, 02 samples were found positive with in-house developed RT-PCR assay, the positivity of this sample was confirmed by sequencing the amplified product. Low cost of this assay procedure and précised sample volume will permit the assay to be implemented for routine screening of Hepatitis B and C virus mono-infection and co-infection using Real Time PCR , Nucleic acid Chip technology and Fluorescent End Point detection systems. This assay is reproducible showing limited inter and intra assay variability. Our results correlated well with the standard kit for HBV and HCV virus monitor.

  5. First molecular detection of Aichi virus in sewage and shellfish samples in the Monastir region of Tunisia.

    Science.gov (United States)

    Sdiri-Loulizi, K; Hassine, M; Aouni, Z; Gharbi-Khelifi, H; Sakly, N; Chouchane, S; Guédiche, M N; Pothier, P; Aouni, M; Ambert-Balay, K

    2010-09-01

    The aims of our investigations were (1) to look for Aichi virus in environmental samples and (2) to compare the Aichi virus strains in both clinical and environmental samples in order to evaluate the role of environmental contamination as a possible vehicle for viral transmission. Aichi virus was detected in 15 (6%) sewage samples and in 4 (6.6%) shellfish samples. Aichi virus was identified for the first time in water samples. Phylogenetic analysis revealed several clusters that occurred sequentially in time, suggesting some parallelism in the evolution of environmental and human strains. Aichi virus present in sewage reflects the viruses circulating in the community.

  6. Health and Development at Age 19-24 Months of 19 Children Who Were Born with Microcephaly and Laboratory Evidence of Congenital Zika Virus Infection During the 2015 Zika Virus Outbreak - Brazil, 2017.

    Science.gov (United States)

    Satterfield-Nash, Ashley; Kotzky, Kim; Allen, Jacob; Bertolli, Jeanne; Moore, Cynthia A; Pereira, Isabela Ornelas; Pessoa, André; Melo, Flavio; Santelli, Ana Carolina Faria E Silva; Boyle, Coleen A; Peacock, Georgina

    2017-12-15

    In November 2015, the Brazilian Ministry of Health (MOH) declared the Zika virus outbreak a public health emergency after an increase in microcephaly cases was reported in the northeast region of the country (1). During 2015-2016, 15 states in Brazil with laboratory-confirmed Zika virus transmission reported an increase in birth prevalence of microcephaly (2.8 cases per 10,000 live births), significantly exceeding prevalence in four states without confirmed transmission (0.6 per 10,000) (2). Although children with microcephaly and laboratory evidence of Zika virus infection have been described in early infancy (3), their subsequent health and development have not been well characterized, constraining planning for the care and support of these children and their families. The Brazilian MOH, the State Health Secretariat of Paraíba, and CDC collaborated on a follow-up investigation of the health and development of children in northeastern Brazil who were reported to national surveillance with microcephaly at birth. Nineteen children with microcephaly at birth and laboratory evidence of Zika virus infection were assessed through clinical evaluations, caregiver interviews, and review of medical records. At follow-up (ages 19-24 months), most of these children had severe motor impairment, seizure disorders, hearing and vision abnormalities, and sleep difficulties. Children with microcephaly and laboratory evidence of Zika virus infection have severe functional limitations and will require specialized care from clinicians and caregivers as they age.

  7. Field-Usable Lateral Flow Immunoassay for the Rapid Detection of White Spot Syndrome Virus (WSSV).

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    Kulabhusan, Prabir Kumar; Rajwade, Jyutika M; Sugumar, Vimal; Taju, Gani; Sahul Hameed, A S; Paknikar, Kishore M

    2017-01-01

    White spot disease (WSD), a major threat to sustainable aquaculture worldwide, is caused by White spot syndrome virus (WSSV). The diagnosis of WSD relies heavily on molecular detection of the virus by one-step PCR. These procedures are neither field-usable nor rapid enough considering the speed at which the virus spreads. Thus, development of a rapid, reliable and field-usable diagnostic method for the detection of WSSV infection is imperative to prevent huge economic losses. Here, we report on the development of a lateral flow immunoassay (LFIA) employing gold nanoparticles conjugated to a polyclonal antibody against VP28 (envelope protein of WSSV). The LFIA detected WSSV in ~20 min and showed no cross-reactivity with other shrimp viruses, viz. Monodon Baculovirus (MBV), Hepatopancreatic parvovirus (HPV) and Infectious Hypodermal and Hematopoietic Necrosis virus (IHHNV). The limit of detection (LOD) of the assay, as determined by real-time PCR, was 103 copies of WSSV. In a time course infectivity experiment, ~104 WSSV particles were injected in Litopenaeus vannamei. The LFIA could rapidly (~ 20 min) detect the virus in different tissues after 3 h (hemolymph), 6 h (gill tissue) and 12 h (head soft tissue, eye stalk, and pleopod) of infection. Based on these findings, a validation study was performed using 75 field samples collected from different geographical locations in India. The LFIA results obtained were compared with the conventional "gold standard test", viz. one-step PCR. The analysis of results in 2x2 matrix indicated very high sensitivity (100%) and specificity (96.77%) of LFIA. Similarly, Cohen's kappa coefficient of 0.983 suggested "very good agreement" between the developed LFIA and the conventional one-step PCR. The LFIA developed for the rapid detection of WSSV has an excellent potential for use in the field and could prove to be a boon to the aquaculture industry.

  8. Detection of influenza A virus in live bird markets in Kenya, 2009-2011.

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    Munyua, Peninah M; Githinji, Jane W; Waiboci, Lilian W; Njagi, Leonard M; Arunga, Geoffrey; Mwasi, Lydia; Murithi Mbabu, R; Macharia, Joseph M; Breiman, Robert F; Kariuki Njenga, M; Katz, Mark A

    2013-03-01

    Surveillance for influenza viruses within live bird markets (LBMs) has been recognized as an effective tool for detecting circulating avian influenza viruses (AIVs). In Sub-Saharan Africa, limited data exist on AIVs in animal hosts, and in Kenya the presence of influenza virus in animal hosts has not been described. This surveillance project aimed to detect influenza A virus in poultry traded in five LBMs in Kenya. We visited each market monthly and collected oropharyngeal and cloacal specimens from poultry and environmental specimens for virological testing for influenza A by real time RT-PCR. On each visit, we collected information on the number and types of birds in each market, health status of the birds, and market practices. During March 24, 2009-February 28, 2011, we collected 5221 cloacal and oropharyngeal swabs. Of the 5199 (99·6%) specimens tested, influenza A virus was detected in 42 (0·8%), including 35/4166 (0·8%) specimens from chickens, 3/381 (0·8%) from turkeys, and 4/335 (1·2%) from geese. None of the 317 duck specimens were positive. Influenza was more commonly detected in oropharyngeal [33 (1·3%)] than in cloacal [9 (0·4%)] specimens. None of the 485 environmental specimens were positive. Virus was detected in all five markets during most (14/22) of the months. Ducks and geese were kept longer at the market (median 30 days) than chickens (median 2days). Influenza A was detected in a small percentage of poultry traded in LBMs in Kenya. Efforts should be made to promote practices that could limit the maintenance and transmission of AIVs in LBMs. © 2012 Blackwell Publishing Ltd.

  9. Detection of beet soil-borne virus and beet virus Q in sugarbeet in Greece

    NARCIS (Netherlands)

    Pavli, R.; Prins, M.; Skaracis, G.N.

    2010-01-01

    Sugar beet plants with typical rhizomania symptoms were collected from the five major cultivation zones of Greece. The presence of Beet necrotic yellow vein virus (BNYVV), the primary causal agent of the disease, was ascertained by DAS-ELISA in 38 out of 40 fields surveyed and the positive samples

  10. Respiratory syncytial virus evaluation among asymptomatic and symptomatic subjects in a university hospital in Sao Paulo, Brazil in the period of 2009 to 2013.

    Science.gov (United States)

    Moreira, Luciana Peniche; Watanabe, Aripuana Sakurada Aranha; Camargo, Clarice Neves; Melchior, Thais Boim; Granato, Celso; Bellei, Nancy

    2017-10-27

    RSV is recognized as an important cause of respiratory tract infections. Immunocompromised patients, health care workers (HCW) and children contacts are at increased risk of acquiring the infection. However, the impact of asymptomatic infection in transmission has not been well studied. This study evaluated the frequency and viral load (VL) of RSV in nasal swab samples of individuals with different risk factors for acquiring infection in a university hospital in Sao Paulo, Brazil. We included 196 symptomatic children and their 192 asymptomatic caregivers, 70 symptomatic and 95 asymptomatic HCW, 43 samples from symptomatic HIV positive outpatients and 100 samples of asymptomatic HIV patients in the period of 2009 to 2013. RSV infection was detected in 10.1% (70/696) of samples, 4.4% (17/387) of asymptomatic patients and 17.1% (53/309) from symptomatic patients. (p< 0.0001). The VL of symptomatic patients (4.7 log copies/mL) was significantly higher compared to asymptomatic patients (2.3 log copies/mL). RSV detection among asymptomatic caregivers (6.8%; 13/192) was significantly higher compared to other asymptomatic adults, HIV and HCWS (2.0%; 4/195; p = 0.0252). A close contact with an infected child at home was an important risk to RSV acquisition [OR 22.6 (95% CI 4.8 to 106.7)]. Children who possibly transmitted the virus to their asymptomatic contacts had significantly higher viral load than children who probably did not transmit (p < 0.0001). This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  11. Infections by Leptospira interrogans, Seoul virus, and Bartonella spp. among Norway rats (Rattus norvegicus) from the urban slum environment in Brazil.

    Science.gov (United States)

    Costa, Federico; Porter, Fleur Helena; Rodrigues, Gorete; Farias, Helena; de Faria, Marcus Tucunduva; Wunder, Elsio A; Osikowicz, Lynn M; Kosoy, Michael Y; Reis, Mitermayer Galvão; Ko, Albert I; Childs, James E

    2014-01-01

    Norway rats (Rattus norvegicus) are reservoir hosts for zoonotic pathogens that cause significant morbidity and mortality in humans. Studies evaluating the prevalence of zoonotic pathogens in tropical Norway rat populations are rare, and data on co-infection with multiple pathogens are nonexistent. Herein, we describe the prevalence of leptospiral carriage, Seoul virus (SEOV), and Bartonella spp. infection independently, in addition to the rates of co-infection among urban, slum-dwelling Norway rats in Salvador, Brazil, trapped during the rainy season from June to August of 2010. These data were complemented with previously unpublished Leptospira and SEOV prevalence information collected in 1998. Immunofluorescence staining of kidney impressions was used to identify Leptospira interrogans in 2010, whereas isolation was used in 1998, and western blotting was used to detect SEOV antibodies in 2010, whereas enzyme-linked immunosorbent assay (ELISA) was used in 1998: in 2010, Bartonella spp. were isolated from a subsample of rats. The most common pathogen in both years was Leptospira spp. (83%, n=142 in 1998, 63%, n=84 in 2010). SEOV was detected in 18% of individuals in both 1998 and 2010 (n=78 in 1998; n=73 in 2010), and two species of Bartonella were isolated from 5 of 26 rats (19%) tested in 2010. The prevalence of all agents increased significantly with rat mass/age. Acquisition of Leptospira spp. occurred at a younger mass/age than SEOV and Bartonella spp. infection, suggesting differences in the transmission dynamics of these pathogens. These data indicate that Norway rats in Salvador serve as reservoir hosts for all three of these zoonotic pathogens and that the high prevalence of leptospiral carriage in Salvador rats poses a high degree of risk to human health.

  12. A quantitative real-time reverse transcription PCR (qRT-PCR) assay to detect genome segment 9 of all 26 bluetongue virus serotypes.

    Science.gov (United States)

    Maan, Narender S; Maan, Sushila; Belaganahalli, Manjunatha; Pullinger, Gillian; Montes, Antonio J Arenas; Gasparini, Marcela R; Guimera, Marc; Nomikou, Kyriaki; Mertens, Peter P C

    2015-03-01

    Bluetongue (BT) is an arboviral disease, which can often be fatal in naïve sheep and white tailed deer, but is usually less severe, or unapparent in other ruminants. Twenty-six bluetongue virus (BTV) serotypes have been recognised so far, two of which (BTV-25 and BTV-26) were recently identified by phylogenetic comparisons of genome-segment/outer-capsid protein VP2 (subsequently confirmed by serological 'virus-neutralisation' assays). Rapid, sensitive, reliable and quantitative diagnostic-assays for detection and identification of BTV represent important components of effective surveillance and control strategies. The BTV genome comprises 10 linear segments of dsRNA. We describe a 'TaqMan' fluorescence-probe based quantitative real-time RT-PCR assay, targeting the highly conserved genome-segment-9 (encoding the viral-helicase 'VP6' and NS4). The assay detected Seg-9 from isolates of all 26 BTV types, as well as from clinical samples derived from BTV-6w and BTV-8w outbreaks (in Europe), BTV-25 from Switzerland, BTV-26 from Kuwait, BTV-1w, BTV-4w and BTV-8w from Spain, BTV-4w, BTV-8, BTV-10 and BTV-16 from Brazil. Assay efficiency was evaluated with RNA derived from the reference strain of BTV-1w [RSArrrr/01] and was 99.6%, detecting down to 4 copies per reaction. Samples from uninfected insect or mammalian cell-cultures, hosts-species (uninfected sheep blood) or vector-insects, all gave negative results. The assay failed to detect RNA from heterologous but related Orbivirus species (including the nine African horse sickness virus [AHSV] and seven epizootic haemorrhagic disease virus [EHDV] serotypes). Copyright © 2014. Published by Elsevier B.V.

  13. Anticorpos contra o vírus da língua azul em bovinos do sertão da Paraíba Antibodies to bluetongue virus in bovines of Paraíba State, Brazil

    Directory of Open Access Journals (Sweden)

    C.B. Melo

    2000-02-01

    Full Text Available In June of 1997 the prevalence of antibodies to bluetongue virus was between 3.94 and 4.82% in 137 bovine serum samples from 12 herds in Paraiba State, Brazil. This is the first report of antibodies to bluetongue virus in Paraiba State herds.

  14. Detection of Quang Binh virus from mosquitoes in China.

    Science.gov (United States)

    Zuo, Shuqing; Zhao, Qiumin; Guo, Xiaofang; Zhou, Hongning; Cao, Wuchun; Zhang, Jiusong

    2014-02-13

    Flaviviruses present a wide range of genetic diversity and exhibit diverse host relationships. Mosquito-borne flaviviruses have recently been isolated and characterized worldwide. Yunnan Province of China is one of the richest areas of species diversity and is the center of multi-species evolution in mainland Asia, which supports the circulation of numerous arthropod-borne viruses (arboviruses). In a screening program of arboviruses, mosquitoes were collected during the mosquito activity season in the Yunnan Province from 2007 to 2010. Eleven flavivirus strains, named Yunnan Culex flaviviruses (YNCxFVs), were obtained from Culex tritaeniorhynchus and Anopheles sinensis specimens. Sequence analyses based on partial nonstructural protein (NS) 5 gene indicated that the YNCxFVs shared 92.8-99.6% nucleotide identity with each other and were similar to the Culex-related flaviviruses. The complete genome of one representative isolate, LSFlaviV-A20-09, was sequenced. The genome was 10,865 nucleotides long and contained a single, long open reading frame (ORF) of 10,080 nucleotides that encoded a 3360-aa polyprotein. This genome was most closely related to the Quang Binh virus (QBV) VN180 strain, an insect-specific flavivirus isolated from Culex mosquitoes in Vietnam, but only had 83.0% nucleotide and 93.8% amino acid identities for the ORF sequence. The genome has approximately 66.3%-68.5% nucleotide sequence and 69.3-73.3% amino acid sequence identities to other Culex flaviviruses, and only has 47.9-57.9% nucleotide sequence and 38.7-55.1% amino acid sequence identities to Coquillettidia-related, Mansonia-related and Aedes-related flaviviruses. Phylogenetic analyses revealed that the LSFlaviV-A20-09 fell into the Culex-related flavivirus clade. Our discoveries provide more information regarding the heterogeneity of viruses that infect mosquitoes. Copyright © 2013 Elsevier B.V. All rights reserved.

  15. Genotyping of hepatitis B virus in a cohort of patients evaluated in a hospital of Porto Alegre, south of Brazil

    Directory of Open Access Journals (Sweden)

    Carlos Eduardo Becker

    2010-03-01

    Full Text Available CONTEXT: In recent years the hepatitis B virus (HBV genotyping has been considered a relevant factor in the natural history of the disease. OBJECTIVE: To determine hepatitis B virus genotypes and its epidemiological and clinical implications, in a cohort of patients in a hospital in Porto Alegre, South of Brazil. Methods - Sixty seven patients with HBV chronic infection markers who were being treated at ''Complexo Hospitalar Santa Casa'', in Porto Alegre, RS, Brazil, were evaluated. Demographic and epidemiological data were collected from these group of patients by following a standard protocol and ALT and HBeAg were determined. The genotypes and subtypes were determined by in-house PCR and, finally, the samples were sequenced. The level of significance used was 5%. RESULTS: The qualitative analysis for HBV-DNA by PCR was positive in 79.1% of the samples (53/67. The genotype was determined in all positive VHB-DNA samples and the genotypes A (34%, D (60.4% and F (5.4% as well as the subtypes adw, ayw and adw4 were found. No significant correlation was found between the hepatitis B virus genotypes and demographic variables considered as risk factors for hepatitis B virus infection. There was also no correlation between the genotypes and the serological and laboratory variables related to liver disease. CONCLUSION: We concluded that the most prevalent genotype found was D. However, further studies are needed to allow us to evaluate the implications of genetic variability in the clinical evolution of HBV carriers.CONTEXTO: Nos últimos anos a genotipagem do vírus da hepatite B (VHB tem sido considerado fator relevante para a história natural da doença. OBJETIVOS: Determinar os genótipos do VHB e suas implicações clínicas e epidemiológicas, em uma coorte de pacientes em um hospital de Porto Alegre, RS, sul do Brasil. MÉTODOS: Foram avaliados 67 pacientes com marcadores de infecção crônica pelo VHB que estavam sendo tratados no Complexo

  16. Flow cytometric detection of viruses in the Zuari estuary, Goa

    Digital Repository Service at National Institute of Oceanography (India)

    Mitbavkar, S.; Rajaneesh, K.M.; SathishKumar, P.

    diluted in 0.2 μm filtered, auto- claved TE-buffer (pH 8.0) and stained with SYBR Green I (final conc. 0.5 × 10 –4 of the commercial stock) for 10 min at 80°C in the dark, followed by a cool- ing period of 5 min at room temperature before analysis 10... . Based on their relative green fluores- cence and side scatter signatures, two virus subpopulations were distinguished, defined as V1 and V2, corresponding to populations observed previously in sea- water samples 11 (Figure 1). The viral...

  17. Detection of antibodies against capripoxviruses using an inactivated sheeppox virus ELISA.

    Science.gov (United States)

    Babiuk, S; Wallace, D B; Smith, S J; Bowden, T R; Dalman, B; Parkyn, G; Copps, J; Boyle, D B

    2009-05-01

    An indirect ELISA was developed to detect antibodies specific for capripoxviruses in goat, sheep and cattle sera. Heat-inactivated Nigerian sheeppox virus was used as the ELISA antigen. Sera obtained from sheep and goats that were experimentally infected with different capripoxvirus isolates were used to develop and evaluate the sensitivity of the ELISA. Virus neutralization indexes were determined for the experimental sera in OA3.Ts cells. The specificity of the ELISA was determined using 231 sera from capripoxvirus naïve sheep and goats from Canada. In addition, the ELISA was tested for cross-reactivity to anti-orf virus antibodies using orf-reactive sera and no cross-reactivity was observed. Using experimentally generated sera obtained from animals infected with virulent sheeppox or goatpox virus isolates, the diagnostic sensitivity of the ELISA was 96% with a diagnostic specificity of 95%, where the diagnostic sensitivity of the virus neutralization assay was 96% with a diagnostic specificity of 100%. Further evaluation of this ELISA, using 276 cattle serum samples that were positive by virus neutralization assays, revealed a diagnostic sensitivity of 88% with a specificity of 97%. These results indicated that the inactivated capripoxvirus ELISA can detect capripoxvirus-specific antibodies in sheep, goats and cattle that have been infected with virulent capripoxvirus isolates. Non-virulent capripoxvirus isolates, in contrast, did not elicit positive (>or=1.5 Log10 neutralization index) antibody responses.

  18. Timing of First Respiratory Virus Detections in Infants: A Community-Based Birth Cohort Study.

    Science.gov (United States)

    Sarna, Mohinder; Ware, Robert S; Lambert, Stephen B; Sloots, Theo P; Nissen, Michael D; Grimwood, Keith

    2018-01-17

    Determining timing of first virus detection episodes (fVDEs) for different respiratory viruses in infants identifies risk periods and informs preventive interventions, including vaccination. We describe the ages and nature of fVDEs in an infant birth cohort and explore factors associated with increased odds of symptomatic fVDEs. The Observational Research in Childhood Infectious Diseases (ORChID) study is a community-based birth cohort describing acute respiratory infections in infants until their second birthday. Parents recorded daily symptoms and collected nose swabs weekly, which were batch-tested using polymerase chain reaction assays for 17 respiratory viruses. One hundred fifty-eight infants participated in ORChID. The median age for fVDEs was 2.9 months for human rhinovirus (HRV) but was ≥13.9 months for other respiratory viruses. Overall, 52% of HRV fVDEs were symptomatic, compared with 57%-83% of other fVDEs. Respiratory syncytial virus and human metapneumovirus fVDEs were more severe than HRV fVDEs. Older age and the winter season were associated with symptomatic episodes. Infants do not always experience respiratory symptoms with their fVDE. Predominance of early HRV detections highlights the need for timing any intervention early in life. fVDEs from other respiratory viruses most commonly occur when maternal vaccines may no longer provide protection.

  19. Rapid detection of avian influenza A virus by immunochromatographic test using a novel fluorescent dye.

    Science.gov (United States)

    Yeo, Seon-Ju; Cuc, Bui Thi; Kim, Soon-Ai; Kim, Do Thi Hoang; Bao, Duong Tuan; Tien, Trinh Thi Thuy; Anh, Nguyen Thi Viet; Choi, Do-Young; Chong, Chom-Kyu; Kim, Hak Sung; Park, Hyun

    2017-08-15

    Sensitive and rapid diagnostic systems for avian influenza (AI) virus are required to screen large numbers of samples during a disease outbreak and to prevent the spread of infection. In this study, we employed a novel fluorescent dye for the rapid and sensitive recognition of AI virus. The styrylpyridine phosphor derivative was synthesized by adding allyl bromide as a stable linker and covalently immobilizing it on latex beads with antibodies generating the unique Red dye 53-based fluorescent probe. The performance of the innovative rapid fluorescent immnunochromatographic test (FICT) employing Red dye 53 in detecting the AI virus (A/H5N3) was 4-fold and 16-fold higher than that of Europium-based FICT and the rapid diagnostic test (RDT), respectively. In clinical studies, the presence of human nasopharyngeal specimens did not alter the performance of Red dye 53-linked FICT for the detection of H7N1 virus. Furthermore, in influenza A virus-infected human nasopharyngeal specimens, the sensitivity of the Red dye 53-based assay and RDT was 88.89% (8/9) and 55.56% (5/9) relative to rRT-PCR, respectively. The photostability of Red dye 53 was higher than that of fluorescein isothiocyanate (FITC), showing a stronger fluorescent signal persisting up to 8min under UV. The Red dye 53 could therefore be a potential probe for rapid fluorescent diagnostic systems that can recognize AI virus in clinical specimens. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. Detection of Leishmania infantum and a Novel Phlebovirus (Balkan Virus) from Sand Flies in Albania.

    Science.gov (United States)

    Ayhan, Nazli; Velo, Enkelejda; de Lamballerie, Xavier; Kota, Majlinda; Kadriaj, Perparim; Ozbel, Yusuf; Charrel, Remi N; Bino, Silvia

    2016-12-01

    To organize entomological campaigns to trap sand flies in selected regions of Albania and to test them for the presence of existing or new phleboviruses and for leishmania DNA. Sand flies were collected in 14 locations from May to October 2014 using three different types of traps. Pools with a maximum of 30 individuals were prepared according to gender, trapping site, and trapping date; they were tested for the presence of (1) phlebovirus RNA with three different PCR systems (2) and Leishmania DNA using two different real-time PCR assays. A total of 972 sand flies (568 females, 404 males) were aliquoted to 55 pools. Three pools (in two different regions) were positive for Leishmania infantum. Two pools (Kruje region) were positive for phlebovirus RNA and a 575-nucleotide (nt) colinearized sequence of a novel virus most closely related to but clearly distinct from Tehran virus (16% and 3% divergence at nt and amino acid levels). Next generation sequencing analysis indicated that this virus might be transmitted by either Phlebotomus neglectus, Phlebotomus tobbi, or both vectors. Visceral leishmaniasis has been clinically recognized in Albania for at least 80 years; however, this is the first time that L. infantum, detected by molecular means, has been reported in sand flies in Albania. At the outset of this study, only Adria virus (Salehabad species) was recognized in Albania. A novel virus, Balkan virus, was identified and genetic analysis revealed that it belongs to the Sandfly fever Naples virus group containing human pathogens.

  1. Detection and molecular characterization of enteric viruses from poult enteritis syndrome in turkeys.

    Science.gov (United States)

    Jindal, N; Patnayak, D P; Chander, Y; Ziegler, A F; Goyal, S M

    2010-02-01

    This study was conducted to detect and characterize enteric viruses [rotavirus, turkey astrovirus-2 (TAstV-2), reovirus, and turkey coronavirus] from cases of poult enteritis syndrome (PES) in Minnesota turkeys. Of the intestinal contents collected from 43 PES cases, 25 were positive for rotavirus and 13 for small round viruses by electron microscopy (EM). Of the enteric virus-positive cases by EM (n=27), 16 cases had rotavirus or small round viruses alone and the remaining 11 cases had both viruses. None of the cases were positive for reovirus or coronavirus by EM. However, with reverse transcription-PCR (RT-PCR), 40 cases (93%) were positive for rotavirus, 36 (84%) for TAstV-2, and 17 (40%) for reovirus. None of the cases were positive for turkey coronavirus by RT-PCR. The viruses from all cases were detected either alone or in combination of 2 or 3 by RT-PCR. Thus, 8 (19%) cases were positive for a single virus, whereas a combination of viruses was detected in the remaining 35 (81%) cases. The rota-TAstV-2 combination was the most predominant (n=18 cases). Fifteen cases were positive for all 3 viruses. The rotaviruses had sequence homology of 89.8 to 100% with previously published sequences of turkey rotaviruses at the nucleotide level. The TAstV-2 had sequence homology of 84.6 to 98.7% with previously published TAstV-2, whereas reoviruses had sequence homology of 91.6 to 99.3% with previously published sequences of turkey reoviruses. Phylogenetic analysis revealed that rota- and reoviruses clustered in a single group, whereas TAstV-2 clustered in 2 different groups. In conclusion, a larger number of PES cases was positive for rotavirus, TAstV-2, and reovirus by RT-PCR than with EM. The presence of more than one virus and changes at the genetic level in a virus may affect the severity of PES in turkey flocks.

  2. New molecular methods for the detection of hepatitis A and Norwalk viruses in shellfish.

    Science.gov (United States)

    Romalde, J L

    1996-12-01

    Outbreaks of viral enteric diseases after consumption of shellfish are a major health risk. Methodological problems (such as toxicity for cell cultures and low viral concentrations) and the unculturability of some strains (i.e. hepatitis A virus, Norwalk virus) have made it difficult to study those viruses in the environmental samples. Currently, the analysis of the hygienic quality of marketable shellfish is determined by the use of fecal indicator bacteria, but their reliability in determining viral pollution of shellfish is very low. Recent biotechnology developments are providing available rapid, sensitive, and specific tools for detecting food-borne viruses in shellfish and in shellfish-growing waters. In this paper, a review of these new molecular methods is carried out, discussing their advantages and possible applications.

  3. Henipavirus microsphere immuno-assays for detection of antibodies against Hendra virus.

    Science.gov (United States)

    McNabb, Leanne; Barr, J; Crameri, G; Juzva, S; Riddell, S; Colling, A; Boyd, V; Broder, C; Wang, L-F; Lunt, R

    2014-05-01

    Hendra and Nipah viruses (HeV and NiV) are closely related zoonotic pathogens of the Paramyxoviridae family. Both viruses belong to the Henipavirus genus and cause fatal disease in animals and humans, though only HeV is endemic in Australia. In general and due to the acute nature of the disease, agent detection by PCR and virus isolation are the primary tools for diagnostic investigations. Assays for the detection of antibodies against HeV are fit more readily for the purpose of surveillance testing in disease epidemiology and to meet certification requirements in the international movement of horses. The first generation indirect ELISA has been affected by non-specific reactions which must be resolved using virus neutralisation serology conducted at laboratory bio-safety level 4 containment (PC4). Recent developments have enabled improvements in the available serology assays. The production of an expressed recombinant truncated HeV G protein has been utilised in ELISA and in Luminex-based multiplexed microsphere assays. In the latter format, two Luminex assays have been developed for use in henipavirus serology: a binding assay (designed for antibody detection and differentiation) and a blocking assay (designed as a surrogate for virus neutralisation). Equine and canine field sera were used to evaluate the two Luminex assays relative to ELISA and virus neutralisation serology. Results showed that Luminex assays can be effective as rapid, sensitive and specific tests for the detection of HeV antibody in horse and dog sera. The tests do not require PC4 containment and are appropriate for high throughput applications as might be required for disease investigations and other epidemiological surveillance. Also, the results show that the Luminex assays detect effectively HeV vaccine-induced antibodies. Copyright © 2014 Elsevier B.V. All rights reserved.

  4. Comparison of sputum and nasopharyngeal swabs for detection of respiratory viruses.

    Science.gov (United States)

    Jeong, Ji Hun; Kim, Kyung Hee; Jeong, Sung Hwan; Park, Jeong Woong; Lee, Sang Min; Seo, Yiel Hea

    2014-12-01

    Diagnostic tests for respiratory viral infections use traditionally either nasopharyngeal washes or swabs. Sputum is representative of the lower respiratory tract but is used rarely for viral testing. The aim of this study was to compare the detection rates of respiratory viruses from nasopharyngeal swabs and sputum using a multiplex real-time reverse transcription-polymerase chain reaction (RT-PCR). Adults who were admitted or presented to the clinics of Gil Medical Center with acute respiratory symptoms were recruited from 1 November 2012 to 31 March 2013. Paired specimens of nasopharyngeal swabs and sputum were obtained from 154 subjects, and RNA was extracted and tested for 16 different respiratory viruses using the Anyplex II RV16 Detection kit (Seegene, Seoul, Korea). The positive rate was 53% (81/154) for nasopharyngeal swabs and 68% (105/154) for sputum (P < 0.001). One hundred thirty-four viruses were identified for 107 illnesses. Influenza A virus, RSV A, HRV, coronavirus OC43, and adenovirus were detected more frequently in sputum samples than in nasopharyngeal swabs (P < 0.001). Importantly, 12 of 44 (27%) influenza A infections and 11 of 27 (41%) RSV infections were positive in only sputum samples. The detection rates of respiratory viruses from sputum samples were significantly higher than those from nasopharyngeal swabs in adults using real-time multiplex RT-PCR. These findings suggest that sputum would benefit for the detection of respiratory viruses by nucleic acid amplification tests (NAATs) in patients who produce sputum. Further studies are needed to establish standardized RNA extraction methods from sputum samples. © 2014 Wiley Periodicals, Inc.

  5. Combined DECS Analysis and Next-Generation Sequencing Enable Efficient Detection of Novel Plant RNA Viruses

    Directory of Open Access Journals (Sweden)

    Hironobu Yanagisawa

    2016-03-01

    Full Text Available The presence of high molecular weight double-stranded RNA (dsRNA within plant cells is an indicator of infection with RNA viruses as these possess genomic or replicative dsRNA. DECS (dsRNA isolation, exhaustive amplification, cloning, and sequencing analysis has been shown to be capable of detecting unknown viruses. We postulated that a combination of DECS analysis and next-generation sequencing (NGS would improve detection efficiency and usability of the technique. Here, we describe a model case in which we efficiently detected the presumed genome sequence of Blueberry shoestring virus (BSSV, a member of the genus Sobemovirus, which has not so far been reported. dsRNAs were isolated from BSSV-infected blueberry plants using the dsRNA-binding protein, reverse-transcribed, amplified, and sequenced using NGS. A contig of 4,020 nucleotides (nt that shared similarities with sequences from other Sobemovirus species was obtained as a candidate of the BSSV genomic sequence. Reverse transcription (RT-PCR primer sets based on sequences from this contig enabled the detection of BSSV in all BSSV-infected plants tested but not in healthy controls. A recombinant protein encoded by the putative coat protein gene was bound by the BSSV-antibody, indicating that the candidate sequence was that of BSSV itself. Our results suggest that a combination of DECS analysis and NGS, designated here as “DECS-C,” is a powerful method for detecting novel plant viruses.

  6. Validation of a competitive ELISA and a virus neutralization test for the detection and confirmation of antibodies to Senecavirus A in swine sera.

    Science.gov (United States)

    Goolia, Melissa; Vannucci, Fabio; Yang, Ming; Patnayak, Devi; Babiuk, Shawn; Nfon, Charles K

    2017-03-01

    Senecavirus A (SVA; family Picornaviridae) is a nonenveloped, single-stranded RNA virus associated with idiopathic vesicular disease (IVD) in swine. SVA was detected in pigs with IVD in Brazil, United States, Canada, and China in 2015, triggering the need to develop and/or validate serologic assays for SVA. Our objective was to fully validate a previously developed competitive enzyme-linked immunosorbent assay (cELISA) as a screening test for antibodies to SVA. Additional objectives included the development and validation of a virus neutralization test (VNT) as a confirmatory test for SVA antibody detection, and the comparison of the cELISA, VNT, and an existing immunofluorescent antibody test (IFAT) for the detection of SVA antibodies in serial bleeds from SVA outbreaks. The diagnostic specificity and sensitivity were 98.2% (97.2-98.9%) and 96.9% (94.5-98.4%) for the cELISA, and 99.6% (99.0-99.9%) and 98.2% (95.8-99.4%) for the VNT, respectively. There was strong agreement among cELISA, VNT, and IFAT when compared based on kappa coefficient. Based on these performance characteristics, these tests are considered suitable for serologic detection of SVA in pigs.

  7. Ultrasensitive Detection of RNA and DNA Viruses Simultaneously Using Duplex UNDP-PCR Assay

    Science.gov (United States)

    Wang, Zengguo; Zhang, Xiujuan; Zhao, Xiaomin; Du, Qian; Chang, Lingling; Tong, Dewen

    2015-01-01

    Mixed infection of multiple viruses is common in modern intensive pig rearing. However, there are no methods available to detect DNA and RNA viruses in the same reaction system in preclinical level. In this study, we aimed to develop a duplex ultrasensitive nanoparticle DNA probe-based PCR assay (duplex UNDP-PCR) that was able to simultaneously detect DNA and RNA viruses in the same reaction system. PCV2 and TGEV are selected as representatives of the two different types of viruses. PCV2 DNA and TGEV RNA were simultaneously released from the serum sample by boiling with lysis buffer, then magnetic beads and gold nanoparticles coated with single and/or duplex specific probes for TGEV and PCV2 were added to form a sandwich-like complex with nucleic acids released from viruses. After magnetic separation, DNA barcodes specific for PCV2 and TGEV were eluted using DTT and characterized by specific PCR assay for specific DNA barcodes subsequently. The duplex UNDP-PCR showed similar sensitivity as that of single UNDP-PCR and was able to detect 20 copies each of PCV2 and TGEV in the serum, showing approximately 250-fold more sensitivity than conventional duplex PCR/RT-PCR assays. No cross-reaction was observed with other viruses. The positive detection rate of single MMPs- and duplex MMPs-based duplex UNDP-PCR was identical, with 29.6% for PCV2, 9.3% for TGEV and 3.7% for PCV2 and TGEV mixed infection. This duplex UNDP-PCR assay could detect TGEV (RNA virus) and PCV2 (DNA virus) from large-scale serum samples simultaneously without the need for DNA/RNA extraction, purification and reverse transcription of RNA, and showed a significantly increased positive detection rate for PCV2 (29%) and TGEV (11.7%) preclinical infection than conventional duplex PCR/RT-PCR. Therefore, the established duplex UNDP-PCR is a rapid and economical detection method, exhibiting high sensitivity, specificity and reproducibility. PMID:26544710

  8. Protocol for cost effective detection of cassava mosaic virus ...

    African Journals Online (AJOL)

    Early detection of cassava mosaic disease (CMD) is an extremely important step in containing the spread of the disease in Africa. Many nucleic acid based detection tools have been developed for CMD diagnosis but although these methods are specific and sensitive for their target DNA, they are not fast, cost effective, can't ...

  9. Simultaneous detection of four foodborne viruses in food samples using a one-step multiplex reverse transcription PCR.

    Science.gov (United States)

    Lee, Shin-Young; Kim, Mi-Ju; Kim, Hyun-Joong; Jeong, KwangCheol Casey; Kim, Hae-Yeong

    2017-11-15

    A one-step multiplex reverse transcription PCR (RT-PCR) method comprising six primer sets (for the detection of norovirus GI and GII, hepatitis A virus, rotavirus, and astrovirus) was developed to simultaneously detect four kinds of pathogenic viruses. The size of the PCR products for norovirus GI and GII, hepatitis A virus (VP3/VP1 and P2A regions), rotavirus, and astrovirus were 330, 164, 244, 198, 629, and 449 bp, respectively. The RT-PCR with the six primer sets showed specificity for the pathogenic viruses. The detection limit of developed multiplex RT-PCR, as evaluated using serially diluted viral RNAs, was comparable to that of one-step single RT-PCR. Also, this multiplex RT-PCR was evaluated using food samples such as water, oysters, lettuce, and vegetable product. These food samples were artificially spiked with four kinds of viruses in diverse combinations, and the spiked viruses in all food samples were detected successfully.

  10. Development of an improved RT-LAMP assay for detection of currently circulating rubella viruses.

    Science.gov (United States)

    Abo, H; Okamoto, K; Anraku, M; Otsuki, N; Sakata, M; Icenogle, J; Zheng, Q; Kurata, T; Kase, T; Komase, K; Takeda, M; Mori, Y

    2014-10-01

    Rubella virus is the causative agent of rubella. The symptoms are usually mild, and characterized by a maculopapular rash and fever. However, rubella infection in pregnant women sometimes can result in the birth of infants with congenital rubella syndrome (CRS). Global efforts have been made to reduce and eliminate CRS. Although a reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay for detection of rubella virus has been reported, the primers contained several mismatched nucleotides with the genomes of currently circulating rubella virus strains. In the present study, a new RT-LAMP assay was established. The detection limit of this assay was 100-1000PFU/reaction of viruses for all rubella genotypes, except for genotype 2C, which is not commonly found in the current era. Therefore, the new RT-LAMP assay can successfully detect all current rubella virus genotypes, and does not require sophisticated devices like TaqMan real-time PCR systems. This assay should be a useful assay for laboratory diagnosis of rubella and CRS. Copyright © 2014 Elsevier B.V. All rights reserved.

  11. Novel approach for detection of enteric viruses to enable syndrome surveillance of acute viral gastroenteritis.

    Science.gov (United States)

    Svraka, Sanela; van der Veer, Bas; Duizer, Erwin; Dekkers, Jojanneke; Koopmans, Marion; Vennema, Harry

    2009-06-01

    Acute gastroenteritis is one of the most common diseases worldwide, with viruses, particularly noroviruses, being the leading cause in developed countries. In The Netherlands, systematic surveillance of gastroenteritis outbreaks of suspected viral etiology was established by the National Institute for Public Health and the Environment in 1994. Since 2002, the total number of outbreaks reported has been increasing, and with that comes the need for sensitive assays that can be performed quickly. In addition, the diagnostic demand changed so that now the proportion of samples from hospitals is higher and there is a need for patient-based test results. In order to target the diagnosis of acute gastroenteritis, we reviewed our data on outbreaks of gastroenteritis and the prevalence of individual viruses to provide a priority list of viruses for which samples should be evaluated. Random primers were used to replace the separate specific primers for each virus used in the reverse transcription steps. The individual PCR assays were replaced by multiplex PCR assays. We employed a two-step method in which in the first step we screened for the most common causes of viral gastroenteritis, noroviruses of genogroup II and rotaviruses of group A, with equine arteritis virus used as the internal control. Subsequently, in the second step, two parallel PCR assays were developed for the detection of noroviruses of genogroup I and equine arteritis virus in one run and adenoviruses, sapoviruses, and astroviruses in the other run. The specificities of the assays were calculated to be 92.5% for the assay for noroviruses of genogroup I and 100% for the assays for all other viruses, the detection limits were equal for all viruses, and the turnaround time was reduced to 1 day compared to the at least 3 days required for the methods used previously. This approach allows the targeted, rapid, and cost-effective elucidation of the causes of acute gastroenteritis outbreaks.

  12. First detection of Toscana virus RNA from sand flies in the genus Phlebotomus (Diptera: Phlebotomidae) naturally infected in Morocco.

    Science.gov (United States)

    Es-Sette, N; Nourlil, J; Hamdi, S; Mellouki, F; Lemrani, M

    2012-11-01

    In total, 656 sand flies were collected in June 2008 from Louata, a locality of Sefrou province, Morocco. Testing was conducted for the presence of phlebovirus by nested reverse transcriptase polymerase chain reaction and sequencing. We detected Toscana virus in four pools of male Phlebotomus perniciosus. This virus belongs to the genotype B previously recognized in France and Spain. This is the first time that Toscana virus has been detected in Morocco.

  13. Isolamento do vírus rábico em morcego insetívoro, Nyctinomops macrotis, no Município de Diadema, SP (Brasil Isolation of rabies virus in an insectivorous bat Nyctinomops macrotis, in Southeastern Brazil

    Directory of Open Access Journals (Sweden)

    Estevão C. Passos

    1998-02-01

    Full Text Available Foi realizado o isolamento do vírus rábico em morcego insetívoro Nyctinomops macrotis capturado próximo à represa Billings e à mata Atlântica, no Município de Diadema, SP (Brasil. A pesquisa do antígeno rábico no tecido cerebral do morcego apresentou resultado positivo na reação de imunofluorescência direta. O isolamento do vírus rábico no tecido cerebral e nas glândulas salivares do morcego foi obtido através da inoculação intracerebral em camundongos. O Município de Diadema não apresentava casos de raiva animal desde 1982, sendo este o primeiro relato da presença do vírus rábico em morcego insetívoro.The rabies virus was isolated from an insectivorous bat, Nyctinomops macrotis, trapped in Diadema, SP, Brazil, in a public building near a water supply reservoir. Fluorescent antibodies against rabies virus were detected in cerebral tissue and the viral isolation was made after the inoculation of cerebral tissue and salivary gland suspension in mice. There have been no recorded cases of animal rabies in Diadema since 1982, and this is the first isolation of the rabies virus in an insectivorous bat in the city.

  14. Molecular and clinical epidemiological surveillance of dengue virus in Paraíba, Northeast Brazil

    Directory of Open Access Journals (Sweden)

    Isabel Cristina Guerra-Gomes

    Full Text Available ABSTRACT INTRODUCTION: Despite being the most prevalent arboviral disease worldwide, dengue has been neglected lately. However, recent epidemics of arboviruses such as Zika and chikungunya in locations throughout the world have alerted health authorities to these diseases. This study evaluated the incidence pattern of dengue, its clinical characteristics, and co-circulation of serotypes from 2007 to 2015 in Paraiba State, Northeast Brazil. METHODS: Data on dengue cases from 2007 to 2015 were extracted from clinical reports of the National System for Notifiable Diseases [Sistema Nacional de Agravos de Notificação (SINAN] of Brazil provided by the Paraiba Health Department. Reverse transcription polymerase chain reaction (RT-PCR assays for dengue serotypes were carried out on plasma samples obtained from patients with suspected dengue. The data were analysed using descriptive statistics. RESULTS: According to clinical features, dengue fever [n = 39,083 (70.2%] and dengue without warning signs [n = 15,365 (27.7%] were the most common classifications of dengue. On RT-PCR, DENV 1 was the most commonly identified serotype (80.5% in all years studied. Co-circulation of all four DENV serotypes was observed in 2013 and 2014. Furthermore, we observed an increase in dengue notifications in 2015, possibly due to the rise of Zika and chikungunya. CONCLUSIONS: Our findings support the hypothesis that co-circulation of the four DENV serotypes may be a reason for the increased prevalence of severe forms of dengue in the years studied. This study may contribute to directing research, health policy, and financial resources toward reducing poorly controlled epidemic diseases.

  15. Detection of viruses directly from the fresh leaves of a Phalaenopsis orchid using a microfluidic system.

    Science.gov (United States)

    Chang, Wen-Hsin; Yang, Sung-Yi; Lin, Chih-Lin; Wang, Chih-Hung; Li, Ping-Chen; Chen, Tzong-Yueh; Jan, Fuh-Jyh; Lee, Gwo-Bin

    2013-11-01

    Early detection of pathogens is crucial for the effective surveillance of diseases. Many efforts have been made to explore methods which can detect these pathogens within a short period of time without requiring a tedious protocol. However, these developed methods have disadvantages such as they are relatively time-consuming or require specialized laboratory facilities. In this work, we have developed an integrated microfluidic system for rapid and automatic detection of viruses by direct analysis from fresh Phalaenopsis orchid leaves. The entire protocol, including ribonucleic acid (RNA) purification, reverse transcription loop-mediated-isothermal-amplification (RT-LAMP) and optical detection by measuring changes in turbidity was performed on a single chip. This is the first time that an integrated microfluidic system for the detection of viruses infecting the Phalaenopsis orchid has been demonstrated. The sensitivity of the developed system was also explored in this study to validate its performance. In this study, the authors report the development of an integrated microfluidic system for rapid and automatic detection of viruses by direct analysis of fresh Phalaenopsis orch