Sample records for amplicor hiv-i monitortju

  1. Evaluation of the Amplicor Chlamydia trachomatis test versus culture in genital samples in various prevalence populations.

    de Barbeyrac, B.; Pellet, I; Dutilh, B.; Dumon, B.; Géniaux, M; Bébéar, C.


    OBJECTIVE--To evaluate a newly developed polymerase chain reaction (PCR) assay, Amplicor C trachomatis for the detection of C trachomatis in genital samples using cell culture for comparison. SUBJECTS--501 patients (431 women and 70 men) attending an STD clinic in Hôpital Pellegrin (high-risk population) and gynaecological clinics (low-risk population) in Bordeaux, France. METHODS--The genital samples (cervical and urethral) were tested for the presence of C trachomatis using the Amplicor tes...

  2. Evaluation of automated COBAS AMPLICOR PCR system for detection of several infectious agents and its impact on laboratory management.

    Jungkind, D; Direnzo, S; Beavis, K G; Silverman, N S


    We evaluated the COBAS AMPLICOR (CA) PCR system (Roche Diagnostic Systems) designed for automated PCR amplification and detection of nucleic acids from infectious agents in clinical samples. The Roche AMPLICOR microwell plate (MWP) PCR was the reference method. CA amplifies target nucleic acid, captures the biotinylated amplification products by using magnetic particles coated with specific oligonucleotide probes, and detects the bound products colorimetrically. For Mycobacterium tuberculosis...

  3. Comparison of COBAS AMPLICOR Neisseria gonorrhoeae PCR, including confirmation with N. gonorrhoeae-specific 16S rRNA PCR, with traditional culture

    Luijt, D.S.; Bos, P.A.; van Zwet, A.A.; Voorst-Vader, P.C.; Schirm, J.


    : A total of 3,023 clinical specimens were tested for Neisseria gonorrhoeae by using COBAS AMPLICOR (CA) PCR and confirmation of positives by N. gonorrhoeae-specific 16S rRNA PCR. The sensitivity of CA plus 16S rRNA PCR was 98.8%, compared to 68.2% for culture. Confirmation of CA positives increased

  4. Comparison of COBAS AMPLICOR Neissefia gonorrhoeae PCR, including confirmation with N-gonorrhoeae-specific 16S rRNA PCR, with traditional culture

    Luijt, DS; Bos, PAJ; van Zwet, AA; Vader, PCV; Schirm, J


    A total of 3,023 clinical specimens were tested for Neisseria gonorrhoeae by using COBAS AMPLICOR (CA) PCR and confirmation of positives by N. gonorrhoeae-specific 16S rRNA PCR. The sensitivity of CA plus 16S rRNA PCR was 98.8%, compared to 68.2% for culture. Confirmation of CA positives increased t

  5. Comparison of real-time polymerase chain reaction with the COBAS Amplicor test for quantitation of hepatitis B virus DNA in serum samples

    Ming Shi; Yong Zhang; Ying-Hua Zhu; Jing Zhang; Wei-Jia Xu


    AIM: To compare the clinical performance of a real-time PCR assay with the COBAS Amplicor Hepatitis B Virus (HBV) Monitor test for quantitation of HBV DNA in serum samples. METHODS: The reference sera of the Chinese National Institute for the Control of Pharmaceutical and Biological Products and the National Center for Clinical Laboratories of China, and 158 clinical serum samples were used in this study. The linearity, accuracy, reproducibility, assay time, and costs of the real-time PCR were evaluated and compared with those of the Cobas Amplicor test. RESULTS: The intra-assay and inter-assay variations of the real-time PCR ranged from 0.3% to 3.8% and 1.4% to 8.1%, respectively. The HBV DNA levels measured by the real-time PCR correlated very well with those obtained with the COBAS Amplicor test (r = 0.948). The real-time PCR HBV DNA kit was much cheaper and had a wider dynamic range. CONCLUSION: The real-time PCR assay is an excellent tool for monitoring of HBV DNA levels in patients with chronic hepatitis B.

  6. Comparison of the Abbott m2000 HIV-1 Real-Time and Roche AMPLICOR Monitor v1.5 HIV-1 assays on plasma specimens from Rakai, Uganda

    Ssebugenyi, I.; Kizza, A; Mpoza, B; Aluma, G.; Boaz, I.; Newell, K.; Laeyendecker, O; Shott, J P; Serwadda, D; Reynolds, S.J.


    The need for viral load (VL) monitoring of HIV patients receiving antiretroviral therapy (ART) in resource-limited settings (RLS) has become apparent with studies showing the limitations of immunological monitoring. We compared the Abbott m2000 Real-Time (Abbott) HIV-1 assay with the Roche AMPLICOR Monitor v1.5 (Roche) HIV-1 assay over a range of VL concentrations. Three hundred and eleven plasma samples were tested, including 164 samples from patients on ART ≥ six months and 147 from ART-naï...

  7. Comparison of the Sensitivities of the Version 1.5 and Version 1.0 Ultrasensitive Roche AMPLICOR HIV-1 MONITOR Kits at Low Concentrations of Human Immunodeficiency Virus RNA

    Brambilla, Donald J.; Jennings, Cheryl; Morack, Ralph; Granger, Suzanne; Bremer, James W.


    The sensitivities of the version 1.5 and 1.0 Roche UltraSensitive AMPLICOR HIV-1 MONITOR tests were compared using panels of coded samples of subtype B human immunodeficiency virus type 1 spiked into plasma at predetermined concentrations. Results indicate that the version 1.5 kit is more sensitive than the version 1.0 kit.

  8. Computer tools in the discovery of HIV-I integrase inhibitors

    Liao, Chenzhong; Nicklaus, Marc C.


    Computer-aided drug design (CADD) methodologies have made great advances and contributed significantly to the discovery and/or optimization of many clinically used drugs in recent years. CADD tools have likewise been applied to the discovery of inhibitors of HIV-I integrase, a difficult and worthwhile target for the development of efficient anti-HIV drugs. This article reviews the application of CADD tools, including pharmacophore search, quantitative structure–activity relationships, model b...

  9. Comparison of the Abbott m2000 RealTime CT Assay and the Cepheid GeneXpert CT/NG Assay to the Roche Amplicor CT Assay for Detection of Chlamydia trachomatis in Ocular Samples from Tanzania

    Dize, Laura; West, Sheila; Williams, James A; Van Der Pol, Barbara; Quinn, Thomas C.; Gaydos, Charlotte A.


    The GeneXpert CT/NG assay (GeneXpert) and the Abbott m2000 RealTime CT (m2000) assay were compared to Amplicor for detecting ocular Chlamydia trachomatis. Discordant specimens were tested by the Aptima CT assay. The m2000 assay sensitivity was 100% (95% confidence interval [CI], 90% to 100%), and specificity was 98.46% (95% CI, 95.2% to 99.2%); GeneXpert sensitivity was 100% (95% CI, 90% to 100%), and specificity was 100% (95% CI, 98.1% to 100%). The m2000 and GeneXpert assays appear to perfo...

  10. Evaluation of the NucliSens EasyQ v2.0 assay in comparison with the Roche Amplicor v1.5 and the Roche CAP/CTM HIV-1 Test v2.0 in quantification of C-clade HIV-1 in plasma.

    Maximilian Muenchhoff

    Full Text Available Human immunodeficiency virus type 1 (HIV-1 genetic diversity poses a challenge to reliable viral load monitoring. Discrepancies between different testing platforms have been observed, especially for non-clade-B virus. Therefore we compare, in antiretroviral therapy (ART-naïve South African subjects predominantly infected with HIV-1 clade-C, three commercially available assays: the COBAS AmpliPrep/COBAS TaqMan HIV-1 Test version 2.0 by Roche (CAP/CTM v2.0, the BioMérieux NucliSens Version 2.0 Easy Q/Easy Mag (NucliSens v2.0 and the Roche COBAS Amplicor HIV-1 Monitor Test Version 1.5 (Amplicor v1.5. Strong linear correlation was observed and Bland-Altman analyses showed overall good agreement between the assays with mean viral load differences of 0.078 log cp/ml (NucliSens v2.0 - Amplicor v1.5, 0.260 log cp/ml (CAP/CTM v2.0 - Amplicor v1.5 and 0.164 log cp/ml (CAP/CTM v2.0 - NucliSens v2.0, indicating lower mean viral load results for the Amplicor v1.5 and higher mean readings for the CAP/CTM v2.0. Consistent with observations following previous comparisons of CAP/CTM v2.0 versus Amplicor v1.5, the CAP/CTM v2.0 assay detected low-level viremia (median 65 cp/ml in more than one-third of those in whom viremia had been undetectable (<20 cp/ml in assays using the NucliSens platform. These levels of viremia are of uncertain clinical significance but may be of importance in early detection of ART resistance in those on treatment. Overall the three assays showed good comparability of results but with consistent, albeit relatively small, discrepancies for HIV-1 clade-C samples, especially in the low-viremic range that should be taken into account when interpreting viral load data.

  11. A novel laccase with inhibitory activity towards HIV-I reverse transcriptase and antiproliferative effects on tumor cells from the fermentation broth of mushroom Pleurotus cornucopiae.

    Wu, Xiangli; Huang, Chenyang; Chen, Qiang; Wang, Hexiang; Zhang, Jinxia


    A novel laccase with a molecular mass of 67 kDa was isolated from the fermentation broth of Pleurotus cornucopiae through ion exchange chromatography and gel filtration. The optimal pH and temperature for the laccase was pH 4.2 and 30°C, respectively. The laccase activity was remarkably inhibited by Fe(3+) and Hg(2+) , while it was stimulated by Cu(2+) and Pb(2+) . It inhibited proliferation of the hepatoma cells HepG2 and the breast cancer cells MCF-7, and the activity of HIV-I reverse transcriptase with IC50 values of 3.9, 7.6 and 3.7 μM, respectively. PMID:24136666

  12. Spatial Genetic Structure of Two HIV-I-resistant Polymorphisms (CCR2-64Ⅰand SDF1-3'A) Alleles in Population of Shandong Province, China


    Objective To explore the spatial genetic structure of two HIV-I-resistant polymorphisms (CCR2-64Ⅰand SDF1-3'A) alleles in the population of Shandong Province, China. Methods Using the techniques of spatial stratified sampling and spatial statistics, the spatial genetic structure of the locus (CCR2-64Ⅰand SDF1-3'A), which was shown to be important co-receptor for HIV infection, was quantified from the populations of 36 sampled counties of Shandong Province, and a total of 3147 and 3172 samples were taken for testing CCR2-64I and SDF1-3'A respectively from individuals without known history of HIV-I infection and AIDS symptoms. Results There were significantly spatial genetic structures of the two alleles at different spatial distance classes on the scale of populations, but on the scale of individuals, no spatial structure was found in either the whole area of Shandong Province or the area of each sampled county. Although the change of frequencies of the two alleles with geographic locations in Shandong Province both showed gradual increase trends, their changing directions were inverse. The frequency of CCR2-64I allele gradually increased from the southwest to the northeast, while the frequency of SDF1-3'A allele gradually increased from the northeast to the southwest. However the RH to AIDS of combined types of their different genotypes did not represent obvious geographic diversity on the whole area of the Province. Conclusion The frequency of allele usually has some spatial genetic structures or spatial autocorrelation with different spatial distance classes, but the genotypes of individuals have random distribution in the same geographic area. Evaluating spatial distribution of the genetic susceptibility of HIV (AIDS) to CCR2-64I and SDF1-3'A alleles, should focus on the frequencies of combined genotypes of CCR2 and SDF1 based on the two-locus genotypes of each individual rather than the frequencies of CCR2-64I and SDF1-3'A alleles.

  13. Vertikal smitte med hiv i Danmark

    Kvinesdal, Birgit Bak; Valerius, Niels Henrik; Herlin, Troels;


    INTRODUCTION: Vertical transmission of HIV can be reduced if the pregnant woman and new born child receive antiretroviral treatment. Delivery by caesarean section and avoidance of breast feeding further reduce vertical transmission. The aim of this study was to describe the treatment of HIV......-positive pregnant women in Denmark and the risk of vertical transmission. MATERIAL AND METHODS: We retrospectively describe the risk of vertical transmission of HIV among HIV-positive women giving birth in Denmark during the period, mid-1994 to February 2000. RESULTS: Fifty children were born. One mother gave birth...... mothers was the HIV-infection known until the time of delivery or later. Transmission of HIV did not occur in the 34 mother-child pairs who received antepartum and intrapartum antiretroviral treatment, who had a caesarean delivery, who did not breast-feed, and whose children were given postpartum...

  14. Rapid diagnosis of enterovirus infection by a new one-step reverse transcription-PCR assay.

    Kessler, H H; Santner, B; Rabenau, H; Berger, A; Vince, A; Lewinski, C; B. Weber; Pierer, K.; Stuenzner, D; Marth, E; Doerr, H W


    The AMPLICOR Enterovirus Test was evaluated with 103 cerebrospinal fluid (CSF) specimens. Twenty-seven CSF specimens were culture positive. With the AMPLICOR test, enterovirus RNA was detected in 34 specimens. Compared with culture, the AMPLICOR test gave a sensitivity of 96.3% and a specificity of 100%. The sensitivity of culture was 79.4% in comparison with the AMPLICOR test.

  15. HIV-i võib nakatunud olla iga sajas täiskasvanud eestlane / Marika Raiski

    Raiski, Marika


    Vt. ka Nädaline 27. sept., lk. 7. Vastavalt Maailma Terviseorganisatsiooni (WHO) ja UNAIDS-i hinnangule on Eestis hulgaliselt registreerimata HIV-juhte ning tegelikult võib HIV-nakatunuid olla juba iga sajas täiskasvanud Eesti elanik

  16. Vad påverkar HIV i Sydafrika? : En teoretisk och empirisk analys av Sydafrikas provinser 2008

    Skogum, Anna; Sjögren, Anna


    What affects Hiv in South Africa's different provinces? This thesis examines if education, unemployment rate, GDP per capita, and the literacy rate has any relationship with the Hiv prevalence. This is analyzed using theory and data in the form of a simple microeconomic model and an econometric regression analysis based on cross sectional data of the provinces of South Africa. The regression analysis shows that unemployment rate and education have significant effects of the Hiv prevalence in ...

  17. Comparison of two commercial assays for detection of human papillomavirus (HPV) in cervical scrape specimens: validation of the Roche AMPLICOR HPV test as a means to screen for HPV genotypes associated with a higher risk of cervical disorders.

    Ham, M.A. van; Bakkers, J.M.J.E.; Harbers, G.; Quint, W.G.V.; Massuger, L.F.A.G.; Melchers, W.J.G.


    Certain high-risk (HR) human papillomavirus (HPV) types are a necessary cause for the development of cervical disorders. Women with persistent HR HPV infections have an increased risk of developing high-grade cervical lesions, compared with those who have no or low-risk HPV infections. Therefore, im

  18. Potent Functional Antibody Responses Elicited by HIV-I DNA Priming and Boosting with Heterologous HIV-1 Recombinant MVA in Healthy Tanzanian Adults

    Agricola Joachim; Charlotta Nilsson; Said Aboud; Muhammad Bakari; Lyamuya, Eligius F; Merlin L Robb; Marovich, Mary A.; Patricia Earl; Bernard Moss; Christina Ochsenbauer; Britta Wahren; Fred Mhalu; Eric Sandström; Gunnel Biberfeld; Guido Ferrari


    Vaccine-induced HIV antibodies were evaluated in serum samples collected from healthy Tanzanian volunteers participating in a phase I/II placebo-controlled double blind trial using multi-clade, multigene HIV-DNA priming and recombinant modified vaccinia Ankara (HIV-MVA) virus boosting (HIVIS03). The HIV-DNA vaccine contained plasmids expressing HIV-1 gp160 subtypes A, B, C, Rev B, Gag A, B and RTmut B, and the recombinant HIV-MVA boost expressed CRF01_AE HIV-1 Env subtype E and Gag-Pol subtyp...

  19. The epidemiology of HIV seropositive malaria infected pregnant women in Akure Metropolis, Southwestern Nigeria

    Ajibade Kwashie Ako-Nai


    Full Text Available Background: HIV increases the risks of malaria in pregnant women, while maternal human immunodeficiency virus (HIV viral load also facilitates perinatal transmission to neonates. Malaria and HIV coinfection has been shown to exacerbate adverse pregnancy complications. Our study was designed to determine the HIV prevalence of pregnant women at an antenatal clinic in Akure in southwestern Nigeria, investigate the relationship between dual HIV and malaria infection and HIV viral load and CD4+ T cell counts. The study also estimated the risks of adverse pregnancy outcomes in a selected cohort of 74 pregnant women. Materials and Methods: We evaluated the HIV serostatus of 3,225 pregnant women, who attended the antenatal clinic between August 2012 and April 2013. A cohort of 74 pregnant women was selected for the investigation of the relationship between coinfection of HIV and malaria and HIV viral load and CD4+ cell counts. Their HIV status was determined during three trimesters of pregnancy by both HIV-1/2 strips and confirmatory enzyme-linked immunosorbent assay (ELISA method. Malaria parasitemia was determined by Giemsa-stained thin and thick blood smears. CD4 cell count was by flow cytometry using the CyFlow Counter (Partec, Germany. Viral load estimated by Amplicor HIV-I monitor assay. Results: We found 3.53% prevalence of HIV serostatus among the 3,225 pregnant women who were screened. Forty-four of the 74 subjects were HIV positive and 30 were HIV negative controls. The results show HIV infection among the pregnant women reduced the CD4 cells from a mean of 750 cells/ml for HIV negative women to a mean of 363 cells/ml for HIV seropositive women. Additionally the presence of malaria more than doubled the HIV viral load from a mean of 7,270 ribonucleic acid (RNA copies/ml for HIV positive women without malaria to 15,148 RNA copies/ml for HIV positive women with malaria. Conclusion: In this study, HIV infection significantly increased risk of

  20. Comparison of in-house and commercial sample preparation and PCR amplification systems for detection of human immunodeficiency virus type 1 DNA in blood samples from Tanzanian adults.

    Lyamuya, E; Bredberg-Rådén, U; Albert, J.; Grankvist, O; Msangi, V; Kagoma, C.; Mhalu, F; Biberfeld, G


    This study compared the performance of several in-house nested PCR systems and the Amplicor human immunodeficiency virus type 1 (HIV-1) PCR kit in the detection of HIV-1 DNA in Tanzanian samples prepared by two different methods. All six of the in-house primer sets evaluated had a higher sensitivity for HIV DNA detection in samples prepared by the Amplicor PCR sample preparation method than in those prepared by the Ficoll-Isopaque (FIP) density gradient centrifugation method. A sensitivity of...

  1. Correlation of hepatitis C RNA and serum alanine aminotransferase in hepatitis B and C seronegative healthy blood donors

    Ali Natasha


    Full Text Available Introduction: Historically, serum alanine transaminase (ALT has been used as a surrogate marker in the detection of hepatitis viruses in blood donors. With the availability of newer sensitive technologies for the detection of seroconversion, the value of ALT becomes questionable but continues to be used for this purpose with subsequent discarding of ALT elevated blood units. Objective: The present study aims to evaluate the significance and cost effectiveness of ALT as a surrogate marker for hepatitis C virus infection in healthy asymptomatic blood donors who were serologically negative. Materials and Methods: The study was conducted at clinical laboratory of a tertiary care hospital for a period of one year from November 2006 to October 2007. All donors were screened serologically for hepatitis B, C and HIV I and II, syphilis and malaria and those tested positive were excluded from further evaluation. Gender-wise reference ranges and minimal and markedly raised results for ALT (described respectively as one and two folds increase above reference range were defined and, accordingly, donors were grouped into three. Two hundred seronegative blood donors were randomly selected from all three groups of ALT results and tested for hepatitis C nucleic acid through Amplicor; HCV RNA test. The cost of discarding an ALT -only elevated blood unit was also assessed. During the study period, 25117 subjects donated blood. Eight hundred and Results: seventy two donors (3.4% were positive for one or more serological tests. ALT of all donors ranged from 0-1501 U/L (Mean ± SD; 33.4 ± 25.45U/L. The donors seronegative for all disease markers were 24245 (96.6%. Of these, 21164 (87.2% donors had their ALT within reference range while 2874 (11.8% and 207 (0.8% of donors had minimal and markedly elevated results. Thus, 621 blood bags (red cells, platelets and plasma costing $ 39200.0 were discarded based on ALT results alone. Of 200 seronegative donors evaluated

  2. Multicenter evaluation of the new Abbott Realtime assays for quantitative detection of human immunodeficiency virus type 1 and hepatitis C virus RNA

    M. Schutten (Martin); D. Peters (D.); N. Back (Nicole); A.W. van den Beld (Annewieke); B. Beuselinck (B.); V. Foulongne (V.); A.M. Geretti (Anna Maria); L. Pandiani (L.); M. Tiemann; H.G.M. Niesters (Bert)


    textabstractThe analytical performances of the new Abbott RealTime hepatitis C virus (HCV) and human immunodeficiency virus type 1 viral load assays were compared at nine laboratories with different competitor assays. These included the Abbott LcX, Bayer Versant bDNA, Roche COBAS Amplicor, and Roche

  3. Multicenter evaluation of the new Abbott RealTime assays for quantitative detection of human immunodeficiency virus type 1 and hepatitis C virus RNA

    Schutten, M; Peters, D; Back, N K T; Beld, M; Beuselinck, K; Foulongne, V; Geretti, A-M; Pandiani, L; Tiemann, C; Niesters, H G M


    The analytical performances of the new Abbott RealTime hepatitis C virus (HCV) and human immunodeficiency virus type 1 viral load assays were compared at nine laboratories with different competitor assays. These included the Abbott LcX, Bayer Versant bDNA, Roche COBAS Amplicor, and Roche COBAS TaqMa

  4. Quantitation of HIV-1 RNA in breast milk by real time PCR

    Becquart, Pierre; Foulongne, Vincent; Willumsen, J.; Rouzioux, C; Segondy, Michel; Van De Perre, Philippe


    HIV-1 RNA in breast milk is a strong predictor of HIV-1 transmission through breastfeeding. In the present report, breast milk samples from HIV-1 uninfected donors were spiked with dilution of quantified culture supernatant from HIV-1(NDK) infected PBMC. Two RNA extraction techniques based on silica extraction, Nuclisens (R) (BioMerieux) and Triazol (Qiagen), two techniques based on guanidine thiocynanate/chloroforme extraction, TRIzol (Life Technologic) and Amplicor HIV-1 Monitor (TM) (Roche...

  5. Clinical characterization of a competitive PCR assay for quantitative testing of hepatitis C virus.

    Miskovsky, E P; Carrella, A V; Gutekunst, K; Sun, C. A.; Quinn, T C; Thomas, D L


    Rational clinical application of quantitative assessments of hepatitis C virus (HCV) RNA depends on an understanding of factors affecting the assay and its intrinsic variability. The effects of three types of blood collection tubes, two storage temperatures, five processing times, and two laboratories on a commercially available quantitative reverse transcriptase PCR assay (AMPLICOR HCV MONITOR) were evaluated. HCV RNA concentrations were assessed in 356 specimens representing 178 aliquots fr...

  6. Detection of intrahepatic hepatitis B virus DNA and correlation with hepatic necroinflammation and fibrosis

    Wong, Danny Ka-Ho; Yuen, Man-Fung; Tse, Eric; Yuan, Hejun; Sum, Simon Siu-Man; Hui, Chee-Kin; Lai, Ching-Lung


    Assessment of intrahepatic hepatitis B virus (HBV) DNA levels in patients with chronic hepatitis B is important in understanding the natural history of the disease and designing antiviral therapy regimens. However, there is no standardized method for the measurement of intrahepatic HBV DNA levels. We describe a convenient novel method for the measurement of intrahepatic HBV DNA levels based on a modified COBAS Amplicor HBV Monitor test for HBV DNA measurement and real-time PCR β-actin gene de...

  7. External Quality Assessment Program for Chlamydia trachomatis Diagnostic Testing by Nucleic Acid Amplification Assays

    Land, Sally; Tabrizi, Sepehr; Gust, Anthony; Johnson, Elizabeth; Garland, Susan; Dax, Elizabeth M.


    We report the results from 57 Australian diagnostic laboratories testing two external quality assessment panels using either the Roche Amplicor Chlamydia trachomatis test (R-PCR) or the Abbott LCx Chlamydia trachomatis assay (A-ligase chain reaction [LCR]). Panel samples were either normal urine spiked with Chlamydia trachomatis antigen or clinical urine specimens. There was no significant difference between laboratories or between assays in detection of C. trachomatis-positive clinical sampl...

  8. Comparison of Two Measures of Human Immunodeficiency Virus (HIV) Type 1 Load in HIV Risk Groups

    Lyles, Cynthia M.; Vlahov, David; Farzadegan, Homayoon; Astemborski, Jacquie; Margolick, Joseph B.; Masters, Beth A.; Schroeder, Jennifer; Quinn, Thomas C


    Levels of viral burden were compared across risk group and gender populations among 485 human immunodeficiency virus type 1 (HIV-1)-infected participants consisting of 190 male injection drug users (IDUs), 92 female IDUs, and 203 homosexual men. Viral burden was quantified by a microculture technique to determine cell-associated infectious units per 106 peripheral blood mononuclear cells (IUPM) and by reverse transcriptase PCR (Amplicor) to determine plasma HIV RNA levels. Adjusting for CD4+ ...

  9. EUROarray human papillomavirus (HPV) assay is highly concordant with other commercial assays for detection of high-risk HPV genotypes in women with high grade cervical abnormalities.

    Cornall, A M; Poljak, M; Garland, S M; Phillips, S; Machalek, D A; Tan, J H; Quinn, M A; Tabrizi, S N


    The purpose of this study was to evaluate the performance of the EUROIMMUN EUROArray HPV genotyping assay against the Roche Cobas 4800, Roche HPV Amplicor, Roche Linear Array and Qiagen Hybrid Capture 2 assays in the detection of high-risk HPV (HR-HPV) from liquid based cervical cytology samples collected from women undergoing follow-up for abnormal cervical cytology results. Cervical specimens from 404 women undergoing management of high-grade cytological abnormality were evaluated by EUROarray HPV for detection of HR-HPV genotypes and prediction of histologically-confirmed cervical intraepithelial neoplasia grade 2 or higher (≥CIN2). The results were compared to Hybrid Capture 2, Cobas 4800 HPV, Amplicor and Linear Array HPV. Positivity for 14 HR-HPV types was 80.0 % for EUROarray (95 % CI; 75.7-83.8 %). Agreement (κ, 95 % CI) between the EUROarray and other HPV tests for detection of HR-HPV was good to very good [Hybrid Capture κ = 0.62 (0.54-0.71); Cobas κ = 0.81 (0.74-0.88); Amplicor κ = 0.68 (0.60-0.77); Linear Array κ = 0.77 (0.70-0.85)]. For detection of HR-HPV, agreement with EUROarray was 87.90 % (Hybrid Capture), 93.58 % (Cobas), 92.84 % (Amplicor) and 92.59 % (Linear Array). Detection of HR-HPV was not significantly different between EUROarray and any other test (p < 0.001). EUROarray was concordant with other assays evaluated for detection of high-risk HPV and showed sensitivity and specificity for detection of ≥ CIN2 of 86 % and 71 %, respectively. PMID:27048314

  10. Comparison of Two Amplification Technologies for Detection and Quantitation of Human Immunodeficiency Virus Type 1 RNA in the Female Genital Tract

    Bremer, James; Nowicki, Marek; Beckner, Suzanne; Brambilla, Donald; Cronin, Mike; Herman, Steven; Kovacs, Andrea; Reichelderfer, Patricia


    Human immunodeficiency virus type 1 (HIV-1) RNA levels in female genital tract and peripheral blood samples were compared using two commercial amplification technologies: the Roche AMPLICOR HIV-1 MONITOR test and either the Organon Teknika nucleic acid sequence-based amplification (NASBA-QT) assay or the NucliSens assay. Estimates of HIV-1 RNA copy number were derived from internal kit standards and analyzed unadjusted and adjusted to a common set of external standards. We found a discordance...

  11. Aids ähvardab Venemaa, Hiina ja India tulevikku / Neeme Raud

    Raud, Neeme, 1969-


    Ameerika analüütik Nicholas Eberstadt prognoosib, et Hiina, India ja Venemaa võivad järgmise veerandsaja jooksul seista silmitsi suurema arvu HIV-i nakatanute ja aidsi surnutega kui kogu maailm seni tervikuna

  12. Single-step PCR in molecular diagnosis of hepatitis C virus infection.

    Farma, E; E. Boeri; Bettini, P.; Repetto, C M; McDermott, J.; Lillo, F B; Varnier, O E


    The diagnostic utility of two PCR systems and three PCR detection methods for hepatitis C virus (HCV) RNA was evaluated in serum samples. A nested PCR was considered the reference assay and was compared with two single-step PCR methods: the first is based on the detection of PCR products by liquid hybridization with a 32P-end-labeled probe, and the second is the Roche Amplicor colorimetric assay using microwell plate hybridization with a specific nucleic acid probe. Using the Pelicheck HCV RN...

  13. Multicenter Evaluation of the New Abbott RealTime Assays for Quantitative Detection of Human Immunodeficiency Virus Type 1 and Hepatitis C Virus RNA▿

    Schutten, M.; Peters, D.; Back, N.K.T.; Beld, M; Beuselinck, K.; Foulongne, V.; Geretti, A.-M.; Pandiani, L.; Tiemann, C; Niesters, H.G.M.


    The analytical performances of the new Abbott RealTime hepatitis C virus (HCV) and human immunodeficiency virus type 1 viral load assays were compared at nine laboratories with different competitor assays. These included the Abbott LcX, Bayer Versant bDNA, Roche COBAS Amplicor, and Roche COBAS TaqMan assays. Two different protocols used during the testing period with and without a pre-m1000 RNA isolation spin were compared. The difference proved to be nonsignificant. A uracil-N-glycosylase (U...

  14. A comprehensive serological and supplemental evaluation of hepatitis B “seroyield” blood donors: A cross-sectional study from a tertiary healthcare center in India

    Prashant Pandey; Aseem K Tiwari; Dara, Ravi C.; Geet Aggarwal; Ganesh Rawat; Vimarsh Raina


    Background: The present study addressed the interesting findings of supplemental evaluation of hepatitis B "seroyield" donors. Materials and Methods: Each blood donor sample was tested for anti-human immunodeficiency virus type I (HIV-I)/HIV type II (HIV-II), HBsAg, and anti-hepatitis C virus (HCV) antibody by enhanced chemiluminescence method and subjected to individual donor-nucleic acid testing (NAT) for HIV-I, hepatitis B virus (HBV), and HCV. NAT test was performed using the eSAS system,...

  15. Systematic review of the performance of HIV viral load technologies on plasma samples.

    Kimberly A Sollis

    Full Text Available BACKGROUND: Viral load (VL monitoring is the standard of care in developing country settings for detecting HIV treatment failure. Since 2010 the World Health Organization has recommended a phase-in approach to VL monitoring in resource-limited settings. We conducted a systematic review of the accuracy and precision of HIV VL technologies for treatment monitoring. METHODS AND FINDINGS: A search of Medline and Embase was conducted for studies evaluating the accuracy or reproducibility of commercially available HIV VL assays. 37 studies were included for review including evaluations of the Amplicor Monitor HIV-1 v1.5 (n = 25, Cobas TaqMan v2.0 (n = 11, Abbott RealTime HIV-1 (n = 23, Versant HIV-1 RNA bDNA 3.0 (n = 15, Versant HIV-1 RNA kPCR 1.0 (n = 2, ExaVir Load v3 (n = 2, and NucliSens EasyQ v2.0 (n = 1. All currently available HIV VL assays are of sufficient sensitivity to detect plasma virus levels at a lower detection limit of 1,000 copies/mL. Bias data comparing the Abbott RealTime HIV-1, TaqMan v2.0 to the Amplicor Monitor v1.5 showed a tendency of the Abbott RealTime HIV-1 to under-estimate results while the TaqMan v2.0 overestimated VL counts. Compared to the Amplicor Monitor v1.5, 2-26% and 9-70% of results from the Versant bDNA 3.0 and Abbott RealTime HIV-1 differed by greater than 0.5log10. The average intra and inter-assay variation of the Abbott RealTime HIV-1 were 2.95% (range 2.0-5.1% and 5.44% (range 1.17-30.00% across the range of VL counts (2log10-7log10. CONCLUSIONS: This review found that all currently available HIV VL assays are of sufficient sensitivity to detect plasma VL of 1,000 copies/mL as a threshold to initiate investigations of treatment adherence or possible treatment failure. Sources of variability between VL assays include differences in technology platform, plasma input volume, and ability to detect HIV-1 subtypes. Monitoring of individual patients should be performed on the same

  16. CMV DNA e MRNA Pp67 nel monitoraggio del paziente sottoposto a trapianto di midollo osseo

    Maria Bonaria Goffi


    Full Text Available Primary infection of immunocompetent individuals does not lead to complication, but CMV is a major clinical problem in transplant recipients.Thus, it is important to use sensitive and specific diagnostic techniques to rapidly and accurately detect CMV infection and identify patients at risk of developing CMV disease. In the present study, CMV infection after bone marrow transplantation was monitored by two molecular biology assays: COBAS AMPLICOR CMV MONITOR and NUCLISENS CMV m-RNA pp67. CMV m-RNApp67 assay was found to show positivity later than CMV-DNA MONITOR. The increment of number of copies/ml agreed with the clinical symptoms of CMV infection. The quantitative results of the CMV MONTOR assay was more helpfull to select an antiviral strategy than the qualitative results of NUCLISENS m-RNA pp67 assay.

  17. Alteration in sample preparation to increase the yield of multiplex Polymerase Chain Reaction assay for diagnosis of genital ulcer disease

    G Rao


    Full Text Available Purpose: Genital Ulcer Disease (GUD is common sexually transmitted infection (STI. Multiple studies have shown that GUDs are strongly associated with the transmission and the acquisition of HIV infection. An accurate diagnosis of common etiology of GUD namely Herpes, syphilis and Chancroid is possible using Multiplex PCR (M-PCR. However, frequent presence of Polymerase Chain Reaction inhibitors in the ulcer swab specimen limits the performance of the assay. In order to overcome this problem, alternative specimen preparation method was used. Materials and Methods: To determine the common etiology, GUD specimens obtained under an STI operations research study were tested with M-PCR after the samples were prepared using Roche Amplicor specimen preparation kit. PCR inhibiting samples were identified from that, which showed negative results. These samples were subjected to phenol-chloroform extraction and ethanol precipitation before the conduct of M-PCR on them. Results: Of the 237 GUD specimens tested, in 145 etiologies could be detected, whereas 92 samples were found negative. Further spiking with one of the target DNA, 128 of the negative samples were found to contain the inhibitors. These 126 samples were then subjected to phenol chloroform extraction and ethanol precipitation followed by M-PCR. Using this method for sample preparation, etiology could be determined in 46 (23% additional samples. This success rate of altered sample preparation method has been lower than that has reported. Conclusion: The results indicate that sample preparation using phenol chloroform extraction and ethanol precipitation, prior to M-PCR helps to eliminate the inhibitors and increase the yield of the assay. However, being a laborious procedure, it may be used for samples giving negative results after the screening by Roche Amplicor specimen preparation kit.

  18. Effect of Antiviral Therapy on Serum Activity of Angiotensin Converting Enzyme in Patients with Chronic Hepatitis C

    Husic-Selimovic, Azra; Sofic, Amela; Huskic, Jasminko; Bulja, Deniz


    Introduction: Renin-angiotenzin system (RAS) is frequently activated in patients with chronic liver disease. Angiotenzin - II (AT-II), produced by angiotenzin converting enzyme (ACE), has many physiological effects, including an important role in liver fibrogenesis. Combined antiviral therapy with PEG-IFN and ribavirin besides its antiviral effect also leads to a reduction in liver parenchyma fibrosis. Aim of the study: Determining the value of ACE in serum of patients with chronic hepatitis C before and after combined antiviral therapy, as well as the value of ACE activities in sera of the control group. Materials and methods: We studied 50 patients treated at Gastroenterohepatology Department, in the time-period of four years. Value of ACE in serum was determined by Olympus AU 400 device, with application of kit “Infinity TN ACE Liquid Stable Reagent”. HCV RNA levels in sera were measured by real time PCR. HCV RNA test was performed with modular analysis of AMPLICOR and COBAS AMPLICOR HCV MONITOR test v2.0, which has proved infection and was used for quantification of the viruses and monitoring of the patients’ response to therapy. Liver histology was evaluated in accordance with the level of necroinflammation activity and stage of fibrosis. Results: Serum activities of ACE in chronic hepatitis C patients is statistically higher than the values in the control group (p=0.02). Antiviral therapy in chronic hepatitis C patients statistically decreases serum activities of ACE (p= 0.02) and indirectly affects fibrogenesis of the liver parenchyma. Correlation between ACE and ALT activity after the therapy was proved (0.3934). Conclusion: Our findings suggest that the activity of ACE in serum is a good indirect parameter of the liver damage, and could be used as an indirect prognostic factor of the level of liver parenchyma damage. Serum activity of ACE can be used as a parameter for non-invasive assessment of intensity of liver damage. PMID:27147779

  19. Viral blips during long-term treatment with standard or double dose lamivudine in HBe antigen negative chronic hepatitis B


    AIM: To evaluate safety and effect on hepatitis B virus (HBV) suppression of a long-term treatment with lamivudine (LAM) at standard (100 mg/d) or double (200 mg/d) dose in chronic hepatitis B. METHODS: This was a case study with matched controls (1:3) in patients with chronic hepatitis B with anti-Hbe antibodies. RESULTS: Twelve patients received LAM 200 mg/d and 35 LAM 100 mg/d, for a median of 28 mo. A primary response (PR; I.e. Negative HBV-DNA with Amplicor assay) was achieved in 100% of LAM-200 patients and 83% of LAM-100 patients. A virological breakthrough occurred in 16.7 and 24.7%, respectively, of the PR-patients, with the appearance of typical LAM resistance mutations in all but one patient. Viremia blips (I.e. Transient HBV-DNA below 80 IU/Ml in patients who tested negative at Amplicor assay) were detected using a real time polymerase chain reaction (PCR) and occurred in seven out of nine patients with subsequent BT and in four out of 32 patients with end-of-study response (77.7% vs 12.5%; P = 0.001) at chi-square test). At the end of the study, 51.4% of LAM-100 patients and 83.3% of LAM-200 patients had remained stably HBV-DNA negative. Double-dose LAM was well tolerated. CONCLUSION: Long-term treatment of anti-Hbe positive chronic hepatitis B with double dose lamivudine causes a more profound and stable viral suppression as compared to conventional treatment.

  20. Hepatitis C virus RNA assays: a comparison of SuperQuant and Monitor.

    Hadziyannis, E; Hadziyannis, A; Yen-Lieberman, B; Kiwi, M L; Hodnick, S; Spanou, F; Starkey, C; Younossi, Z M


    Hepatitis C RNA testing has been used extensively to assess the efficacy of antiviral therapy and has increasingly become an integral part of clinical management of patients with chronic hepatitis C. A variety of commercially available hepatitis C virus (HCV) RNA tests are used to detect HCV RNA qualitatively or quantitatively. These commercial tests have fundamental differences that are reflected on the values they generate. We compared two widely used assays, HCV SuperQuant (SQ) and Amplicor HCV Monitor (M1 and M2), in sera of patients with chronic hepatitis C. A total of 506 sera from 79 patients were tested with both assays. The data were logarithmically transformed and analyzed by linear regression and measurement of agreement. Two hundred thirty-eight sera had HCV RNA values within the dynamic range of both assays. The correlation between the assays was fair, with a correlation coefficient (r) of 0.699. Overall, SQ generated higher values than M1 with a mean difference of 0.558 log (SD = 0.624). One hundred ninety-four (38%) and 121 (24%) of the sera were below the dynamic range of M1 and SQ, respectively. Seventy-three sera, undetectable by M1, were positive by SQ. The Amplicor HCV Monitor 2.0 (M2) was performed in 66 sera. All were positive by SQ and M2, but only 38 were within the dynamic range of M1. The correlations between these tests were good (r = 0.68-0.78), but the agreement was rather poor. In conclusion, this study confirms that both SQ and M2 are more sensitive than M1. Additionally, our results show rather poor agreements between these assays. The recent attempts in standardizing the reporting of these assays should make their results more easily interchangeable. PMID:11418790

  1. MSD grant läks Narva / MSD Teaduskeskus


    MSD (Merck Sharpe & Dohme) regionaalsete grantide komitee kuulutas välja konkursi võitja, selleks osutus Narva MTÜ Rehabilitatsioonikeskus "Sind ei jäeta üksi". 2,4 mln kroonine grant on mõeldud noortele HIV-i ja narkootikumide alaseks ennetustööks Ida-Virumaal


    G. A. Ryasanova; I. M. Fazulzyanova


    Abstract. In HIV-infected patients, the process of antibody production to certain HIV-I structural proteins proceeds in differential manner, depending on the antigen localization. Upon progression of the disease, an increased ratio of antibodies to env surface glycoproteins is found, along with decreased percentage of antibodies to gag gene proteins.

  3. Üle poole noortest istub vabal ajal baaris / Merike Enniko-Väljaots

    Enniko-Väljaots, Merike


    Lääne-Virumaa noorte hulgas korraldatud uurimus tõi välja teismeliste sõltuvusaine lembuse ja teadmatuse HIV-i ja AIDS-i osas. Lisaks diagramm: Vanus sõltuvusainete esmasel proovimisel sugude lõikes (%)

  4. Maret Maripuu : HIV-haigete raviks jätkub raha ka järgmisel aastal / Maret Maripuu ; interv. Ille Grün-Ots

    Maripuu, Maret, 1974-


    Sotsiaalministri Maret Maripuu sõnul leiab HIV-i vältimise vajalikkus üha rohkem kõlapinda kogu ühiskonnas ja poolele teele seisma jääda ei tohi. Kommenteerib Lääne-Tallinna Keskhaigla nakkuskeskuse juhataja Kai Zilmer

  5. Expanded breadth of the T-cell response to mosaic HIV-1 envelope DNA vaccination

    Korber, Bette [Los Alamos National Laboratory; Fischer, William [Los Alamos National Laboratory; Wallstrom, Timothy [Los Alamos National Laboratory


    An effective AIDS vaccine must control highly diverse circulating strains of HIV-1. Among HIV -I gene products, the envelope (Env) protein contains variable as well as conserved regions. In this report, an informatic approach to the design of T-cell vaccines directed to HIV -I Env M group global sequences was tested. Synthetic Env antigens were designed to express mosaics that maximize the inclusion of common potential Tcell epitope (PTE) 9-mers and minimize the inclusion of rare epitopes likely to elicit strain-specific responses. DNA vaccines were evaluated using intracellular cytokine staining (ICS) in inbred mice with a standardized panel of highly conserved 15-mer PTE peptides. I, 2 and 3 mosaic sets were developed that increased theoretical epitope coverage. The breadth and magnitude ofT-cell immunity stimulated by these vaccines were compared to natural strain Env's; additional comparisons were performed on mutant Env's, including gpl60 or gpl45 with or without V regions and gp41 deletions. Among them, the 2 or 3 mosaic Env sets elicited the optimal CD4 and CD8 responses. These responses were most evident in CD8 T cells; the 3 mosaic set elicited responses to an average of 8 peptide pools compared to 2 pools for a set of3 natural Env's. Synthetic mosaic HIV -I antigens can therefore induce T-cell responses with expanded breadth and may facilitate the development of effective T -cell-based HIV -1 vaccines.

  6. Nucleic acid amplification technology screening for hepatitis C virus and human immunodeficiency virus for blood donations

    To investigate the performance of the commercial Roche COBAS AmpliScreen assay, and demonstrate whether the COBAS AmpliScreen human immunodeficiency virus-1 (HIV-1) test, v1.5, and COBAS AmpliScreen hepatitis C virus (HCV) v 2.0 for screening for HIV-1 and HCV RNA in the donated blood units from which plasma mini pools were collected, by nucleic acid amplification technology (NAT), could detect the positive pools and reduce the risk of transmission of infections for those routinely tested by serological assays. The study was performed on 3288 plasma samples collected from blood donors in a period of 13 months, from August 2004 to August 2005, at Al-Hada Armed Forces Hospital, Molecular Pathology Laboratory, Taif, Kingdom of Saudi Arabia. The samples were tested by the reverse transcriptase polymerase chain reaction (RT-PCR) after RNA extraction (this represents the major method in NAT assays), in parallel with the routine serological testing to detect qualitatively for HIV-1 and HCV. The NAT assays that include an automated COBAS AmpliPrep system for RNA extraction and COBAS Amplicor Analyzer using AmpliScreen kits for RT-PCR assays, and the routine serological screening assays for the detection of the HIV-1 and HCV RNA in the plasma samples from the blood donors have shown to be a reliable combination that would meet our requirements. The collected data further confirms the results from the serological assays and enables us to decrease the residual risk of transmission to a minimum with the finding of no seronegative window period donation. The results demonstrate that out of 3288 samples, the percentages of RT-PCR (NAT) negative blood donations that were also confirmed as seronegative were 99% for HCV, and 99.1% for HIV-1. The modified combined systems (automated COBAS AmpliPrep system for RNA extraction and COBAS Amplicor Analyzer using AmpliScreen kits for RT-PCR assays) for NAT screening assays has allowed the release of all blood donations supplied in the

  7. Does chronic alcohol use by HIV-infected patients on d4T/3TC/NVP drug regimen effect the HIV viral load and what is the therapeutic window of the drugs, CD4+ count and WBC count in patients with high viral load during the 9 months period of follow up?

    Godfrey S. Bbosa


    Full Text Available The study investigated the effects of chronic alcohol use on HIV viral load in HIV-infected patients on d4T/3TC/NVP drug regimen during 9 months follow up period. It also determined plasma drug concentrations of d4T, 3TC and NVP; CD4+ and WBC counts for patients with high HIV viral load. A case-control study using repeated measures with serial measurements was used. A total of 41 patients (20 alcohol group and 21 control group were screened for alcohol use using WHO AUDIT tool and chronic alcohol use biomarkers. Blood sampling was done at 3 month intervals for a period of 9 months. HIV viral load was determined using Roche Amplicor HIV-1 monitor test, version 1.5 (Amplicor. The d4T, 3TC and NVP concentrations were determined by Shimadzu Class-VPTM HPLC Chromatography data system version 6.1. The CD4+ cell count was determined using FACSCalibur flow cytometer. The WBC was determined using automated hematological Coulter CBC-5 Hematology Analyzer system. Results show that % patients with HIV viral load ≥400 copies/ml in control group was highest (23.8%, n=5 at 3 month while in chronic alcohol use group, it was at 0 month (35%, n=7 for both WHO AUDIT tool and chronic alcohol-use biomarkers groups. Generally patients with high viral load ≥400 copies/ml was observed in chronic alcohol use as compared to control group in both WHO AUDIT tool and biomarkers group despite of patients having high steady state d4T, 3TC and NVP plasma drug concentrations in circulation that is available to suppress HIV virus. The high viral load could be associated with the emergence of resistance of the HIV virus and these patients generally had a low CD4+ cell count. Some of these patients had no detectable d4T plasma drug concentrations in circulation and most of them with high viral load had sub-therapeutic NVP plasma drug concentrations in their blood circulation. Chronic ethanol use by HIV-infected patients on d4T/3TC/NVP drug regimen increased HIV viral load and

  8. Prevalence of Hepatitis C Virus in the Population of Albania for the Period 2007-2010

    Hysaj Vila Brunilda


    Full Text Available BACKGROUND: Hepatitis C is a blood-borne, infectious, viral disease that is caused by a hepatotropic virus called Hepatitis C virus (HCV. AIM: The aim of this study is to determine the prevalence of active HCV infection (HCV–RNA in the cases that were anti-HCV positive. MATERIAL AND METHODS: Plasma of 301 high-risk for HCV infection consecutive from University Hospital Centre “Mother Theresa” Tirana-Albania, during January 2007 to December 2010 was included in this study. To identify the presence of HCV RNA, the samples were examined by Cobas Amplicor HCV test (qualitative method. RESULTS: From 301 samples analyzed in total, 214 of them resulted positive for the presence of HCV-RNA's, corresponding to a prevalence of 71.1%, with 95% CI interval [65.8 - 75.9] for value of χ2 = 52.7 p value 25 years with a significant difference with other age groups for p value 25 years.

  9. Factors influencing a low rate of hepatitis C viral RNA clearance in heroin users from Southern China

    ShengHan Lai; Jin-Bing Zhang; Wei Liu; Jie Chen; Xiao-Fang Yu


    AIM: To study the virological and host factors influencing hepatitis C infection outcomes in heroin users in southern China.METHODS: HCV RNA and associated factors were analyzed among 347 heroin users from Guangxi Zhuang Autonomous Region, southern China who were hepatitis C virus (HCV) EIA positive for two or more consecutive visits.RESULTS: Using the COBAS AMPLICOR HCV TEST, a remarkably low HCV RNA negative rate of 8.6% was detected. After multivariate logistic regression analysis, HCV RNA clearance was significantly associated with the presence of HBsAg (OR = 8.436, P < 0.0001), the lack of HIV-1 infection (OR = 0.256, P = 0.038) and age younger than 25 (OR = 0.400, P = 0.029).CONCLUSION: Our study suggests HCV infection among Chinese heroin users results in high levels of viral persistence even amidst factors previously found to enhance viral clearance. Prospective studies of a possible genetic component within the Chinese population and the pathogenicity of non-genotype 1 HCV infections are needed.

  10. Lyophilized standards for the calibration of real time PCR assay for hepatitis C virus RNA

    WANG Lu-nan; WU Jian-min; DENG Wei; SHEN Zi-yu; CHEN Wen-xiang; LI Jin-ming


    Background Since October 1997, an international standard for hepatitis C virus (HCV) nucleic acid amplification technology assay, 96/790, has been available. We compared a series of lyophilized standards with known HCV RNA concentrations against the international standard in fluorescence quantitative PCR detection.Methods A series of lyophilized sera were calibrated by ROCHE COBAS AMPLICOR HCV Monitor test against the international standard and sent to various manufacturers to analyse the samples using their own kits.Then calibration curves from the series were compared with that obtained from the external standard calibration curve with the manufacture's series.Results The standard calibration curve with the series of lyophilized serum showed an excellent correlation(R2>0.98), slope and intercept that were similar to those from the manufacture's series. When the standard calibration curve from the series of lyophilized standards were used to define the values of the given sample,lower coefficients of variation between kits from different manufactures were obtained.Conclusion The results showed that the lyophilized standards could be used to setup the standard calibration curve for clinical HCV RNA quantitative PCR detection.

  11. Prevalence of Chlamydia trachomatis in women attending sexually transmitted disease clinics in the Colombo district, Sri Lanka

    Gunasekera Henadira Appuhamilage Kamani Mangalika


    Full Text Available Background: In Sri Lanka little is known about the prevalence of Chlamydia trachomatis (CT infection. Objective was to determine the prevalence of CT in female patients attending sexually transmitted disease (STD clinics in the Colombo district. Materials and Methods: A descriptive cross-sectional study was carried out for the prevalence of CT in all female patients (n = 168 more than 18 years of age, attending two STD clinics in the Colombo district from January to May 2012. Endocervical swabs were collected and tested for CT using the Amplicor CT/NG polymerase chain reaction assay. Results: Prevalence of CT in females attending the STD clinics in the Colombo district was 8.3%. Mean age of those infected with CT was 32.9 years (SD ± 8.2. Majority of females with CT infections were Sinhalese and married. There was no significant association with age, ethnicity or being married or not. Females who did not attend school, or had their education only up to Grade 5 were significantly found to have six times the risk of having CT infection (95% CI = 1.8-22.6. A significant association was found with number of sexual partners but not with commercial sex work or past history of STD. Conclusions: Prevalence of CT was moderately high in this population.

  12. Persistence of hepatitis C virus in a white population: associations with human leukocyte antigen class 1.

    Fanning, Liam J


    The aim of this study was to define novel associations between human leukocyte antigen (HLA) class 1 alleles and persistence or clearance of the hepatitis C virus (HCV) in a white population. All individuals in the study were seropositive for anti-HCV antibodies. Viral status was determined by the Roche HCV Amplicor test. HLA-A, -B, -C allelic group profile was molecularly defined by reverse line probe hybridization. The strongest individual allelic group associations with persistent HCV infection were HLA A*11 (p = 0.044) and Cw*04 (p = 0.006). However, only the HLA C*04 association survived correction for multiple comparisons. Further analysis of alleles in linkage with HLA Cw*04 revealed that the haplotype HLA A*11, Cw*04 was present in 11 individuals, 10 of whom were viremic (p = 0.05). No gene dosage effect was observed. No association between HLA class 1 allelic groups and aviremia and virus load was evident in this white population. HLA B*44 is associated with low virus load in human immunodeficiency virus disease, but this association was not evident in this HCV-infected population. Novel HLA class 1 alleles associated with persistence of HCV have been identified.

  13. Strategy for the maximization of clinically relevant information from hepatitis C virus, RT-PCR quantification.

    Levis, J


    BACKGROUND: The increasing clinical application of viral load assays for monitoring viral infections has been an incentive for the development of standardized tests for the hepatitis C virus. OBJECTIVE: To develop a simple model for the prediction of baseline viral load in individuals infected with the hepatitis C virus. METHODOLOGY: Viral load quantification of each patient\\'s first sample was assessed by RT-PCR-ELISA using the Roche MONITOR assay in triplicate. Genotype of the infecting virus was identified by reverse line probe hybridization, using amplicons resulting from the qualitative HCV Roche AMPLICOR assay. RESULTS: Retrospective evaluation of first quantitative values suggested that 82.4% (n=168\\/204) of individuals had a viral load between 4.3 and 6.7 log(10) viral copies per ml. A few patients (3.4%; n=7\\/204) have a serum viremia less than the lower limit of the linear range of the RT-PCR assay. Subsequent, prospective evaluation of hepatitis C viral load of all new patients using a model based on the dynamic range of viral load in the retrospective group correctly predicted the dynamic range in 75.9% (n=33\\/54). CONCLUSION: The dynamic range of hepatitis C viremia extends beyond the linear range of the Roche MONITOR assay. Accurate determination of serum viremia is substantially improved by dilution of specimens prior to quantification.

  14. Stable hepatitis C virus RNA detection by RT-PCR during four days storage

    Horsmans Yves


    Full Text Available Abstract Background Suboptimal specimen processing and storage conditions of samples which contain hepatitis C virus (HCV RNA may result in a decline of HCV RNA concentration or false-negative results in the detection of HCV RNA in serum. We evaluated the stability of HCV RNA in serum and clotted blood samples stored at room temperature or at 4°C for 4 days with the aim of optimizing the standard procedures of processing and storage of samples. Methods Blood from five HCV RNA positive patients was collected in tubes with and without separator gel, centrifuged 1 or 6 hours after collection. Samples were then left 6, 24, 48, 72 or 96 h at room temperature (21.5 – 25.4°C or at 4°C before determining their HCV RNA level using the COBAS AMPLICOR HCV MONITOR Test, vs 2.0 (Roche Diagnostic Systems. Results The logarithm of the HCV RNA level measurements remained within a 0.3 value of the means for 4 days at both temperatures (room temperature or 4°C. Conclusions We conclude that blood samples may be collected and aliquoted within 6 h of collection and can be stored at 4°C for 72 hours as proposed by the manufacturer without significant differences in measured HCV RNA level. Our results indicate that lapses in this scheme may still yield reliable results.

  15. Genotyping Pattern Among Iranian HCV Positive Patients

    A Sarafnejad


    Full Text Available Background: Successful treatment to eliminate HCV RNA depends on the identified genotype. In the present study, we compared the frequency of different HCV genotypes, during four years study (2004 till 2008.Methods: Sera specimens were received from 16 provinces of Iran. We used High Pure Viral Nucleic Acid Purification kit for extraction and samples were tested with improved form of RT-PCR technique. HCV genotypes were determined using Amplisense PCR kit and Amplicor HCV Monitoring Version 2 test utilized a reverse transcription (RT-PCR approach to quantitative HCV RNA. Two hundreds six HCV positive specimens were entered to the study out of 389 tested samples.Results: Type 3a was the most frequent type (46.6%, followed by type 1 (including 1a and 1b with 25.73% and 17.47% for each respectively with 43.2%. Looking through collected results of the four years study confirmed the rate of HCV infection in those single genotypes 1b, 3a were slightly increased from 12.22% and 38.88% in the first year to 18.66 and 46.51% in the fourth year of the study period.Conclusion: The analyzed data proved that some patients were infected with two different types. High viral load was also more correlated to genotype 1 than other types.

  16. Evaluation of the screening test results before marriage

    Süleyman Durmaz


    Full Text Available Objectives: Human immunodeficiency virus (HIV, Hepatitis B and Hepatitis C viruses and Treponema pallidum are parenterally and sexually transmitted infection agents. Screening test is made before marriage to pre-marital couples legally under the relevant legislation and legal procedures in our country; applicants are evaluated in terms of sexually transmitted diseases. The aim of this study is to evaluate pre-marital test results for HBsAg, anti-HCV, anti-HIV I/II and Treponema pallidum.Materials and methods: To make screening test before marriage, randomized 117 patients who were applied to Kızıltepe General Hospital of Infectious Diseases and Clinical Microbiology, were included in this study between January 2011 and March 2011. Of these patients, 64 were women (average age 24.7±5.7, and 55 were males (mean age 24.7±4.7. HBsAg, anti-HCV and anti-HIV I/II tests of the patients were studied by macro-ELISA device (ECIQ Vitros, Ortho Clinical Diagnostics, USA, screening of anti-Treponema pallidum IgG, IgA and IgM antibodies were studied by immunochromatographic rapid test (syphilis syphilis 3.0, Standard Diagnostics, inc. Korea.Results: Of the 119 patients, five patients (4.2% were positive for HBsAg (3 male and 2 female. Anti-HCV, anti-HIV I/II and anti-Treponema pallidum antibodies were negative in all patients.Conclusion: HBsAg test result which was obtained in present study has been found consistent with HBsAg positivity rate in our region. As a result of screening test that was done before marriage will continue to believe that the increased importance of the prevention of sexually transmitted diseases. J Clin Exp Invest 2011; 2 (3: 292-294.

  17. Hvordan kan sykepleier bidra til at HIV-positive mestrer sykdommens utfordringer i hverdagen?

    Wilhelmsen, Tonje Johansen


    Bakgrunn: HIV er en sykdom som er forbundet med skam, skyldfølelse og frykt. På verdensbasis beregnes det at 34 millioner mennesker er smittet av HIV. I Norge smittes mellom 200-250 personer årlig av HIV, og i 2010 var det 10 mennesker som omkom som følge av sykdommen. I samfunnet vårt er HIV en tabubelagt og stigmatisert sykdom. Mange som lever med sykdommen lever dermed et «hemmelig liv». Problemstilling: «Hvordan kan sykepleier bidra til at HIV-positive mestrer sykdommens utfordringer i...

  18. The initial antibody response to HIV-1: induction of ineffective early B cell responses against GP41 by the transmitted/founder virus

    Chavez, Leslie L [Los Alamos National Laboratory; Perelson, Alan [Los Alamos National Laboratory


    A window of opportunity for immune responses to extinguish HIV -1 exists from the moment of transmission through establishment of the latent pool of HIV -I-infected cells. A critical time to study the initial immune responses to the transmitted/founder virus is the eclipse phase of HIV-1 infection (time from transmission to the first appearance of plasma virus) but, to date, this period has been logistically difficult to analyze. Studies in non-human primates challenged with chimeric simianhuman immunodeficiency virus have shown that neutralizing antibodies, when present at the time of infection, can prevent virus infection.

  19. StochKit-FF: Efficient Systems Biology on Multicore Architectures

    Aldinucci, Marco; Bracciali, Andrea; Liò, Pietro; Sorathiya, Anil; Torquati, Massimo


    The stochastic modelling of biological systems is an informative, and in some cases, very adequate technique, which may however result in being more expensive than other modelling approaches, such as differential equations. We present StochKit-FF, a parallel version of StochKit, a reference toolkit for stochastic simulations. StochKit-FF is based on the FastFlow programming toolkit for multicores and exploits the novel concept of selective memory. We experiment StochKit-FF on a model of HIV i...

  20. Chlamydia trachomatis infection and sexual behaviour among female students attending higher education in the Republic of Ireland.

    O'Connell, Emer


    BACKGROUND: There are no prevalence data on Chlamydia trachomatis relating to female students attending higher education available for the Republic of Ireland. This information is required to guide on the necessity for Chlamydia screening programmes in higher education settings. This research aimed to determine the prevalence of and predictive risk factors for Chlamydia trachomatis genital infection among female higher education students in Ireland. METHODS: All females presenting during one-day periods at Student Health Units in three higher education institutions in two cities in the Republic of Ireland were invited to participate. Participants completed a questionnaire on lifestyle and socio-demographic factors and provided a urine sample. Samples were tested for C. trachomatis DNA by a PCR based technique (Cobas Amplicor, Roche). To examine possible associations between a positive test and demographic and lifestyle risk factors, a univariate analysis was performed. All associations with a p value < 0.05 were included in a multivariate logistic regression analysis. RESULTS: Of the 460 sexually active participants 22 tested positive (prevalence 4.8%; 95% CI 3.0 to 7.1%). Variables associated with significantly increased risk were current suggestive symptoms, two or more one-night stands and three or more lifetime sexual partners. The students displayed high-risk sexual behaviour. CONCLUSION: The prevalence of C. trachomatis infection and the lack of awareness of the significance of suggestive symptoms among sexually experienced female students demonstrate the need for a programme to test asymptomatic or non-presenting higher education students. The risk factors identified by multivariate analysis may be useful in identifying those who are most likely to benefit from screening. Alcohol abuse, condom use, sexual behaviour (at home and abroad) and, knowledge of sexually transmitted infections (STIs) (including asymptomatic nature or relevant symptoms) were

  1. Indeterminate RIBA results were associated with the absence of hepatitis C virus RNA (HCV-RNA in blood donors

    Felicidade Mota Pereira


    Full Text Available Introduction: Hepatitis C virus (HCV infection is diagnosed by the presence of antibodies and is supplemented by confirmatory testing methods, such as recombinant immunoblot assay (RIBA and HCV-RNA detection. This study aimed to evaluate the efficacy of RIBA testing to diagnose HCV infection in blood donors positive for anti-HCV antibodies. Methods: A total of 102 subjects positive for anti-HCV determined by enzyme-linked immunosorbent assay (ELISA at the Hematology and Hemotherapy Foundation of Bahia (HEMOBA were later assessed with new samples using the Abbott Architect anti-HCV test (Abbott Diagnostics, Wiesbaden, Germany, the RIBA III test (Chiron RIBA HCV 3.0 SIA, Chiron Corp., Emeryville, CA, USA, the polymerase chain reaction (PCR; COBAS® AMPLICOR HCV Roche Diagnostics Corp., Indianapolis, IN, USA and line probe assay (LiPA - Siemens, Tarrytown, NY, USA genotyping for HCV diagnosis. Results: Of these new samples, 38.2% (39/102 were positive, 57.8% (59/102 were negative and 3.9% (4/102 were indeterminate for anti-HCV; HCV-RNA was detected in 22.5% (23/102 of the samples. RIBA results were positive in 58.1% (25/43, negative in 9.3% (4/43 and indeterminate in 32.6% (14/43 of the samples. The prevailing genotypes were 1 (78.3%, 18/23, 3 (17.4%, 4/23 and 2 (4.3%, 1/23. All 14 samples with indeterminate RIBA results had undetectable viral loads (detection limit ≤50 IU/mL. Of these samples, 71.4% (10/14 were reevaluated six months later. Eighty percent (8/10 of these samples remained indeterminate by RIBA, and 20% (2/10 were negative. Conclusions: In this study, individuals with indeterminate RIBA results had no detectable HCV-RNA.

  2. HPV: screening and prevalence of genotypes in the lower-Molise

    Mariangela Spinosa


    Full Text Available HPV is a double-stranded DNA virus. It is a sexually transmitted virus about 75% of women contact it throughout their lives.Among the 100 classified genotypes about 40 can infect the genital mucosa. Depending on the type of lesion can be identified genotypes with a “low risk” and “high risk”.These are associated with cervical dysplasia and carcinoma of the uterine cervix. Il “Pap smear” is the cytological test that highlights the changes of the cervical cells. L’’HPV-DNA-PCR can detect DNA and determine the infecting viral genotype. It is a susceptible, specific and not invasive. test.Aim of our work was to evaluate a screening program aimed at testing the prevalence of HPV genotypes in the lower-Molise. We have examined 339 samples obtained from cervical swabs of women aged 18-45 years.Was used-Amplicor HPV test (Roche a qualitative method for detection of 13 high-risk genotypes. Genotype was identified using the Linear Array HPV-Genotyping (Roche method. Among 339 women tested, 292 (86% were negative, 47 (14% positive. It has been performed genotyping on 47 positiv samples. 16 and 18 were the prevalent genotypes (average 8%.There was a lower prevalence between 3% and 2% among the other genotypes. These results allow us to draw some considerations while taking into account the limited number of samples.The frequency of positive HPVDNA test it is very high.The HPV-DNA testing is a valuable aid in diagnosis by HPV alongside the Pap Test. The prevalence of genotypes found in the Low-Molise is consistent with data reported in literature.The genotypes 16 and 18 have a higher frequency, taking into account that these genotypes are responsible for 70% of cervical cancer, the determination may be a useful aid in the diagnosis and prevention.

  3. Síndrome da meningite asséptica por enterovírus e Leptospira sp em crianças de Salvador, Bahia

    Silva Hagamenon R.


    Full Text Available Objetivando verificar a freqüência de enterovírus (EV, leptospiras e arbovírus como agentes causais da síndrome da meningite asséptica (SMA, em períodos não-epidêmicos, e comparar os pacientes com e sem diagnóstico etiológico determinado, foram selecionados 112 pacientes de idade entre 3 meses e 15 anos, com suspeita clínica de SMA, referenciados para Hospital Couto Maia, especializado em Doenças Infecciosas e Parasitárias (Salvador, Bahia, Em 44,6% (n=50 a etiologia foi determinada: enterovírus em 37,7% (n=42 dos casos, pelo teste de PCR Amplicor, por cultura do líquor e/ou de fezes; a Leptospira sp. em 7,1% (n=8, pelo método da micro-aglutinação, e nenhum caso de arbovírus foi detectado (inibição da hemaglutinação passiva. Entre os 14 enterovírus dos 22 isolados, foram identificados seis diferentes sorotipos, sendo o Echovirus-4 predominante (27,2%; 6/22 entre outros (Coxsackie B2, B3, B6 e B9; EV 71. Conclui-se que, os enterovírus foram os agentes mais freqüentes, e que a leptospirose deve ser lembrada no diagnóstico diferencial da SMA. Uma vez que as características clínicas e liquóricas dos pacientes dos grupos com e sem determinação do agente etiológico foram semelhantes, pode-se supor que o diagnóstico presuntivo de SMA é de provável etiologia viral ou pela leptospira.

  4. “That’s True Love:” Lived Experiences of Puerto Rican Perinatally HIV-Infected Youth within Their Families’ Context

    Georgina Silva-Suárez


    Full Text Available The burden of HIV affects not only HIV-infected patients but also their families and caregivers. It is also known that family support is crucial for people living with HIV. A qualitative study was conducted to explore the life experiences, within the family context, of perinatally HIV-infected (pHIV-I youth in Puerto Rico. Twenty in-depth interviews were performed and audio-recorded. Within the family context, study participants experienced acceptance, love and support but also stigma and discrimination. They reported that family is an essential component in their lives and treatment. Losing one or both parents at a young age was considered more difficult than having HIV. Most participants who lost their parents lived with other family members. This was a challenging situation for both pHIV-I youth and their caregivers. Participants described their healthcare providers as part of their families and would like to keep in touch as they transition to adult care. Despite the challenges, participants expressed a desire to have children. Services targeted to this population should stress social support, incorporate family members into the medical process, provide special guidance and support while transitioning to adult care, and provide them with the latest information regarding HIV and reproductive options.

  5. Photosensitized inactivation of infectious blood-borne human parasites

    Judy, Millard M.; Sogandares-Bernal, Franklin M.; Matthews, James Lester


    Blood-borne viruses and protozoan parasites that are infectious to humans pose risk world-wide of infection transmission through blood and blood product transfusion. Blood-borne infectious viruses include human immunodeficiency virus (HIV-I), which causes AIDS; hepatitis C virus, which can cause chronic hepatitis; and cytomegalovirus, which can be dangerous to immunocompromised patients, e.g., the newborn, transplant recipients, and AIDS patients. Infectious blood-borne protozoan parasites include Trypanosoma cruzi, which causes Chagas' disease, endemic throughout Central and South America; the Trypanosoma species causing African sleeping sickness endemic in Central Africa; and Plasmodium falciparum, which causes malignant and increasingly drug- resistant human malaria prevalent throughout the tropics. Some researchers have focused on using photosensitizers to inactivate HIV-I and other viruses in whole blood, packed red cells, and platelet concentrates without compromising blood product function. Our group previously has reported photosensitized in vitro inactivation of P. falciparum and the mouse malaria organism Plasmodium berghei in whole blood using hematoporphyrin derivative (HPD) and of T. cruzi using benzoporphyrin derivatives BPDMA and BPDDA, dihematoporphyrin ether (DHE), and hydroxyethylvinyldeuteroporphyrin (HEVD). These results suggest that continued investigation is warranted to evaluate the potential for photosensitized inactivation of blood-borne parasites in blood banking.

  6. [Role of line immunoassay in the diagnosis of early HIV infection: a diagnostic case].

    Soylar, Muhammed; Altuğlu, Imre; Sertöz, Rüçhan; Gökengin, Deniz


    Combined p24 antigen-HIV antibody fourth-generation assays that identify most of the early HIV infections have been used extensively worldwide for several years. This poses challenges for the traditional algorithm of line immunoassay (LIA) confirmation. LIA tests are useful methods with their high specificity and their ability to differentiate HIV-1 from HIV-2, but they are reactive days after the fourth generation enzyme immunoassays. With acute HIV infection, high levels of infectious virus are detectable in serum and genital secretions. The rate of transmission during acute HIV infection is higher than the established HIV infection, for this reason, new HIV testing strategies need to focus on sensitivity, especially for this highly contagious phase immediately after infection. Serum sample of a patient sent to Ege University Hospital Clinical Virology Laboratory was repeatedly reactive with low signal/cutoff ratios with two different commercial fourth generation enzyme immunoassays (Architect HIV Ag/Ab Combo Reagent Kit, Abbott, Germany and Vidas HIV Duo Quick, Biomerieux, France). The sample was non-reactive with the LIA (INNO-LIA HIV I/II Score, Innogenetics, Belgium) and HIV RNA (RealTime HIV-I Amplification Reagent Kit, Abbott, USA) result was positive (4.1 x 10(5) copies/ml). With the presentation of this case, the role of LIA in the diagnosis of early HIV infection and its place in test algorithms were questioned. PMID:23971936

  7. 033 免疫色谱法快速检测HIV抗体的评价


    @@由于HIV感染的发生与传播呈上升趋势,急需建立一种快速、简便、成本低、灵敏度高、特异性好的抗HIV抗体检测方法.本文对Determine HIV-1/HIV-2法(即免疫色谱法)与明胶颗粒凝集法(PA)、乳胶凝集试验、免疫色谱纸电法(HemaStrip HIV-1/HIV-2)进行了对比,证实Determine HIV-i/HIV-2具备以上优点.

  8. NKT cells in HIV-1 infection


    Natural killer T (NKT) cells are a unique T cell population that have important immunoregulatory functions and have been shown to be involved in host immunity against a range of microorganisms. It also emerges that they might play a role in HIV-1 infection, and therefore be selectively depleted during the early stages of infection. Recent studies are reviewed regarding the dynamics of NKT depletion during HIV-I infection and their recovery under highly active antiretrovirai treatment (HAART). Possible mechanisms for these changes are proposed based on the recent developments in HIV pathogenesis. Further discussions are focused on HIV's disruption of NKT activation by downregulating CDId expression on antigen presentation cells (APC). HIV-1 protein Nefis found to play the major role by interrupting the intraceilular trafficking of nascent and recycling CDId molecules.

  9. A comprehensive serological and supplemental evaluation of hepatitis B "seroyield" blood donors: A cross-sectional study from a tertiary healthcare center in India

    Prashant Pandey


    Full Text Available Background: The present study addressed the interesting findings of supplemental evaluation of hepatitis B "seroyield" donors. Materials and Methods: Each blood donor sample was tested for anti-human immunodeficiency virus type I (HIV-I/HIV type II (HIV-II, HBsAg, and anti-hepatitis C virus (HCV antibody by enhanced chemiluminescence method and subjected to individual donor-nucleic acid testing (NAT for HIV-I, hepatitis B virus (HBV, and HCV. NAT test was performed using the eSAS system, Procleix Ultrio Assay, Novartis Diagnostics, CA, US. Confirmation of HBsAg was done using HBsAg Confirmatory Kit (Ortho Clinical Diagnostics, Johnson & Johnson, USA and viral load assessment was done using Cobas TaqMan real time-polymerase chain reaction (RT-PCR assay (Roche Molecular Systems, Branchburg, NJ, USA. To provide information on the stage of infection, specimens were tested for anti-HBc total (IgG + IgM, anti-HBc IgM and HBeAg. HBeAg-negative samples were tested for anti-HBe antibody. Results: A total of 60 hepatitis B seroyield donors which showed mean initial sample/cutoff of 1.6 with enhanced chemiluminescence assay were investigated further for confirmation of disease status. All 60 cases were confirmed positive with neutralization assay (VITROS HBsAg Confirmatory Kit while no target was detected on viral load assessment with RT-PCR. Sixteen donors were HBeAg positive (4 IgM anti-HBc positive and 12 IgM anti-HBc negative and 44 were IgM anti-HBc negative, anti-HBc total positive, and anti-HBe positive. Conclusion: About 7.7% of HBsAg positive and NAT nonreactive donors (nondetectable HBV DNA could be potentially infectious (HBeAg positive, whereas rest of the donors were consistent with chronic HBV infection.

  10. A comparative analysis of HIV-specific mucosal/systemic T cell immunity and avidity following rDNA/rFPV and poxvirus-poxvirus prime boost immunisations.

    Ranasinghe, Charani; Eyers, Fiona; Stambas, John; Boyle, David B; Ramshaw, Ian A; Ramsay, Alistair J


    In this study we have firstly compared a range of recombinant DNA poxvirus prime-boost immunisation strategies and shown that combined intramuscular (i.m.) 2× DNA-HIV/intranasal (i.n.) 2× FPV-HIV prime-boost immunisation can generate high-level of HIV-specific systemic (spleen) and mucosal (genito-rectal nodes, vaginal tissues and lung tissues) T cell responses and HIV-1 p24 Gag-specific serum IgG1, IgG2a and mucosal IgG, SIgA responses in vaginal secretions in BALB/c mice. Data indicate that following rDNA priming, two rFPV booster immunisations were necessary to generate good antibody and mucosal T cell immunity. This data also revealed that mucosal uptake of recombinant fowl pox (rFPV) was far superior to plasmid DNA. To further evaluate CD8+ T cell immunity, i.m. 2× DNA-HIV/i.n. 1× FPV-HIV immunisation strategy was directly compared with single shot poxvirus/poxvirus, i.n. FPV-HIV/i.m. VV-HIV immunisation. Results indicate that the latter strategy was able to generate strong sustained HIV-specific CD8+ T cells with higher avidity, broader cytokine/chemokine profiles and better protection following influenza-K(d)Gag(197-205) challenge compared to rDNA poxvirus prime-boost strategy. Our findings further substantiate the importance of vector selection/combination, order and route of delivery when designing effective vaccines for HIV-1. PMID:21352941

  11. Real-time ed end-point Polymerase Chain Reaction per la quantizzazione del DNA di Citomegalovirus: confronto tra metodi e con il test per l’antigene pp65

    Tiziano Allice


    Full Text Available Quantitave Polymerase Chain Reaction (PCR for Cytomegalovirus (CMV DNA provides highly sensitive and specific data for detecting CMV as well as monitoring the infection and determining the appropriate antiviral strategy.To determine the clinical application of a recently introduced real-time (RT PCR assay for CMV DNA quantitation in peripheral blood leukocytes (PBLs and defining its correlation with the commercial quantitative end-point (EP PCR method COBAS AMPLICOR CMV Monitor and pp65 antigen test. Sequential PBL samples (n=158 from 32 liver transplanted patients with CMV asymptomatic infection and positive for CMV DNA by EP-PCR were retrospectively analysed with RT-PCR and studied according to pp65 antigen levels. A good correlation was found between RT-PCR and pp65 antigen test (r=0.691 and between the two PCR assays (r=0.761. RT-PCR data were significantly higher in pre-emptive treated patients (those with >20 pp65+positive cells, median value: 3.8 log10 copies/500,000 PBLs than in not-treated ones (2.9 logs.According to pp65 levels of 0, 1-10, 11-20, 21-50, 51-100 and >100 positive cells/200,000 PBLs, median CMV DNA load by RT-PCR was 2.6, 3.0, 3.6, 4.0. 4.2 and 4.8, log10 copies/ 500,000 PBLs, respectively (EP-PCR CMV DNA levels: 2. 8, 2.9, 3.8, 3.7, 3.9 and 4.0 logs. For samples with >20 pp65+cells, that is above the level at which pre-emptive therapy was started, RT-PCR values were significantly higher than in groups with less than 20 pp65+cells, whereas EP-PCR values did not significantly differ and showed a slower progression rate. Dilutions of DNA from CMV AD169 strain were used to probe RT-PCR reproducibility (between and intra-assay variability < 2% and sensitivity (100% detection rate at 10 copies/reaction, 28.5% with EP-PCR. A significant improvement is coming from the introduction of RT-PCR to the study of CMV DNA dynamics in differently CMV infected patients due to a more reliable quantitation of CMV DNA for moderate and high

  12. Community risk factors for ocular Chlamydia infection in Niger: pre-treatment results from a cluster-randomized trachoma trial.

    Abdou Amza

    Full Text Available BACKGROUND: Trachoma control programs utilize mass azithromycin distributions to treat ocular Chlamydia trachomatis as part of an effort to eliminate this disease world-wide. But it remains unclear what the community-level risk factors are for infection. METHODS: This cluster-randomized, controlled trial entered 48 randomly selected communities in a 2×2 factorial design evaluating the effect of different treatment frequencies and treatment coverage levels. A pretreatment census and examination established the prevalence of risk factors for clinical trachoma and ocular chlamydia infection including years of education of household head, distance to primary water source, presence of household latrine, and facial cleanliness (ocular discharge, nasal discharge, and presence of facial flies. Univariate and multivariate associations were tested using linear regression and Bayes model averaging. FINDINGS: There were a total of 24,536 participants (4,484 children aged 0-5 years in 6,235 households in the study. Before treatment in May to July 2010, the community-level prevalence of active trachoma (TF or TI utilizing the World Health Organization [WHO] grading system was 26.0% (95% CI: 21.9% to 30.0% and the mean community-level prevalence of chlamydia infection by Amplicor PCR was 20.7% (95% CI: 16.5% to 24.9% in children aged 0-5 years. Univariate analysis showed that nasal discharge (0.29, 95% CI: 0.04 to 0.54; P = 0.03, presence of flies on the face (0.40, 95% CI: 0.17 to 0.64; P = 0.001, and years of formal education completed by the head of household (0.07, 95% CI: 0.07 to 0.13; P = 0.03 were independent risk factors for chlamydia infection. In multivariate analysis, facial flies (0.26, 95% CI: 0.02 to 0.49; P = 0.03 and years of formal education completed by the head of household (0.06, 95% CI: 0.008 to 0.11; P = 0.02 were associated risk factors for ocular chlamydial infection. INTERPRETATION: We have found that the presence

  13. Survival of infants born to HIV-positive mothers, by feeding modality, in Rakai, Uganda.

    Joseph Kagaayi

    Full Text Available BACKGROUND: Data comparing survival of formula-fed to breast-fed infants in programmatic settings are limited. We compared mortality and HIV-free of breast and formula-fed infants born to HIV-positive mothers in a program in rural, Rakai District Uganda. METHODOLOGY/PRINCIPAL FINDINGS: One hundred eighty two infants born to HIV-positive mothers were followed at one, six and twelve months postpartum. Mothers were given infant-feeding counseling and allowed to make informed choices as to whether to formula-feed or breast-feed. Eligible mothers and infants received antiretroviral therapy (ART if indicated. Mothers and their newborns received prophylaxis for prevention of mother-to-child HIV transmission (pMTCT if they were not receiving ART. Infant HIV infection was detected by PCR (Roche Amplicor 1.5 during the follow-up visits. Kaplan Meier time-to-event methods were used to compare mortality and HIV-free survival. The adjusted hazard ratio (Adjusted HR of infant HIV-free survival was estimated by Cox regression. Seventy-five infants (41% were formula-fed while 107 (59% were breast-fed. Exclusive breast-feeding was practiced by only 25% of breast-feeding women at one month postpartum. The cumulative 12-month probability of infant mortality was 18% (95% CI = 11%-29% among the formula-fed compared to 3% (95% CI = 1%-9% among the breast-fed infants (unadjusted hazard ratio (HR = 6.1(95% CI = 1.7-21.4, P-value < 0.01. There were no statistically significant differentials in HIV-free survival by feeding choice (86% in the formula-fed compared to 96% in breast-fed group (Adjusted RH = 2.8[95%CI = 0.67-11.7, P-value = 0.16] CONCLUSIONS/SIGNIFICANCE: Formula-feeding was associated with a higher risk of infant mortality than breastfeeding in this rural population. Our findings suggest that formula-feeding should be discouraged in similar African settings.

  14. Co-infection of SENV-D among chronic hepatitis C patients treated with combination therapy with high-dose interferon-alfa and ribavirin

    Chia-Yen Dai; Liang-Yen Wang; Ming-Lung Yu; Wan-Long Chuang; Wen-Yu Chang; Shinn-Cherng Chen; Li-Po Lee; Ming-Yen Hsieh; Nei-Jen Hou; Zu-Yau Lin; Ming-Yuh Hsieh


    AIM: The clinical significance of co-infection of SENV-D among patients with chronic hepatitis C (CHC) and response of both viruses to combination therapy with high-dose interferon-alfa (IFN) plus ribavirin remain uncertain and are being investigated.METHODS: Total 164 (97 males and 67 females, the mean age 48.1±11.4 years, range: 20-73 years, 128histologically proved) naive CHC patients were enrolled in this study. SENV-D DNA was tested by PCR method.Detection of serum HCV RNA was performed using a standardized automated qualitative RT-PCR assay (COBAS AMPLICOR HCV Test, version 2.0). HCV genotypes 1a,1b, 2a, 2b, and 3a were determined by using genotypespecific primers. Pretreatment HCV RNA levels were determined by using the branched DNA assay (Quantiplex HCV RNA 3.0). There are 156 patients receiving combination therapy with IFN 6 MU plus ribavirin for 24 wk and the response to therapy is determined.RESULTS: Sixty-one (37.2%) patients were positive for SENV-D DNA and had higher mean age than those who were negative (50.7±10.6 years vs46.6±11.6 years,P = 0.026). The rate of sustained viral response (SVR)for HCV and SENV-D were 67.3% (105/156) and 56.3%(27/48), respectively. By univariate analysis, the higher rate of SVR was significantly related to HCV genotype non-1b (P<0.001), younger ages (P = 0.014), lower pretreatment levels of HCV RNA (P = 0.019) and higher histological activity index (HAI) score for intralobular regeneration and focal necrosis (P = 0.037). By multivariate analyses, HCV genotype non-1b, younger age and lower pretreatment HCV RNA levels were significantly associated with HCV SVR (odds ratio (OR)/95% confidence interval (CI): 12.098/0.02-0.19, 0.936/0.890-0.998, and 3.131/1.080-9.077, respectively). The SVR of SENV-D was higher among patients clearing SENV-D than those who had viremia at the end of therapy (P = 0.04).CONCLUSION: Coexistent SENV-D infection, apparently associated with higher ages, is found in more than onethird Taiwanese

  15. Detecção do DNA de Chlamydia trachomatis em espondiloartropatias e artrite reumatóide Detection of Chlamydia trachomatis DNA in spondyloarthropathies and rheumatoid arthritis

    Rafael Navarrete Fernandez


    Full Text Available Chlamydia trachomatis é a bactéria responsável pela doença sexualmente transmissível mais prevalente no mundo. A maioria das infecções em homens e mulheres é assintomática e, quando não diagnosticada e tratada, pode causar artrite e complicações relacionadas ao aparelho reprodutor feminino. OBJETIVO: pesquisar o DNA de C. trachomatis no líquido sinovial e urina de pacientes com espondiloartropatias e artrite reumatóide (AR, avaliar a presença de anticorpos séricos IgG e IgM anti-C. trachomatis nesses dois grupos de doenças e identificar o antígeno HLA-B27 em pacientes com espondiloartropatias. MÉTODOS: a população do estudo consistiu em 15 pacientes com espondiloartropatias: nove com espondiloartropatia indiferenciada (EI e seis com artrite reativa (ARe (grupo I e 15 pacientes com AR (grupo II. O DNA clamidial foi pesquisado em amostras de líquido sinovial e urina de todos os pacientes, empregando-se a PCR (Amplicor Roche, Suíça. Os anticorpos IgG e IgM anticlamidiais foram quantificados por imunofluorescência indireta (IFI, enquanto o HLA-B27 foi tipado em 15 pacientes do grupo I por citometria de fluxo. RESULTADOS: o DNA da C. trachomatis foi evidenciado apenas em uma amostra de líquido sinovial do grupo I (6,7%, sendo o paciente portador de ARe. Em dois pacientes com AR, o DNA clamidial foi identificado na urina (13,3%. Os anticorpos IgG anticlamidiais estavam presentes em oito pacientes da população estudada, três do grupo I (20% e cinco do grupo II (33,3%. O maior título desse anticorpo (1/256 associou-se com a presença do DNA clamidial na urina de um paciente do grupo II. O anticorpo IgM não foi detectado em nenhuma amostra dos dois grupos. O antígeno HLA-B27 foi positivo em quatro indivíduos do grupo II (26,7% e sua presença relacionou-se com sacroileíte. CONCLUSÕES: os resultados deste estudo indicam que em pacientes com diagnóstico de espondiloartropatias e artrite reumatóide, com quadro articular

  16. An important role for type Ⅲ interferon(IFN-lambda) in anti-HIV activity%新型干扰素——IFN-λ抗HIV-1感染

    赵颖岚; 孙黎; 王旭; 侯炜; 霍文哲


    Objective To examine whether IFN-λ has the ability to inhibit HIV-1 infection of blood monocyte-derived macrophages and its mechanism(s). Methods Macrophages were pretreated with IFN-λ/ IFN-λ2 for 24 h before infected by HIV-1 R5 strains (Bal, Jago, and JRFL). And then the culture supernatants were detected HIV-1 reverse transcription (RT) activity and p24 protein expression by HIV-1 BT assay and ELISA. The expressions of IFN-λ receptor, CD4, CCRS, CXCR4 were evaluated by real-time PCR. Results Both IFN-λ1 and IFN-λ2, when added to macrophage cultures, inhibited HIV-1 infection and replication. This IFN-λ-mediated anti-HIV-I activity is broad, as IFN-λ could inhibit infection by both laboratory-adapted and clinical strains of HIV-1. Investigations of mechanism(s) responsible for the IFN-λ action showed that although IFN-λ had little effect on HIV-1 entry receptor CD4 and co-receptor CCR5 and CXCR4 expression, IFN-λ inhibited HIV-I infection of macrophages through connecting with IFN-λ recep-tor. Conclusion IFN-λ could inhibit HIV-I replication in macrophages. These findings indicate that IFN-λ may have a therapeutic value in the treatment of HIV-1 infection.%目的 研究干扰素λ(IFN-λ)是否对HIV-1感染人臣噬细胞有抑制作用,并对可能介导IFN-λ抗HIV-1作用的受体和辅助受体表达水平进行研究,初步探讨其抗HIV-1感染的机制.方法 HIV-1毒株感染人巨噬细胞前用IFN-λ处理细胞24 h,感染后第4天、第8天以及第12天分别检测感染细胞上清中HIV-1逆转录酶(RT)活性和p24蛋白水平,并用实时定量PCR检测细胞上IFN-λ受体、CD4、CCR5、CXCR4的表达.结果 IFN-λ对HIV-1感染人巨噬细胞具有明显抑制作用,且此作用与其剂量及作用时间呈正相关.但IFN-λ对CD4、CCR5、CXCR4的表达影响无统计学意义.结论 IFN-λ能有效抑制HIV-1感染人口噬细胞,并证实这一作用是通过其受体发挥功能的.但IFN-λ介导的抗HIV-1

  17. HIV classification using coalescent theory

    Zhang, Ming [Los Alamos National Laboratory; Letiner, Thomas K [Los Alamos National Laboratory; Korber, Bette T [Los Alamos National Laboratory


    Algorithms for subtype classification and breakpoint detection of HIV-I sequences are based on a classification system of HIV-l. Hence, their quality highly depend on this system. Due to the history of creation of the current HIV-I nomenclature, the current one contains inconsistencies like: The phylogenetic distance between the subtype B and D is remarkably small compared with other pairs of subtypes. In fact, it is more like the distance of a pair of subsubtypes Robertson et al. (2000); Subtypes E and I do not exist any more since they were discovered to be composed of recombinants Robertson et al. (2000); It is currently discussed whether -- instead of CRF02 being a recombinant of subtype A and G -- subtype G should be designated as a circulating recombination form (CRF) nd CRF02 as a subtype Abecasis et al. (2007); There are 8 complete and over 400 partial HIV genomes in the LANL-database which belong neither to a subtype nor to a CRF (denoted by U). Moreover, the current classification system is somehow arbitrary like all complex classification systems that were created manually. To this end, it is desirable to deduce the classification system of HIV systematically by an algorithm. Of course, this problem is not restricted to HIV, but applies to all fast mutating and recombining viruses. Our work addresses the simpler subproblem to score classifications of given input sequences of some virus species (classification denotes a partition of the input sequences in several subtypes and CRFs). To this end, we reconstruct ancestral recombination graphs (ARG) of the input sequences under restrictions determined by the given classification. These restritions are imposed in order to ensure that the reconstructed ARGs do not contradict the classification under consideration. Then, we find the ARG with maximal probability by means of Markov Chain Monte Carlo methods. The probability of the most probable ARG is interpreted as a score for the classification. To our

  18. HBV and neurological impairment in HIV-infected patients

    L Manolescu


    Full Text Available Objective: HIV can affect CNS in early stages of disease and determine neurological impairment. HBV DNA was found in CSF of HIV co-infected patients, but little is known about the neurotropic character of this virus. Here we assessed the degree of association between HBV infection and neurological impairment in a large cohort of long-term survivors, HIV-infected patients that experienced multiple therapeutic schemes over time. Methods: A total of 462 HIV-1-infected patients were retrospectively followed up for 10 years for HBV infection and neurological impairment. The patients were tested for immune (flow cytometry and virological parameters of HIV infection (Roche Amplicor, version 1.5/ COBAS AmpliPrep/COBAS TaqMan HIV-1 test and for HBV infection markers (HBsAg, anti HBc: Murex Biotech ELISA tests. Many of these patients have experienced between one and six regimens such as: 2 NRTIs, 3 NRTIs, 2 NRTIs+1 NNRTI, 1 NRTI+1 NNRTI+1 PI, 2 NRTIs+2 PIs. Results: After 10 years 29.87% of the patients presented neurological impairment. Out of them 56.52% were HBV-infected. The prevalence of HIV encephalopathy (HE in our studied cohort was 22.7% and 50.4% of these patients were HBV-infected. The median HIV diagnosis age was 7 and the median age of HE diagnosis was 10. In order to establish a possible correlation between HBV infection and HE we first reviewed and excluded the main risk factors associated with HE at the moment of diagnosis: low weight, anemia, constitutional symptoms, low CD4+count, high plasma HIV-RNA load. No patient was infected with HCV. The groups of patients that presented HE and HBsAg and HE without HBsAg were balanced regarding sex, number of deceased patients, number of class C3 patients, but the patients in first group presented lower CD4 values at HE diagnosis vs patients from second group 2: 44.5 vs 95 cells/µL, p=0.3; lower nadir CD4 count: 38 vs 51 cell/µL, p=0.1; and slightly higher HIV viral load: 5.2 vs 5 log10 copies

  19. Short communication: analysis of the integrase gene from HIV type 1-positive patients living in a rural area of West Cameroon.

    Turriziani, Ombretta; Montagna, Claudia; Falasca, Francesca; Bucci, Mauro; Russo, Gianluca; Lichtner, Miriam; Sobze, Martin Sanou; Vullo, Vincenzo; Pistello, Mauro; Antonelli, Guido


    Major mutations associated with HIV-I integrase inhibitors (INI) resistance are rare in INI-naive patients. However, polymorphisms at positions that may influence the genetic barrier and/or drive the selection of specific INI resistance pathways are common in HIV non-B subtypes. The aim was to evaluate the presence of natural polymorphisms and/or INI resistance mutations in HIV-1 non-B subtype samples obtained from INI-naive patients living in rural west Cameroon. Thirty-three HIV-1 non-B samples were obtained from INI-naive African women and, as controls, 15 samples of HIV-1 subtype B were obtained from antiretroviral-naive Italian patients. The integrase gene was amplified and sequenced using Trugene Core Reagents. Several amino acid positions in B and non-B subtypes were found to be polymorphic. Interestingly, two patients infected with the CRF02_AG subtype had the resistance mutations N155H and E157Q/E and 12% of African samples had an amino acid substitution at position 143. Silent mutations leading to a higher increment of genetic barriers were detected at 140 and 151 positions in non B-subtypes. Although most polymorphisms may have little effect on INI susceptibility, the IN gene variations found in the present study should be taken into consideration as they may facilitate or delay the emergence of variants fully resistant to INIs. PMID:22214532

  20. Cocaine potentiates astrocyte toxicity mediated by human immunodeficiency virus (HIV-1 protein gp120.

    Yanjing Yang

    Full Text Available It is becoming widely accepted that psychoactive drugs, often abused by HIV-I infected individuals, can significantly alter the progression of neuropathological changes observed in HIV-associated neurodegenerative diseases (HAND. The underlying mechanisms mediating these effects however, remain poorly understood. In the current study, we explored whether the psychostimulant drug cocaine could exacerbate toxicity mediated by gp120 in rat primary astrocytes. Exposure to both cocaine and gp120 resulted in increased cell toxicity compared to cells treated with either factor alone. The combinatorial toxicity of cocaine and gp120 was accompanied by an increase in caspase-3 activation. In addition, increased apoptosis of astrocytes in the presence of both the agents was associated with a concomitant increase in the production of intracellular reactive oxygen species and loss of mitochondrial membrane potential. Signaling pathways including c-jun N-teminal kinase (JNK, p38, extracellular signal-regulated kinase (ERK/mitogen-activated protein kinases (MAPK, and nuclear factor (NF-κB were identified to be major players in cocaine and gp120-mediated apoptosis of astrocytes. Our results demonstrated that cocaine-mediated potentiation of gp120 toxicity involved regulation of oxidative stress, mitochondrial membrane potential and MAPK signaling pathways.

  1. Contrasting HIV phylogenetic relationships and V3 loop protein similarities

    Korber, B. (Los Alamos National Lab., NM (United States) Santa Fe Inst., NM (United States)); Myers, G. (Los Alamos National Lab., NM (United States))


    At least five distinct sequence subtypes of HIV-I can be identified from the major centers of the AMS pandemic. While it is too early to tell whether these subtypes are serologically or phenotypically similar or distinct in terms of properties such as pathogenicity and transmissibility, we can begin to investigate their potential for phenotypic divergence at the protein sequence level. Phylogenetic analysis of HIV DNA sequences is being widely used to examine lineages of different viral strains as they evolve and spread throughout the globe. We have identified five distinct HIV-1 subtypes (designated A-E), or clades, based on phylogenetic clustering patterns generated from genetic information from both the gag and envelope (env) genes from a spectrum of international isolates. Our initial observations concerning both HIV-1 and HIV-2 sequences indicate that conserved patterns in protein chemistry may indeed exist across distant lineages. Such patterns in V3 loop amino acid chemistry may be indicative of stable lineages or convergence within this highly variable, though functionally and immunologically critical, region. We think that there may be parallels between the apparently stable HIV-2 V3 lineage and the previously mentioned HIV-1 V3 loops which are very similar at the protein level despite being distant by cladistic analysis, and which do not possess the distinctive positively charged residues. Highly conserved V3 loop protein sequences are also encountered in SIVAGMs and CIVs (chimpanzee viral strains), which do not appear to be pathogenic in their wild-caught natural hosts.

  2. Contrasting HIV phylogenetic relationships and V3 loop protein similarities

    Korber, B. [Los Alamos National Lab., NM (United States)]|[Santa Fe Inst., NM (United States); Myers, G. [Los Alamos National Lab., NM (United States)


    At least five distinct sequence subtypes of HIV-I can be identified from the major centers of the AMS pandemic. While it is too early to tell whether these subtypes are serologically or phenotypically similar or distinct in terms of properties such as pathogenicity and transmissibility, we can begin to investigate their potential for phenotypic divergence at the protein sequence level. Phylogenetic analysis of HIV DNA sequences is being widely used to examine lineages of different viral strains as they evolve and spread throughout the globe. We have identified five distinct HIV-1 subtypes (designated A-E), or clades, based on phylogenetic clustering patterns generated from genetic information from both the gag and envelope (env) genes from a spectrum of international isolates. Our initial observations concerning both HIV-1 and HIV-2 sequences indicate that conserved patterns in protein chemistry may indeed exist across distant lineages. Such patterns in V3 loop amino acid chemistry may be indicative of stable lineages or convergence within this highly variable, though functionally and immunologically critical, region. We think that there may be parallels between the apparently stable HIV-2 V3 lineage and the previously mentioned HIV-1 V3 loops which are very similar at the protein level despite being distant by cladistic analysis, and which do not possess the distinctive positively charged residues. Highly conserved V3 loop protein sequences are also encountered in SIVAGMs and CIVs (chimpanzee viral strains), which do not appear to be pathogenic in their wild-caught natural hosts.

  3. Structure of a novel shoulder-to-shoulder p24 dimer in complex with the broad-spectrum antibody A10F9 and its implication in capsid assembly.

    Ying Gu

    Full Text Available Mature HIV-1 viral particles assemble as a fullerene configuration comprising p24 capsid hexamers, pentamers and dimers. In this paper, we report the X-ray crystal structures of the p24 protein from natural HIV-1 strain (BMJ4 in complex with Fab A10F9, which recognizes a conserved epitope in the C-terminal domain of the BMJ4 p24 protein. Our structures reveal a novel shoulder-to-shoulder p24 dimerization mode that is mediated by an S-S bridge at C177. Consistent with these structures, the shoulder-to-shoulder dimer that was obtained from the BMJ4 strain was also observed in p24 proteins from other strains by the introduction of a cysteine residue at position 177. The potential biological significance was further validated by the introduction of a C177A mutation in the BMJ4 strain, which then displays a low infectivity. Our data suggest that this novel shoulder-to-shoulder dimer interface trapped by this unique S-S bridge could represent a physiologically relevant mode of HIV-1 capsid assembly during virus maturation, although Cys residue itself may not be critical for HIV-I replication.

  4. No association between seropositivity for Hepatitis C virus and lichen planus: A case control study

    Das Arup


    Full Text Available Background: The epidemiological association of lichen planus (LP with hepatitis C virus (HCV infection has been recorded from some countries and HCV RNA3 has been isolated from lesional skin in patients with LP and chronic HCV infection. The observed geographical differences regarding HCV infection and LP could be immuno-genetically related. Aim: To determine whether HCV has a causal relationship with LP. Methods: Histopathologically proved cases of LP were subjected to antibody to HCV test by the Third Generation Enzyme Immunoassay Kit for the detection of antibody to HCV (Anti-HCV in human serum or plasma. They were routinely screened in the virology department by the reagent kit, HIVASE 1 + 2, adopting the "direct sandwich principle" for the assay to detect antibodies to HIV-1 and/or HIV-2. There were 150 age and sex matched controls (not suffering from LP and HIV-I and II negative, and negative for HCV. Results: Of the 104 patients studied only 2 patients (1.92% of generalized LP with disease duration of more than 3 months were found to be positive for antibodies to HCV. This was not a significant finding and no statistical methods, e.g. Chi square test etc. could be applied. Conclusion: Hepatitis C virus is not significant to the causation of LP in India.

  5. Diagnostic performance of line-immunoassay based algorithms for incident HIV-1 infection

    Schüpbach Jörg


    Full Text Available Abstract Background Serologic testing algorithms for recent HIV seroconversion (STARHS provide important information for HIV surveillance. We have previously demonstrated that a patient's antibody reaction pattern in a confirmatory line immunoassay (INNO-LIA™ HIV I/II Score provides information on the duration of infection, which is unaffected by clinical, immunological and viral variables. In this report we have set out to determine the diagnostic performance of Inno-Lia algorithms for identifying incident infections in patients with known duration of infection and evaluated the algorithms in annual cohorts of HIV notifications. Methods Diagnostic sensitivity was determined in 527 treatment-naive patients infected for up to 12 months. Specificity was determined in 740 patients infected for longer than 12 months. Plasma was tested by Inno-Lia and classified as either incident ( Results The 10 best algorithms had a mean raw sensitivity of 59.4% and a mean specificity of 95.1%. Adjustment for overrepresentation of patients in the first quarter year of infection further reduced the sensitivity. In the preferred model, the mean adjusted sensitivity was 37.4%. Application of the 10 best algorithms to four annual cohorts of HIV-1 notifications totalling 2'595 patients yielded a mean IIR of 0.35 in 2005/6 (baseline and of 0.45, 0.42 and 0.35 in 2008, 2009 and 2010, respectively. The increase between baseline and 2008 and the ensuing decreases were highly significant. Other adjustment models yielded different absolute IIR, although the relative changes between the cohorts were identical for all models. Conclusions The method can be used for comparing IIR in annual cohorts of HIV notifications. The use of several different algorithms in combination, each with its own sensitivity and specificity to detect incident infection, is advisable as this reduces the impact of individual imperfections stemming primarily from relatively low sensitivities and

  6. No evidence for transmission of SIVwrc from western red colobus monkeys (piliocolobus badius badius to wild west african chimpanzees (pan troglodytes verus despite high exposure through hunting

    Liegeois Florian


    Full Text Available Abstract Background Simian Immunodeficiency Viruses (SIVs are the precursors of Human Immunodeficiency Viruses (HIVs which have lead to the worldwide HIV/AIDS pandemic. By studying SIVs in wild primates we can better understand the circulation of these viruses in their natural hosts and habitat, and perhaps identify factors that influence susceptibility and transmission within and between various host species. We investigated the SIV status of wild West African chimpanzees (Pan troglodytes verus which frequently hunt and consume the western red colobus monkey (Piliocolobus badius badius, a species known to be infected to a high percentage with its specific SIV strain (SIVwrc. Results Blood and plasma samples from 32 wild chimpanzees were tested with INNO-LIA HIV I/II Score kit to detect cross-reactive antibodies to HIV antigens. Twenty-three of the samples were also tested for antibodies to 43 specific SIV and HIV lineages, including SIVwrc. Tissue samples from all but two chimpanzees were tested for SIV by PCRs using generic SIV primers that detect all known primate lentiviruses as well as primers designed to specifically detect SIVwrc. Seventeen of the chimpanzees showed varying degrees of cross-reactivity to the HIV specific antigens in the INNO-LIA test; however no sample had antibodies to SIV or HIV strain - and lineage specific antigens in the Luminex test. No SIV DNA was found in any of the samples. Conclusions We could not detect any conclusive trace of SIV infection from the red colobus monkeys in the chimpanzees, despite high exposure to this virus through frequent hunting. The results of our study raise interesting questions regarding the host-parasite relationship of SIVwrc and wild chimpanzees in their natural habitat.

  7. HBV, HCV and HIV seroprevalence among blood donors in Istanbul, Turkey: how effective are the changes in the national blood transfusion policies?

    Ali Acar


    Full Text Available The national blood transfusion policies have been changed significantly in recent years in Turkey. The purpose of this study was to determine the prevalence of HBV, HCV, and HIV in blood donors at the Red Crescent Center in Istanbul and to evaluate the effect of changes in the national blood transfusion policies on the prevalence of these infections. The screening results of 72695 blood donations at the Red Crescent Center in Istanbul between January and December 2007 were evaluated retrospectively. HBsAg, anti-HCV, and anti-HIV-1/2 were screened by microparticle enzyme immunoassay (MEIA method. Samples found to be positive for anti-HIV 1/2 and anti-HCV were confirmed by Inno-Lia HCV Ab III and Inno-Lia HIV I/II Score, respectively. The seropositivity rates for HBsAg, anti-HCV, and anti-HIV-1/2 were determined as 1.76%, 0.07%, and 0.008%, respectively. Compared to the previously published data from Red Crescent Centers in Turkey, it was found that HBV and HCV seroprevalances decreased and HIV seroprevalance increased in recent years. In conclusion, we believe that the drop in HBV and HCV prevalence rates are likely multifactorial and may have resulted from more diligent donor questioning upon screening, a higher level of public awareness on viral hepatitis as well as the expansion of HBV vaccination coverage in Turkey. Another factor to contribute to the decreased prevalence of HCV stems from the use of more sensitive confirmation testing on all reactive results, thereby eliminating a fair amount of false positive cases. Despite similar transmission routes, the increase in HIV prevalence in contrast to HBV and HCV may be linked to the increase in AIDS cases in Turkey in recent years.

  8. Differences in time of virus appearance in the blood and virus-specific immune responses in intravenous and intrarectal primary SIVmac251 infection of rhesus macaques; a pilot study

    Washington Parks Robyn


    Full Text Available Abstract Background HIV-I can be transmitted by intravenous inoculation of contaminated blood or blood product or sexually through mucosal surfaces. Here we performed a pilot study in the SIVmac251 macaque model to address whether the route of viral entry influences the kinetics of the appearance and the size of virus-specific immune in different tissue compartments. Methods For this purpose, of 2 genetically defined Mamu-A*01-positive macaques, 1 was exposed intravenously and the other intrarectally to the same SIVmac251 viral stock and virus-specific CD8+ T-cells were measured within the first 12 days of infection in the blood and at day 12 in several tissues following euthanasia. Results Virus-specific CD8+ T-cell responses to Gag, Env, and particularly Tat appeared earlier in the blood of the animal exposed by the mucosal route than in the animal exposed intravenously. The magnitude of these virus-specific responses was consistently higher in the systemic tissues and GALT of the macaque exposed by the intravenous route, suggesting a higher viral burden in the tissues as reflected by the faster appearance of virus in plasma. Differences in the ability of the virus-specific CD8+ T-cells to respond in vitro to specific peptide stimulation were also observed and the greatest proliferative ability was found in the GALT of the animal infected by the intrarectal route. Conclusions These data may suggest that the natural mucosal barrier may delay viral spreading. The consequences of this observation, if confirmed in studies with a larger number of animals, may have implications in vaccine development.

  9. Sexual Behavior Varies Between Same-Race and Different-Race Partnerships: A Daily Diary Study of Highly Sexually Active Black, Latino, and White Gay and Bisexual Men.

    Grov, Christian; Rendina, H Jonathon; Ventuneac, Ana; Parsons, Jeffrey T


    Racial homophily (partnering with those of the same race) has been suggested as contributing to racial disparities in HIV among gay and bisexual men (GBM). Using a daily diary study, we examined racial homophily and its role in anal sexual behaviors in a sample of highly sexually active Black, White, and Latino GBM (N = 294, n = 3107 sexual events). In general, (1) men tended to partner with others of the same race, (2) HIV was more prevalent among men of color, and (3) race acted independent of whether one would engage in behaviors that would put them at highest risk for transmitting HIV (i.e., no main or interaction effects for insertive condomless anal sex (CAS) among HIV-positive men, and no main or interaction effects for receptive CAS among HIV-negative men). There were some main and interactive effects observed for lower risk behaviors (receptive CAS among HIV-positive men and insertive CAS among HIV-negative). Our findings suggest that racial disparities in HIV may be due to a higher exposure frequency (i.e., the frequency with which one comes into contact with a partner where a transmission could occur). However, men were also less likely to have anal sex when having sex with someone of the same race-a finding that works against the premise of higher exposure frequency. Future researchers should examine both racial homophily as well as variation in sexual behavior based on same-race or different-race partnerships. PMID:26696407

  10. Sequence analysis and genotyping of genital Chlamydia trachomatis among patients with suspected-Neisseria gonorrhoeae%拟诊为淋病患者中泌尿生殖道沙眼衣原体基因分型及序列分析

    张娟娟; 卢次勇; 冯铁建; 赵广录; 张丽君; 王峰; 洪福昌; 蓝丽娜; 吴肖冰; 陶小华; 张春来


    目的 了解深圳市拟诊为淋病患者中泌尿生殖道沙眼衣原体的合并感染情况及其基因型分布和序列变异特点.方法 采集401例拟诊为淋病患者的泌尿生殖道分泌物样本,应用Roche Amplicor全自动核酸检测系统对样本进行淋球菌和沙眼衣原体双检,提取DNA,应用巢式聚合酶链反应(nested-PGR)扩增沙眼衣原体主要外膜蛋白基因(omp1)中的VS1~VS2片段,并对其进行序列测定,所获得的序列利用Mega4.0软件与标准参考株进行比对,分析确定其基因型及序列变异情况.结果 401例拟诊为淋病患者中淋球菌的感染率为82.3%(330/401),沙眼衣原体的感染率为24.2%(97/401),淋球菌和沙眼衣原体的合并感染率为21.7%(87/401).97份沙眼衣原体阳性样本中获得73份沙眼衣原体基因片段序列,共检出8个基因型,分别为E型(27.4%)、G/Ga型(23.3%)、D/Da型(16.4%)、F型(13.7%)、J型(11.0%)、H型(5.5%)、B和K型(各1.4%).序列分析发现3例(4.1%)菌株发生错义突变,分别为D/Da型、E型、G/Ga型;F型、H型、J型和K型序列虽多见碱基突变,但均为同义突变.结论淋病患者合并感染沙眼衣原体的比例较高,且泌尿生殖道沙眼衣原体的基因型以E、G/Ga、D/Da和F型为主.序列分析可以为泌尿生殖道沙眼衣原体的分子流行病学研究提供依据.%Objective To understand the prevalence rate of genital Chlamydia trachomatis among a population with suspected-Neisseria gonorrhoeae infection,the distribution of Chlamydia trachomatis genotypes,assess changes in omp1 sequences among patients with Neisseria gonorrhoeae and Chlamydia trachomatis coinfections.Methods Four hundred and one swabs were collected.Chlamydia trachomatis and Neisseria gonorrhoeae were detected by Roche Amplicor System.DNA were extracted from those samples and were amplified by nested PCR.PCR products were sequencing and analyzed by software Mega4.0.Results The prevalence of genital Chlamydia

  11. Treponema pallidum hemagglutination assay seroreactivity among healthy Indian donors and its association with other transfusion transmitted diseases

    Sangeeta Pahuja


    Full Text Available Background: The aim of the present study was to determine the prevalence of syphilis infection by Treponema pallidum hemagglutination assay (TPHA among blood donors in Delhi and to study their correlation with other markers of transfusion transmitted infections such as hepatitis C virus (HCV, human immunodeficiency virus (HIV and hepatitis B surface antigen (HBsAg so as to establish the utility of TPHA over and above venereal diseases research laboratory test (VDRL, not only as a marker for testing T. pallidum infection, but also as a marker of high risk behavior. Materials and Methods: This prospective study was carried out in the Regional Blood Transfusion Centre, Lady Hardinge Medical College and associated Sucheta Kriplani Hospital, New Delhi for a period of 2 years. Donated blood was screened for TPHA seroreactivity along with screening for anti HIV I and II, anti-HCV, HBsAg by third generation enzyme-linked immunosorbent assay test. A total of 8082 serum samples of blood donors were collected from healthy blood donors in our blood bank. They were classified into two groups- test group and control group based on TPHA positivity. The co-occurrence of HBsAg, HIV and HCV infection were determined in TPHA positive blood donors (test group in comparison with TPHA negative blood donors (control group. Results: We found the TPHA seroreactivity to be 4.4% in Delhi′s blood donors. Nearly 8.2% (663/8082 of the donated blood had serological evidence of infection by at least one pathogen (syphilis/HIV/hepatitis B virus/HCV and 6.63% (44/663 donors with positive serology had multiple infections (two or more. Quadruple infection was seen in one donor, triple infection was seen in three donors and double infection was seen in 40 donors. Prevalence of HIV seroreactivity was found to be statistically significant and HCV seroreactivity statistically insignificant in TPHA positive group in comparison to TPHA negative group. Discussion: In our study, the

  12. HIV evolution in early infection: selection pressures, patterns of insertion and deletion, and the impact of apobec

    Korber, Bette [Los Alamos National Laboratory; Bhattacharya, Tanmoy [Los Alamos National Laboratory; Giorgi, Elena [Los Alamos National Laboratory; Gaschen, B [Los Alamos National Laboratory; Daniels, M [Los Alamos National Laboratory


    The pattern of viral diversification in newly infected individuals provides information about the host environment and immune responses typically experienced by the newly transmitted virus. For example, sites that tend to evolve rapidly across multiple early-infection patients could be involved in enabling escape from common early immune responses, represent adaptation for rapid growth in a newly infected host, or reversion from less fit forms of the virus that were selected for immune escape in previous hosts. Here we investigated the diversification of HIV -I env coding sequences in 81 very early B SUbtype infections previously shown to have resulted from transmission or expansion of single viruses (n=78) or two closely related viruses (n=3). In these cases the sequence of the infecting virus can be estimated accurately, enabling inference of both the direction of substitutions as well as distinction between insertion and deletion events. By integrating information across multiple acutely infected hosts, we find evidence of adaptive evolution of HIV-1 envand identified a subset of codon sites that diversified more rapidly than can be explained by a model of neutral evolution. Of 24 such rapidly diversifying sites, 14 were either (i) clustered and embedded in CTL epitopes that were verified experimentally or predicted based on the individual's HLA or (ii) in a nucleotide context indicative of APOBEC mediated G-to-A substitutions, despite having excluded heavily hypermutated sequences prior to the analysis. In several cases, a rapidly evolving site was both embedded in an APOBEC motif and in a CTL epitope, suggesting that APOBEC may facilitate early immune escape. Ten rapidly diversifying sites could not be explained by CTL escape or APOBEC hypermutation, including the most frequently mutated site, in the fusion peptide of gp4l. We also examined the distribution, extent, and sequence context of insertions and deletions and provide evidence that the length

  13. HIV感染者调节性T细胞消耗可导致HIV特异CD8+细胞凋亡%Treg Cells Depletion Can Lead to HIV-specific CD8 + Cells Apoptosis in HIV Patients

    焦艳梅; 福军亮; 张政; 计云霞; 王蕊; 张彤; 陈德喜; 吴昊


    Objective To investigate whether the decrease of CD4 + CD25 + FoxP3 + cells (Treg) can lead to HIV-specific CD8 +cells apoptosis in HIV patients. Methods We characterized Tregs among 75 antiviral-na(i)ve HIV-I-infected individuals in this study.Treg and CD8 + cell activation and apoptosis were detected and its relevance was analyzed; Treg cells and CD8 + cells were simultaneously purified and then co-cultured in vitro, and HIV-specific CD8 + cell apoptosis was detected with pentamer and annexin V. Results Absolute counts of circulating Treg was inversely correlated with up-regulated activation and apoptosis of CD8 + T cells. In addition, Tregs were found to suppress HIV peptite-induced apoptosis of HIV-specific CD8 + T cells. Conclusion These findings indicate that the decrease in Tregs closely correlate with increased apoptosis of CD8 + T cells, especially with HIV specific CD8 + T cells.%目的 了解在HIV感染患者CD4+CD25+FoxP3+调节性T细胞(Treg)的降低是否能够引起HIV特异CD8+细胞过度凋亡.方法 本研究对75例没有治疗的HIV患者进行研究,检测其外周血中Treg细胞计数及CD8+细胞的活化及凋亡情况并分析其相关性;同时纯化Treg细胞及CD8+细胞,在体外进行共培养,用五聚体检测HIV特异CD8+细胞的活化情况.结果 在HIV感染患者,Treg细胞与CD8+细胞的活化及凋亡呈负相关,Treg细胞能够抑制HIV特异的CD8+细胞的凋亡.结论 HIV感染患者Treg细胞的降低可导致CD8+细胞、尤其HIV特异CD8+细胞的凋亡增加.

  14. Rapid Immuno-Chromatographic Assay for the Detection of Antibodies to HIV Compare with Elisa among Voluntary and Replacement Blood Donor of Mymensingh Medical College Hospital.

    Chakrabarty, P; Rudra, S; Hossain, M A; Begum, S A; Mirza, T T; Rudra, M


    Suitable algorithms based on a combination of two or more simple rapid HIV assays have been shown to have a diagnostic accuracy comparable to double enzyme-linked immunosorbent assay (ELISA) or double ELISA with Western Blot strategies. The aims of this study were to evaluate the performance of five simple rapid HIV assays using whole blood samples from voluntary and replacement blood donors & HIV-infected patients (positive samples from BSMMU, Dhaka). Five rapid HIV assays: Determine™ HIV-1/2 (Inverness Medical), SD Bioline HIV 1/2 3.0 (Standard Diagnostics Inc.), First Response HIV Card 1-2.0 (PMC Medical India Pvt Ltd.), HIV1/2 Stat-Pak Dipstick (Chembio Diagnostic System, Inc) and Uni-Gold™ HIV-1/2 (Biotech) were evaluated between 1st February to 30th June, 2013 using 400 whole blood samples from voluntary and replacement blood donors. All samples that were reactive on all or any of the five rapid assays and 10% of non-reactive samples were tested on a confirmatory Inno-Lia HIV I/II immunoblot assay (Immunogenetics). Only 01 sample including ten positive samples from BSMMU were confirmed HIV-1 antibody positive, while 399 were HIV negative. The sensitivity at initial testing of Determine, SD Bioline and Uni-Gold™ was 100% (95% CI; 99.1-100) while First Response and Stat-Pak had sensitivity of 99.5% (95% CI; 98.2-99.9) and 97.7% (95% CI; 95.7-98.9) respectively, which increased to 100% (95% CI; 99.1-100) on repeat testing. The initial specificity of the Uni-Gold™ assay was 100% (95% CI; 99.6-100) while specificities were 99.6% (95% CI; 99-99.9), 99.4% (95% CI; 98.8-99.7), 99.6% (95% CI; 99-99.9) and 99.8% (95% CI; 99.3-99.9) for Determine, SD Bioline, First Response and Stat-Pak assays, respectively. There was no any sample which was concordantly false positive in Uni-Gold™, Determine and SD Bioline assays. An alternative confirmatory HIV testing strategy based on initial testing on either SD Bioline or Determine assays followed by testing of reactive

  15. A clinical, epidemological, laboratorial, histological and ultrasonographical evaluation of anti-HCV EIA-2 positive blood donors Avaliação clínica, epidemiológica, laboratorial, histológica e ultrassonográfica de doadores de sangue anti-HCV EIA-2 positivos

    Fernando L. GONÇALES JR


    RIBA-2 positive subjects, in 37.5% of the indeterminate RIBA-2 donors and in 9% of the negative RIBA-2 donors. Chronic hepatitis has also been observed in 50% of the histopathological exams of the anti-HCV EIA-2 reagent donors which were indeterminate RIBA-2. Among 18 blood donors with minimal changes histopathological exam 11 (61% were HCV-RNA positive. Our blood donors anti-HCV reagent generally had clinical, laboratorial and histopathological features observed in patients with chronic HCV hepatitis and a high proportion could be identified in interviews and medical evaluation realized in blood blanks. Generally, these HCV infected donors are identified and discharged only by the serological tests results.Entre 1992 e 1997 foram avaliados, ambulatorialmente, 790 doadores de sangue com teste anti-HCV EIA-2 fortemente reagente (relação entre a densidade ótica da amostra / "cut-off" > 3, que haviam sido detectados na triagem sorológica do banco de sangue. Todos eram negativos para doença de Chagas, sífilis, hepatite B (HBsAg e AIDS. Amostras de sangue foram coletadas, na primeira consulta ambulatorial, para a realização de hemograma, exames bioquímicos e novos testes sorológicos para a HVC (anti-HCV EIA-2. Em 226 doadores anti-HCV EIA-2 repetidamente reagentes, realizou-se o teste suplementar de "immunoblot" para a HVC (RIBA-2. Em 209 doadores, pesquisou-se a presença do RNA do VHC pelo teste do PCR, através de exame automatizado (HCV-AMPLICOR, ROCHE. A ultra-sonografia abdominal foi realizada em 366 doadores e a biópsia hepática em 269 concordantes. Notou-se que 95,6% eram EIA-2 repetidamente reagentes, 94% eram assintomáticos e que apenas 2% referiram icterícia pregressa. Em 47% detectou-se, pelo menos, um fator de risco para a transmissão do VHC, sendo o uso de drogas E.V. o principal deles (27,8%. A transfusão de sangue foi o segundo fator na transmissão da HVC (27,2%. Hepatomegalia foi encontrada em 54%. Esplenomegalia e sinais de hipertens

  16. In vitro anti-HIV-1 activities of Qishile, a Chinese medicine effective fraction formula%中药有效部位复方奇士乐体外抗HIV-1活性研究

    杨柳萌; 王睿睿; 张高红; 张兴杰; 陈纪军; 郑永唐


    目的 评价有效部位复方奇士乐(QSL)的体外抗HIV-1药效学.方法 通过合胞体抑制、HIV-1感染细胞保护、HIV-1 p24抗原测定等方法检测急性感染中QSL对HIV-1实验株、临床分离株、耐药株的抑制作用和对慢性感染细胞中病毒复制的影响;通过ELISA方法和荧光法分别检测了QSL体外抑制HIV-1逆转录酶和蛋白酶活性作用.结果 有效部位复方制剂QSL能有效地抑制HIV-1ⅢB诱导淋巴细胞病变、保护HIV-1ⅢB感染MT-4细胞死亡、阻断HIV-1ⅢB慢性感染H9细胞与C8166细胞间融合的作用.QSL对HIV-1实验株HIV-1ⅢB、临床分离株HIV-1KM018、耐药株HIV-174V的病毒复制也有较好的抑制作用.QSL抑制HIV活性的作用机制可能为多靶点,主要是抑制HIV逆转录酶、蛋白酶和病毒进入细胞.结论 QSL是具有较好体外抗HIV-1活性的中药有效部位复方.%Aim To evaluate the anti-HIV-1 activities of Qishile ( QSL) in vitro , a Chinese medicine effective fraction formula. Methods The inhibition of syncytia formation induced by HIV -1 was determined under microscopy. The protection of HIV-1 induced MT-4 cell lytic effects was measured by MTT assay . The level of HIV-1 p24 antigen in acute and chronic HIV-1 infection was assayed by ELISA. HIV-1 reverse transcriptase and protease activites in vitro were tested by ELISA and FRET, respectively. Results QSL markedly inhibited syncytium formation induced by HIV-1 ⅢB , protected HIV-1 ⅢB induced MT-4 cell lytic effects and blocked cell-to-cell fusion. It also showed obviously inhibitory effect on the clinical strain HIV-1KM018 ancl drug resistant strain HIV-174V replication.QSL maybe inhibited HIV-I replication through multiple targets, including reverse transcriptase , protease and virus entry. Conclusion QSL is a Chinese medicine effective fraction formula with potent anti-HIV-1 activities.

  17. Spatially Defined Oligonucleotide Arrays. Technical Report for Phase II; FINAL

    ,4,1 2). The hybridization pattern of a fluorescently labeled nucleic acid target is used to gain primary structure information of the target. This format can be applied to a broad range of nucleic acid sequence analysis problems including pathogen identification, polymorphism detection, human identification, mRNA expression monitoring and de novo sequencing. In this review, we briefly describe the method of light-directed chemid synthesis to create high-density arrays of oligonucleotide probes, the method of fluorescently labeling target nucleic acids for hybridization to the probe arrays, the detection of hybridized targets by epi-fluorescence confocal scanning and the data analysis procedures used to interpret the hybridization signals. To illustrate the use of specific high-density oligonucleotide probe arrays, we describe their application to screening the reverse transcriptase (rt) and protease (pro) genes of HIV-I for polymorphisms and drug-resistance conferring mutations

  18. Evaluation of simple rapid HIV assays and development of national rapid HIV test algorithms in Dar es Salaam, Tanzania

    Mbwana Judica


    Full Text Available Abstract Background Suitable algorithms based on a combination of two or more simple rapid HIV assays have been shown to have a diagnostic accuracy comparable to double enzyme-linked immunosorbent assay (ELISA or double ELISA with Western Blot strategies. The aims of this study were to evaluate the performance of five simple rapid HIV assays using whole blood samples from HIV-infected patients, pregnant women, voluntary counseling and testing attendees and blood donors, and to formulate an alternative confirmatory strategy based on rapid HIV testing algorithms suitable for use in Tanzania. Methods Five rapid HIV assays: Determine™ HIV-1/2 (Inverness Medical, SD Bioline HIV 1/2 3.0 (Standard Diagnostics Inc., First Response HIV Card 1–2.0 (PMC Medical India Pvt Ltd, HIV1/2 Stat-Pak Dipstick (Chembio Diagnostic System, Inc and Uni-Gold™ HIV-1/2 (Trinity Biotech were evaluated between June and September 2006 using 1433 whole blood samples from hospital patients, pregnant women, voluntary counseling and testing attendees and blood donors. All samples that were reactive on all or any of the five rapid assays and 10% of non-reactive samples were tested on a confirmatory Inno-Lia HIV I/II immunoblot assay (Immunogenetics. Results Three hundred and ninety samples were confirmed HIV-1 antibody positive, while 1043 were HIV negative. The sensitivity at initial testing of Determine, SD Bioline and Uni-Gold™ was 100% (95% CI; 99.1–100 while First Response and Stat-Pak had sensitivity of 99.5% (95% CI; 98.2–99.9 and 97.7% (95% CI; 95.7–98.9, respectively, which increased to 100% (95% CI; 99.1–100 on repeat testing. The initial specificity of the Uni-Gold™ assay was 100% (95% CI; 99.6–100 while specificities were 99.6% (95% CI; 99–99.9, 99.4% (95% CI; 98.8–99.7, 99.6% (95% CI; 99–99.9 and 99.8% (95% CI; 99.3–99.9 for Determine, SD Bioline, First Response and Stat-Pak assays, respectively. There was no any sample which was

  19. Rheumatoid Case with HCV Infection

    Bita Behnava


    Full Text Available Case Presentation:A 46-year-old woman referred to our center due to abnormality in aminotransferase level during check up. She had a history of blood transfusion 12 years ago. Anti-HCV Ab by ELISA method and HCV RNA by RT-PCR were positive. HCV RNA by Amplicor HCV monitor test counted 800,000 IU/ml and the genotype was 3a by Specific Primer-Targeted Region Core method. Laboratory evaluation revealed: Hb 11.9 mg/dl, WBC 5000 /ml, platelet count 190,000/ ml, ALT 70 IU/ml, AST 65 IU/ml, Alk phosphatase 210, PT 13 second, total protein 7.2 g/dl, albumin 4 g/dl, gama globulin 1.6 g/dl, HBsAg negative and RF positive. She had a history of symmetrical polyarthritis of small joints of upper extremities and morning stiffness for 3 years ago and had been managed as rheumatoid arthritis (RA since then. She was managed with corticosteroids and methotrexate. Are there any relations between RA disease and HCV infection?HCV-related ArthritisRheumatologic complications of HCV infection are common and include mixedcryoglobulinemia, vasculitis, Sjogren’s syndrome, arthritis and fibromyalgia(1, 2. There is a welldefined picture of arthritis associated with the presence of mixed cryoglobulinemia that consists of an intermittent mono or oligoarticular,nondestructive arthritis affecting large and mediumsize joints(1. 2% to 20% of HCV-infected patients experience arthritis and as 50% experience arthralgia(3Clinical ManifestationsHCV-related arthritis (HCVra commonly presents as rheumatoid-like, symmetrical inflammatory polyarthritis involving mainly small joints or less commonly as mono- or oligoarthritis of large joints. The joints involved in HCV-related arthritis are similar to RA(4. In about two thirds of the affected individuals, morning stiffness may be severe, resolving after more than an hour(5. Clinical picture of arthritis associated with the presence of mixed cryoglobulinemia in patients with HCV infection consists of an intermittent, mono or

  20. Establishment of a one tube touchdown nested three-monoplex PCR assay for HIV-1 early diagnosis in newborns in China%用于我国新生儿HIV-1感染早期诊断的单管降落巢式三重PCR检测方法的建立

    苏雪丽; 姚均; 蒋岩; 王临虹; 韩剑锋; 张麒; 卢红艳; 贺雄


    目的 我国面临着艾滋病母婴垂直传播的挑战.发展经济、有效的新生儿HIV-1感染早期诊断方法至关重要.本研究拟结合降落PCR、巢式PCR、多重PCR技术,以期建立一种能够在同一个PCR反应体系中同时扩增Env、Gag、Pol三个基因区的单管降落巢式三重PCR方法用于我国新生儿滤纸干血斑样本(DBS)中的HIV-1前病毒DNA检测,并对所建立的方法进行初步评价.方法 样本采自云南、新疆、广西、河南四省HIV-1阳性母亲所生婴儿.用18个DBS样本进行方法优化(感染婴儿及未感染婴儿样本分别9个).采用由重组质粒及8E5细胞制备的涵盖HIV-1各亚型的DNA稀释盘确定方法的亚型适用性及检测下限.采用134个DBS样本进行临床评价(含25个感染婴儿的64个DBS样本及30个未感染婴儿的70个DBS样本).结果 所建方法对HIV-1 A、AE、AG、B、BC、C、D、F、H亚型均可适用,检测下限为3拷贝/PCR反应体系;其临床特异度100%,阳性检出率在3月龄时接近95%、6月龄时可达到100%.结论 本研究建立的方法能够检测出我国新生儿DBS样本中各亚型的HIV-1前病毒DNA,且省时、省力、节约成本,在我国新生儿HIV-1感染的早期诊断领域具备应用潜力.%Objective HIV-1 mother-to-child transmission remains a challenge in China.The development of an efficient and affordable assay for early infant HIV-1 diagnosis is crucial.In this study,an in-house one-tube PCR assay combining touchdown PCR,nested PCR and triplex PCR techniques for Chinese infant HIV-I proviral DNA detection in dried blood spot(DBS) samples was developed and optimized to amplify Env,Gag,and Pol gene fragments in one PCR tube.Primary evaluation of the assay was made.Methods DBS from infants born to HIV-1 positive mothers in Guangxi,Henan,Xinjiang and Yunnan provinces were sampled for study.9 infected samples and 9 uninfected samples were used for optimization of the assay.An HIV-1 genome

  1. Study on effects of G_2 arrest and apoptosis in Jurkat cell by HTV-1 Vpr%HIV-1 Vpr对细胞周期的影响和致凋亡作用的研究

    刘纯; 郑力文; 郑煜煌; 周华英; 何艳; 蒋永芳; 张永红; 谌资; 刘猛; 陈霞


    can also make a good foundation for further study on gene therapy.%目的 研究人免疫缺陷病毒1型(HIV-1)的vpr基因和不同变异株对宿主细胞周期和凋亡的影响,以及其致细胞周期变化和致细胞凋亡机制的两者间的可能关系.方法 将14个带有不同变异位点的中国感染者HIV-1 upr片段分别连入pcDNA3.1(+)真核表达载体,构建重组质粒.将这些重组质粒电转染Jurkat细胞,并设立保守株vpr基因转染细胞、突变株vpr-Fs基因转染细胞、空载体转染细胞和未转染细胞作为对照.经G418选择培养及RT-PCR检测目的基因转染成功后,PI染色,流式细胞仪检测被转染细胞的细胞周期分布和细胞凋亡.结果 流式细胞仪检测上述14个带有不同变异位点的HIV-1 vpr基因片段的Jurkat细胞,发现转染保守片段HIV-1 vpr的Jurkat细胞,其细胞周期出现G_2期阻滞和细胞凋亡率明显升高,但转染vpr C端截断的vpr-Fs片段的细胞、空载体peDNA3.1(+)转染细胞和未转染的Jurkat细胞无此现象.转染了,HIV-1 vpr基因序列相对应的Vpr蛋白中含有70V、85P、86G、94G突变的片段,较vpr保守片段其致感染细胞G_2期阻滞和凋亡的能力明显下降,且Vpr蛋白的AE亚型致细胞周期阻滞和致凋亡能力较其他亚型普遍为低.初步发现vpr诱导G2期阻滞百分比越高其所致凋亡率越高.结论 HIV-I vpr基因有明显的致感染细胞G_2周期阻滞和致细胞凋亡的作用,但vpr C端截断的vpr-Fs片段无此功能.首次发现中国感染者HIV-1 vpr基因表达蛋白的70V、85P、86G、94G位点突变能使其致感染细胞G_2期阻滞和凋亡的能力下降,Vpr的AE亚型致细胞周期阻滞和凋亡能力较其他亚型普遍为低.对14个变异片段的分析显示vpr诱导G_2期阻滞的程度与其致凋亡水平可能相关,提示两者的发生机制可能有一定的关联.本研究为进一步探讨HIV-1致病机制和探索可能的基因干预措施打下基础.