Chau, P. Y.; Tsang, R S
A purified soluble Vi antigen was used in counterimmunoelectrophoresis (CIE), passive haemagglutination (HA) and solid phase radio-immunoassay (SPRIA) for detection of serum Vi antibody. Serum Vi antibody was detected in 13 of 14 chronic typhoid carriers by both CIE and HA. SPRIA studies showed that Vi antibodies in sera of these carriers were mainly of the IgG class. Successful treatment with amoxycillin in one carrier, who initially showed a positive Vi test, resulted in a reversion in 6 mo...
Rusconi, F; Rancilio, L; Assael, B M; Bonora, G; Cerri, M; Pietrogrande, M C; Razon, S; Serafini, L; Torti, G; Vaggi, D
A commercial latex agglutination (LA) kit (Wellcogen, Wellcome Diagnostics) used to detect bacterial polysaccharide antigens (Haemophilus influenzae type b and Streptococcus pneumoniae) was compared with a modified counterimmunoelectrophoresis technique and blood culture for etiologic diagnosis of presumptive bacterial pneumonia requiring hospitalization in 60 infants and children. Serum, urine and either sputum or nasopharyngeal secretions were collected during the first 5 days of therapy for antigen detection. Blood culture was positive in 6 of 52 (11.5%) of cases. Antigens were detected by counterimmunoelectrophoresis and/or LA in 13 of 60 (21.7%) serum samples, 2 of 16 (12.5%) unconcentrated urine samples, 19 of 42 (45.2%) urine samples concentrated 25-fold and 21 of 45 (46.7%) sputum or nasopharyngeal secretions. Antibiotic treatment for 5 days did not affect the antigen detection rate. Counter-immunoelectrophoresis was more sensitive than LA in serum and urine but not in sputum. However, because false positive reactions were frequently obtained with LA on nasopharyngeal secretions of an age-matched control group, this test appears unreliable.
Marier, R; Andriole, V T
We have compared continuous counterimmunoelectrophoresis (CCIE) and discontinuous counterimmunoelectrophoresis (DCIE) with immunodiffusion (ID) in the identification of Candida albicans antibody using HS antigen. Eighteen sera with a titer of 1/1 to 1/32 1/32 by CCIE were titered by CCIE and ID. Fewer were positive by CCIE and ID (13 of 18 and 9 of 18, respectively). Furthermore, the titers were lower when measured by these two methods in comparison to DCIE (4 of 18 and 3 of 18 sera had titers greather than or equal to 1/8 in comparison with 7 of 18 greater than or equal to 1/8 by DCIE). Finally, DCIE was found to be much faster than CCIE and ID. After 20 min of electrophoresis, 17 of 21 sera were positive by DCIE, whereas none were positive by CCIE. Even after 90 min, only 10 of 21 were positive by CCIE, whereas 4 of 21 were positive by ID. The increased sensitivity and speed of DCIE over CCIE and ID suggest that DCIE is preferable to these other methods in the measurement of candida antibody using HS antigen.
In order to distinguish true positive results in counterimmunoelectrophoresis from false positive ones an additive counterimmunoelectrophoresis was developed. The method was tested on selected mink serum samples as part of a routine testing for antibodies towards Aleutian disease virus on 3 million...... blood samples. The procedure of the method is, that a known positive serum sample is mixed with the patient serum to be tested. The result from a false positive sample will be one precipitin line towards virus and one nonspecific line. If the serum sample is a true positive one, the antibodies...... originating from the patient serum will be added to the antibodies in the standard positive serum giving only one precipitin line. The system is further extended by testing the serum samples towards an antigen preparation containing all the cellular components but free from virus....
J. A. N. Candeias
Full Text Available Os resultados obtidos na identificação de rotavirus, usando a técnica da contraimunoeletroforese, com 162 amostras de feses de crianças com quadros diarréicos agudos, mostram uma distribuição percentual de positividade de 72,7%, 77,7% e 66,6% em diferentes grupos etários de 6 a 8 meses até um ano de idade.The faecal samples of 162 children with acute diarrhea were tested by counterimmunoelectrophoresis in order to detect rotaviruses. In 72.7%, 77.7% and 66.6% of samples from children of different age groups, varying from 6 to 8 months to 1 year or older, a positive result was found.
Evaluacion de la tecnica de contraimmunoelectroforesis para determinar la potencia antigena de las vacunas antirrabicas Evaluation ot the counterimmunoelectrophoresis technique to determine the antigenic potency of antirrabic vaccines
paper presents the results obtained when using a counterimmunoelectrophoresis (CIE techniques to determine the antigens content in samples of 84 and 40 samples of lots of suckling mouse rabies and tissue culture rabies vaccines, respectively. The evaluation of rabies vaccines under, at, near and up to 0.3 UI by the NIH and CIE tests showed that for practical use, a CIE titer of 1:4 would match the minimum of NIH potency test requirement. Low degree of CIE test variability within our laboratory was observed, when two working reference and 7 lots of rabies vaccines from different origin and activity were tested in five independent assays. All titres fell within one double dilution what is an indication of its reproducibility. Good sensitivity to detect antigen deterioration in thermal degradation test was observed when samples of 3 lots of liquid suckling mouse rabies vaccine were held stored at 4ºC or at 37ºC for 4 weeks and assays by CIE and NIH tests. Finally, CIE test could apply by manufactures to follow the concentration procedures to determine the final end point which could match a desired antigenic value in the NIH potency test.
Witer Elena Vallejo López
Full Text Available
A comparison was made between latex particles agglutination (LPA and counterimmunoelectrophoresis (CIE in the diagnosis of 57 children with acute bacterial meningitis; reagents were utllized to detect infection by Haemophilus influenzae, Streptococcus pneumoniae and Neísseria meningitídís. Results of both tests were similar for diagnosis of H. ínfluenzae and S. pneumoniae; in contrast only 30.0% of cases due to N. meningitidis gave a positive result with LP A and none was detected with CIE.in 12 patients (21.0% LPA and CIE were the only tests that allowed a precise determination ot the etiology of the disease. The authors recommend LPA for the particular situation of limited availability of funds since it is more economic than CIE and the quality of the results is similar.Se estudiaron 57 pacientes con meningitis aguda, de etiología bacteriana comprobada; 47.4% (27 casos fueron causados por Haemophilus influenzae tipo b; 21.0% (12 casos por Streptococcus pneumoniae; 17.5% (10 casos por Neisseria meningitidis; 5.3% (3 casos por Staphylococcus aureus,. 5.3% (3 casos por enterobacterias y 3.5% (2 casos por gérmenes no Identificados por cultivos. Se comparó la aglutinación de partículas de látex (APL con la contralnmunoelectroforesis (CIE en los pacientes con cultivo positivo. La exactitud de ambas fue similar para el H. influenzae tipo b y el S. pneumoniae. Tres de los 10 casos con cultivo positivo para N. meningítidis fueron positivos en la APL pero ninguno lo fue en la CIE. Se presentó un falso positivo para H. ínfluenzae con la APL que correspondió a meningitis por Salmonella typhí, Las pruebas inmunológicas estuvieron plenamente justificadas en 12 de los 57 pacientes (21.0%, previamente tratados, en quienes la bacteriología tradicional fue negativa o se quería identificar el germen porque lo único positivo era el gram y se justificaba utilizar el
Colding, H; Lind, I
was performed with antisera to Neisseria meningitidis (groups A, B and C), Streptococcus pneumoniae (omni-serum and pools A to 1), and Haemophilus influenzae type b. Antigen was detected in 57% (72/126) of specimens in which cultures revealed these three kinds of microorganisms in CSF and in 12% (17...
Grinstein, S; López, E L; Muchinik, G; Duhart, E; Bonesana, N
The serum of 50 patients between 25 days to 15 years old who were admitted with the diagnosis of pertussis syndrome were investigated for precipitating antibodies by means of discontinuous counterimmunolectrophoresis (CIED). The presence of antibodies for Bordetella pertussis antigen was shown in 28 cases. All samples were taken within the first 24/48 hours of admission and at convalescence. The technique is easy to carry out, quick and of low cost. This test offers an adequate and fast means to differentiate a Bordetella pertussis syndrome form others produced by different viral etiologies. Besides, this serologic technique shows earlier results and of lower cost than classic techniques such as agglutination and complement fixation which are more difficult to apply.
Evaluación de la calidad de los reactivos que se utilizan en la tecnica de contrainmunoelectroforesis para la determinación de anticuerpos antirrábicos Evaluation of the quality of the reagents employed in the counterimmunoelectrophoresis technique for the determination of rabies antibodies
A. M. Díaz
Full Text Available En este estudio se comprobó que el Instituto Butantan produce antígenos y sueros indicadores que se pueden utilizar con éxito en la prueba de contrainmunoelectroforesis para titular anticuerpos antirrábicos en personas inmunizadas. No se pudieron demostrar diferencias estadísticamente significativas entre los resultados de las pruebas de estandarización realizadas en el Instituto Butantan y las pruebas de control de referencia llevadas a cabo en el Centro Panamericano de Zoonosis. Se propone que el Instituto Butantan produzca y distribuya a nivel nacional los reactivos para que los laboratorios de diagnóstico apliquen la técnica de contrainmunoelectroforesis para la determinación de anticuerpos antirrábicos.This study demonstrated that the antigens and indicator sera produced by the Butantan Institute may be employed with success in the counterimmunoelectrophoresis technique for the titration of rabies antibodies in sera from immunized individuals. No statistically significant differences were demonstrated between the results obtained in the standardization tests carried out at the Butantan Institute and the reference control tests performed at the Pan American Zoonoses Center. It is proposed that the Butantan Institute be in charge of the production and distribution of these reagents at the national level.
A importância da contra-imunoeletroforese na detecção de antígenos nucleares extraíveis para o diagnóstico de doenças reumáticas sistêmicas The importance of counterimmunoelectrophoresis in the detection of extractable nuclear antigens for the diagnosis of systemic rheumatic diseases
Rita de Cassia Siqueira Bruder
Full Text Available Os antígenos nucleares extraíveis (ENAs são encontrados no soro da maioria dos pacientes com doença reumática sistêmica. Os principais ENAs estudados são SS-A/Ro, SS-B/La, RNP, Sm, Scl-70 e Jo-1. Objeti-vou-se neste trabalho: a padronizar a técnica de contra-imunoeletroforese (CIE para a detecção de ENAs; b padronizar o substrato antigênico (ENAs para a CIE a partir de baço de cão; c comparar os resultados da CIE com as técnicas de imunofluorescência indireta (IFI e Elisa para esses antígenos. Para tal foram estudados 40 soros de pacientes com doença reumática sistêmica confirmada por exames clínico e laboratorial (sorológico e biópsia. Como controle negativo foram utilizados dez soros de doadores de sangue normais, e, como controles positivos, seis soros-padrão anti-Ro, anti-La, anti-RNP, anti-Sm, anti-Scl-70, anti-Jo-1, para caracterizar os antígenos presentes no extrato de baço. Neste último foram detectados vários ENAs, exceto RNP e Scl-70. A técnica de CIE apresentou boas sensibilidade (70% e especificidade (100% em relação às outras técnicas (IFI e Elisa. A titulação dos soros pela CIE revelou positividade até diluições de 1:16 em 32,5% dos casos. Concluímos que a CIE e os antígenos extraídos de baço de cão podem ser utilizados na rotina laboratorial para triagem destes ENAs, com a vantagem, em relação à IFI, de poderem ser titulados.Extractable nuclear antigens (ENAs are detected in the sera of the majority of patients with rheumatic systemic disease. The main ENAs studied are SS-A/Ro, SS-B/La, RNP, Sm, Scl-70 and Jo-1. The aim of this work was: a to standardize the counterimmunoelectrophoresis technique (CIE to detect ENAs; b to standardize the extration of ENAs from dog spleen as a substract for CIE test; c to compare the results obtained by CIE with those of indirect immunofluorescence (IFI and Elisa for these antigens. Forty sera from individuals with rheumatic systemic disease confirmed by
Evaluation of the counterimmunoelectrophoresis reaction as a serodiagnosis test for swine leptospirosis Avaliação da reação de contraimunoeletroforese como teste sorológico para diagnóstico da leptospirose suína
Margareth Élide Genovez
Full Text Available The Counterimmunoelectrophoresis reaction (CIE was tested as a genus specific test for swine leptospirosis diagnosis using three soluble extracts obtained from Leptospira sp, serovars pomona, icterohaemorrhagiae and patoc. The extracts were treated with hot Triton X-100 and applied to serum samples of animals divided in three groups: Group 1, 10 swines experimentally infected with the Pomona strain; Group 2, 50 naturally infected swines and Group 3, 10 swines control. These animals were serologically evaluated by CIE and by the Microscopic Agglutination Test (MAT, the WHO reference technique. Groups 1 and 3 were monitored during a period of 93 days after infection (a.i.. Group 1 serum convertion took place around the 10th day a.i. by MAT but ocurred earlier by CIE using any antigen. When CIE was carried out with the homologous antigen to the experimental infection, the results were consistent with MAT, but not when the heterologous antigens were used. Groups 1 and 3 showed distinct results: in Group 3, diferences between results of CIE accomplished with any three antigen extracts were not significant, indicating lack of dependence on the serovar responsible for the outbreak. Although being safe, fast, easy to perform, inexpensive and ideal for analysis of large number of samples, CIE revealed limited genus specificity, which is not convenient for field screening tests. The advantage of CIE is the capability to detect antibodies earlier than MAT.Avaliou-se a reação de contraimunoeletroforese (CIE como teste gênero-específico para diagnóstico da leptospirose suína, usando-se três extratos solúveis de Leptospira sp, sorovares pomona, icterohaemorrhagiae e patoc, obtidos pelo tratamento com Triton X-100 a quente e aplicados a amostras de soro de suínos subdivididos em três grupos: Grupo 1, 10 suínos experimentalmente infectados com estirpe Pomona; Grupo 2, 50 suínos naturalmente infectados e Grupo 3, controle. As amostras de soros foram
I. Beckmann (Ilse); K. de Graaff (K.); F. Meisel-Mikolajczyk; H.C.S. Wallenburg (Henk)
textabstractThe ability of counter immunoelectrophoresis (CIE) to detectBacteroides fragilis endotoxin in amniotic fluid in small concentrations was evaluated. A method was developed which, in combination with ultrafiltration, permits detection ofB. fragilis endotoxin in amniotic fluid in a concentr
Marcos I. Restrepo
Full Text Available The liver abscess is the most frequent extraintestinal complication of intestinal amoebiasis: its diagnosis is suggested by the clinical picture but it must be confirmed by paraclinic tests. Themost stringent diagnosis requires identification of E. histolytica. But this is possible only in a few cases. Serological tests greatly improve the diagnosis of this severe complication of amoebiasis. We compared the Enzyme Linfed Immunosorbent Assay and the Counterimmunoeletrophoresis techniques. Both techniques were used to detect amoebic antibodies in 50 control patients, 30 patients with liver abscess and 30 patients with intestinal amoebiasis. All the sera from control patients gave negative results iin both techniques. When analysing the sera from patients with intestinal amoebiasis, 10% of them were positive by ELISA but non by CIE. The sera of patients with liver abscess, we found that 90% were positive by the ELISA method and 66.6% by the CIE technique. In patients with amoebic liver abscess, the results showed that the ELISA was more sensitive than the CIE, as it presented a higher sensitivity (100% than that of the CIE technique (66%.O abscesso hepático é a complicação mais freqüente da amebíase intestinal: o seu diagnótico sugere-se pelo quadro clínico, mas é confirmado pelos estudos paraclínicos. Para confirmar o diagnóstico precisa-se identificar a E. histolytica, o que é apenas possível em muito poucos casos. As provas sorolôgicas melhoram notadamente o diagnóstico das complicações severas da amebíase. Em nosso estudo comparamos o teste de ELISA e a Contraimunoeletroforese (CIE. Ambas as técnicas foram utilizadas para detectar anticorpos contra ameba em 50 pacientes sem amebíase, 30 pacientes com abscesso hepático e 30 com amebíase intestinal. Todos os soros dos pacientes sem amebíase foram negativos por ambas as técnicas. Quando analisamos os soros dos pacientes com amebíase intestinal, encontrou-se que 10% destes, foram positivos para ELISA, enquanto que nenhum o foi para CIE. Nos soros dos pacientes com abscesso hepático, encontrou-se uma positividade de 90% para ELISA e 66.6% para CIE. Estes resultados mostram que o teste de ELISA foi de maior sensibilidade (100%i de sensibilidade no diagnóstico do abscesso hepático amebiano, quando comparado com a Contraimunoeletroforese (66.6%.
Marcos I. Restrepo
Full Text Available The liver abscess is the most frequent extraintestinal complication of intestinal amoebiasis: its diagnosis is suggested by the clinical picture but it must be confirmed by paraclinic tests. Themost stringent diagnosis requires identification of E. histolytica. But this is possible only in a few cases. Serological tests greatly improve the diagnosis of this severe complication of amoebiasis. We compared the Enzyme Linfed Immunosorbent Assay and the Counterimmunoeletrophoresis techniques. Both techniques were used to detect amoebic antibodies in 50 control patients, 30 patients with liver abscess and 30 patients with intestinal amoebiasis. All the sera from control patients gave negative results iin both techniques. When analysing the sera from patients with intestinal amoebiasis, 10% of them were positive by ELISA but non by CIE. The sera of patients with liver abscess, we found that 90% were positive by the ELISA method and 66.6% by the CIE technique. In patients with amoebic liver abscess, the results showed that the ELISA was more sensitive than the CIE, as it presented a higher sensitivity (100% than that of the CIE technique (66%.
COUNTERIMMUNOELECTROPHORESIS (CIE) BACTERIOLOGY MYCOLOGY PARASITOLOGY VIROLOGY HAEMOPHILUS ASPERGILLUS AMEBIASIS ARBOVIRUS INFECTIONS MENINGITIS...pneumococcal antigens. Journal of of amebiasis . American Journal of Tropical Medicine Laboratory and Clinical Medicine, 81, 770-777. and Hygiene, 23, 27
Hyperimmune antirabies sera titration by standard mouse neutralization and counterimmunoelectrophoresis tests, comparing results of different laboratories Titulação de soros anti-rábicos hiperimunes, através da soroneutralização em camundongos e contraimunoeletroforese. Comparação dos resultados obtidos em diferentes laboratórios
Ana Maria Díaz
Full Text Available To determine the rabies antibody level of twenty-four hyperimmune equine sera, Standard Mouse Neutralization (SMN and Couterimmunoelectrophoresis (CIE tests were carried out, both at the Instituto Butantan (IB and Instituto Panamericano de Protección de Alimentos y Zoonosis (INPPAZ. Statistical analysis has shown a correlation (r of 0.9317 between the SMN and CIE performed at the IB, while at the INPPAZ it scored 0.974. Comparison of CIE data of both laboratories yielded a correlation of 0.845. The CIE technique has shown to be as sensitive and efficient as the SMN in titrating antirabies hyperimmune equine sera. Based on CIE results, a simple, rapid and inexpensive technique, tilers of sera antibody can be reliably estimated in SMN test.Para a determinação do nível de anticorpos de vinte e quatro amostras de soros eqüinos hiperimunes contra a Raiva, foram utilizadas as técnicas de soroneutralização em camundongos (SN e contraimunoeletroforese (CIE. As provas foram realizadas nos Institutos Butantan (IB e Panamericano de Protección de Alimentos y Zoonosis (INPPAZ. A análise estatística demonstrou que para a SN e CIE feitas no IB, a correlação foi de r=0,9317, enquanto que no INPPAZ foi de r=0,974. Comparando-se os dados obtidos pela CIE nos dois laboratórios observou-se uma correlação de r=0,845. A técnica de CIE demonstrou ser tão sensível e eficiente quanto a SN na titulação de soros anti-rábicos hiperimunes de origem eqüina. Assim, com base nos resultados de CIE, que é uma técnica simples, rápida e econômica, pode-se estimar os títulos de anticorpos dos soros em SN.
Schweinle, J. E.
Normal human serum absorbed at 0 degrees C with pneumococcal serotype 1, 12, or 25 lost the ability to support polymorphonuclear leukocyte intracellular killing of some pneumococcal serotypes even if immunoglobulin was provided. The absorbed serum contained no organisms but had residual polysaccharide when measured by counterimmunoelectrophoresis against type-specific antisera. The influence of pneumococcal polysaccharide (PPS) on serum support of intracellular polymorphonuclear leukocyte kil...
ELISA ) to detect antibodies of the parasite have been successfully used by Witmer, Maayan, Farrer & Tanowitz (1983), as well as others. Counter...immunoelectrophoresis (CIE) has been used to detect antigen in feces (Craft & Nelson, 1982) and the ELISA has also been used for the detection of antigens in...found infected with Campylobacter sp. and 12 with parasites: five were E. histolytica and seven were G. Iamblia. The scuba divers now practice in
Persson, E.; Holmberg, K.
Uterine secretions were obtained from 110 women and analysed by counterimmunoelectrophoresis for the occurrence of precipitation reactions against Actinomyces israelii antigens. Precipitation reactions were found in secretions from seven women and a correlation was found between these reactions and long term use of plastic intrauterine devices. The precipitating components could not be proved to be immunoglobulins; neither could identity be shown with IgG precipitins in reference serum. The n...
Chen, Xiaowei; Song, Cailing; Liu, Yun; Qu, Liandong; Liu, Dafei; Zhang, Yun; Liu, Ming
For detection of Aleutian mink disease virus (AMDV) antibodies, an enzyme-linked immunosorbent assay (ELISA) was developed using the recombinant VP2332-452 protein as an antigen. Counterimmunoelectrophoresis (CIEP) was used as a reference test to compare the results of the ELISA and Western blotting (WB); the specificity and sensitivity of the VP2332-452 ELISA were 97.9% and 97.3%, respectively, which were higher than those of WB. Therefore, this VP2332-452 ELISA may be a preferable method for detecting antibodies against AMDV.
Advances continue to be made in methods for more reliable or more rapid means of detecting bacteremia and fungemia. The importance of blood sample volume and broth dilution has been established in controlled studies. New technology includes the use of resins that remove antimicrobials from blood samples, detection of radioactivity from organisms given radiolabeled substrate, use of dyes that stain microbial DNA and RNA, use of slides coated with growth media, and lysis-centrifugation for trapping microorganisms. Technology now being considered includes counterimmunoelectrophoresis, head-space gas chromatography, electrical impedance, microcalorimetry, and the use of lasers to detect pH changes and turbidity.
Janoff, E N; Craft, J C; Pickering, L K; Novotny, T; Blaser, M J; Knisley, C V; Reller, L B
Antigen detection methods may facilitate diagnosis of Giardia lamblia in stool specimens. As determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis and immunoblotting, G. lamblia cysts and trophozoites share several antigens, especially in the 65-kilodalton and 30- to 34-kilodalton regions. By using blind methods, we compared results obtained by counterimmunoelectrophoresis using cyst-immune rabbit serum and by enzyme-linked immunosorbent assay (ELISA) using trophozoite-immune rabbit serum with results obtained by microscopic examination of a preserved, concentrated, and permanently stained stool specimen. Results were similar when these three methods were used to examine 118 stool specimens from clinical microbiology laboratories (53 specimens with G. lamblia) and specimens from 239 day-care-center toddlers (39 specimens with G. lamblia). Compared with microscopy, we found, for counterimmunoelectrophoresis and ELISA, respectively: sensitivity, 88 versus 94%; specificity, 97 versus 95%; positive predictive value, 86 versus 76%; negative predictive value, 98 versus 97%; and concordance, 89%. The false-positive rate by ELISA was 24% (10 of 42) in day-care-center toddlers but only 3% (1 of 32) in healthy adults (P less than 0.04) as corroborated by microscopy. This discrepancy suggests that the ELISA may be more sensitive than microscopy, which is considered the reference standard, and that results may be dependent, in part, on the epidemiology of the infection in the study subjects.
Kolodny, G M
Research during the past year has been directed at induction of specific protein synthesis in differentiated mammalian cells. The Primer Hypothesis for the regulation of eukaryotic gene expression suggests that RNA transcription is primed by small molecular weight RNA. It predicts that albumin mRNA transcription in a 3T3 fibroblast, which ordinarily does not produce albumin, can be initiated by RNA primer present in liver cells. In experiments this past year, mouse fibroblasts were incubated with mouse liver RNA. These cells did indeed produce albumin which was detected by counterimmunoelectrophoresis. Confluent 3T3 cell cultures were incubated with mouse liver RNA and polylysine in serum free media. Counterimmunoelectrophoresis (CIEP) of the dialyzed and lyophilized media against mouse albumin antisera gave a single precipitin line indicating albumin snthesis by the fibroblasts. The synthesis of this protein immunologicaly similar to albumin required new albumin mRNA transcription since it was not synthesized in the presence of actinomycin, could be found in media which had been incubated with RNA significantly smaller in size than albumin mRNA, and was synthesized in the absence of added poly A containing RNA. These results represent the first reported demonstration that RNA can be taken up from the media by cells in culture, and can induce in those cells the production of a differentiated cell product not ordinarily synthesized by those cells, i.e, a change in the normal transcription pattern in those cells.
Full Text Available The serum samples were collected from 52 patients of acute viral hepatitis and 235 hospital staff from Kasturba Hospital for Infectious Diseases. HBsAg was detected in their sera by counter-immuno-electrophoresis (CIEP, reverse passive hemogglutination (RPHA and by micro-enzyme-linked-immunosorbent assay (ELISA technique. Among the patients, HBsAg was detected in 12 cases (23% by CIEP, in 18 cases (34% by RPHA and in 23 patients (45% by ELISA. In the hospital staff, HBsAg was detected in 4 samples (1.7% by CIEP, in 8 samples (3.5% by RPHA and in 32 samples (13.5% by ELISA. Thus ELISA was found to be the most sensitive technique in detecting HBsAg.
Alvarado, Primavera; Ostos, Ana; Franquiz, Nohelys; Roschman-González, Antonio; Zambrano, Edgar A; Mendoza, Mireya
We developed and analyzed an Enzyme-Linked Immunosorbent Assay (ELISA) in order to detect antibodies in sera from sporotrichosis patients. We used a crude antigen of Sporothrix schenckii sensu stricto, obtained from the mycelial phase of the fungi. Positive sera were analyzed by other serological techniques such as double immunodiffusion (IGG) and counterimmunoelectrophoresis (CIE). The assay was validated by using sera from patients with other pathologies such as: histoplasmosis, paracoccidioidomycosis, tuberculosis, leishmaniasis, lupus and healthy individuals as negative controls. For the Sporothrix schenckii sensu stricto antigen, we found a 100% of specificity by every technique and sensitivity higher than 98% with IDD, CIE and ELISA. Our results show a high sensitivity and specificity for the Sporothrix schenckii sensu stricto antigen, so it can be used for IDD, CIE and ELISA. The results suggest that this antigen could be used in conjunction with other conventional tests for differential diagnosis and may be useful for monitoring the disease progression and response to treatment.
Luzia Helena Queiroz da Silva
Full Text Available The detection of rabies antibodies is extremely valuable for epidemiological studies, determination of immune status in man, animals, and for the diagnosis of the disease. Several serological procedures have been described for this purpose. The present study reports a comparison between counterimmunoelectrophoresis test (CIET and mouse neutralization test (MNT in the detection of antibodies against rabies virus from 212 serum samples of vaccinated dogs. The agreement between both techniques was 79.7% and a significative association was demonstrated. The correlation coefficients between MNT and the CIET titers was determined considering 88 samples showing positive results in both techniques [CIET = 2 and MNT = 5 (0.13 IU/ml] and resulted r² = 0.7926 (p < 0.001. The performance of CIET system was technically simple, cheap and rapid, and thereby it could be useful for serological monitoring of dog vaccination campaigns as well as for individual analysis.
Santiso, Gabriela; Arechavala, Alicia; Maiolo, Elena; Balarezo Juncos, Silvia; Fernández, Marisa Liliana; Messina, Fernando; Bianchi, Mario; Negroni, Ricardo
A 43 year-old diabetic woman, who suffered chronic cough and brown expectoration, is presented in this clinical problem. X-ray exam and CT thorax scan showed a cavitary lung lesion, located at the upper field of the left lung. This lesion had 5 cm in diameter, with a thick wall and a spherical shadow inside. The diagnosis of chronic cavitary pulmonary coccidioidomycosis was based on the isolation of Coccidioides sp. from cultures of expectoration and bronchoalveolar lavage, and the detection of specific antibodies in immunodiffusion test and counterimmunoelectrophoresis with coccidiodin. Her diabetes was not well controlled. She was treated with intravenous amphotericin B and oral itraconazole, with good clinical response; after four months of treatment the patient abandoned clinical controls. We suppose that the patient presented a coccidioidal fungus ball, inside a chronic cavitary lesion due to pulmonary coccidiodomycosis. She came from an endemic zone of coccidioidomycosis in the Northwest of the Argentine Republic (Catamarca Province).
Dieste-Pérez, L; Blasco, J M; de Miguel, M J; Moriyón, I; Muñoz, P M
Swine brucellosis caused by Brucella suis biovar 2 is an emerging disease in Europe. Currently used diagnostic tests for swine brucellosis detect antibodies to the O-polysaccharide (O-PS) of Brucella smooth lipopolysaccharide (S-LPS) but their specificity is compromised by false-positive serological reactions (FPSRs) when bacteria carrying cross-reacting O-PS infect pigs. FPSRs occur throughout Europe, and the only tool available for a specific B. suis diagnosis is the intradermal test with Brucella protein extracts free of O-PS or S-LPS. Using sera of 162 sows naturally infected by B. suis biovar 2, 406 brucellosis-free sows, and 218 pigs of brucellosis-free farms affected by FPSR, we assessed the diagnostic performance of an indirect ELISA with rough LPS (thus devoid of O-PS) and of gel immunodiffusion, counterimmunoelectrophoresis, latex agglutination and indirect ELISA with O-PS free proteins in comparison with several S-LPS tests (Rose Bengal, complement fixation, gel immunodiffusion and indirect ELISA). When adjusted to 100% specificity, the sensitivity of the rough LPS ELISA was very low (30%), and adoption of other cut-offs resulted in poor specificity/sensitivity ratios. Although their specificity was 100%, the sensitivity of protein tests (ELISA, latex agglutination, counterimmunoelectrophoresis, and gel immunodiffusion) was only moderate (45, 58, 61 and 63%, respectively). Among S-LPS tests, gel immunodiffusion was the only test showing acceptable sensitivity/specificity (68 and 100%, respectively). Despite these shortcomings, and when the purpose is to screen out FPSR at herd level, gel immunodiffusion tests may offer a technically simple and practical alternative to intradermal testing.
Vidal Mônica Scarpelli Martinelli
Full Text Available Chromoblastomycosis (CBM is a chronic subcutaneous infection caused by several dematiaceous fungi. The most commonly etiological agent found in Brazil is Fonsecaea pedrosoi, which appears as thick walled, brownish colored cells with transverse and longitudinal division in the lesions, called "muriform cells". This disease is found worldwide but countries like Madagascar and Brazil have highest incidence. Diagnosis is made by clinical, direct and histopathologic examination and culture of specimens. Serological tests have been used to identify specific antibodies against Fonsecaea pedrosoi antigens, as well as immunotechniques have been used for CBM serological identification and diagnosis. In the present study double immunodiffusion (DID, counterimmunoelectrophoresis (CIE and immunoenzymatic test (ELISA have been used to evaluate humoral immune response in patients with CBM caused by F. pedrosoi. Metabolic antigen was used for immunoprecipitation tests (DID and CIE while somatic antigen for ELISA. Our results demonstrated 53% sensitivity and 96% specificity for DID, while CIE presented 68% sensitivity and 90.5% specificity. ELISA demonstrated 78% sensibility and 83% specificity. Serological tests can be a useful tool to study different aspects of CBM, such as helping differential diagnosis, when culture of the pathogenic agent is impossible.
Full Text Available Buffalopox is an emerging and re-emerging zoonotic viral infection. The authors investigated an extensive zoonotic outbreak of buffalopox involving many human cases. High morbidity and significant productivity losses were recorded among domestic buffalo in Kolhapur (Maharashtra, India, between February and March 2009. The outbreak involved a total of 4 000 buffalo from 21 villages in which over 10 000 buffalo were herded. The outbreak also involved 125 humans who were mostly animal handlers and milkers of all age groups. The disease inflicted a loss of approximately 40% in terms of reduced milk production and a decline in animal trade. Although pox lesions were observed on all parts of the body, the most severe were found on the inner ear. This led to otitis and pyrexia in most of the affected animals. Milkers developed pox-like lesions on the skin of their fingers, hands, forearms, forehead, ears and face, along with pyrexia, malaise and axillary lymphadenitis and lymphadenopathy. The causal agent, buffalopox virus, was confirmed using counter-immuno-electrophoresis, the serum neutralisation test, virus isolation and buffalopox virus-specific ankyrin repeat protein (C18L gene-based polymerase chain reaction. Considering the emergence and re-emergence of buffalopox virus in buffalo, cows and humans, not only in India but also in other buffalo rearing countries, regular monitoring of outbreaks and control measures are necessary to curb economic losses and also to reduce the public health impact of the disease.
Parry, J.V.; Gardner, S.D.
A competitive-type solid phase radioimmunoassay (RIA) was developed for the detection of antibody to bovine polyomavirus. Comparison of RIA and counter-immunoelectrophoresis (CIE) results on 273 cattle sera indicated that both techniques were detecting antibody of like specificity. Human sera from 256 blood donors, 219 people recently vaccinated against polio, rubella or rabies, 50 immunosuppressed patients and 472 people with various occupational exposure to cattle were tested for antibody to bovine polyomavirus, the foetal rhesus monkey kidney strain, (anti-FRKV) by RIA. Apart from one blood donor and one of 108 rabies vaccinees only those in close contact with cattle possessed anti-FRKV. Compared with 62 per cent seropositive in the natural hosts, cattle, 71 per cent of veterinary surgeons, 50 per cent of cattle farmers, 40 per cent of abattoir workers, 16 per cent of veterinary institute technical staff and 10 per cent of veterinary students were anti-FRKV positive. Our findings indicate that the theoretical hazard of FRKV infection from undetected contamination of current tissue culture derived vaccines may, in practice, be remote. Proposed wider use of primate kidney cells as substrates for new vaccines may increase this risk.
Paulovičová, Ema; Bujdáková, Helena; Chupáčová, Jarmila; Paulovičová, Lucia; Kertys, Pavol; Hrubiško, Martin
In vitro evaluation of specific anti-Candida albicans sera antibodies based on synthetically prepared complement receptor 3-related protein (CR3-RP) mimicking the structure of native complement receptor 3 in a cohort of 72 patients with atopy and recurrent Candida vulvovaginitis (RVC) revealed effective humoral response against Candida CR3-RP. The most significant have been IgM and IgA isotype antibodies (33 and 47% positive cases, respectively). The quantitative evaluation of anti-CR3RP isotype antibodies was confronted with results of commercial ELISA anti-C. albicans antibodies diagnostics based on C. albicans cell wall mannan and β-glucan antigens, the most significant correlation being observed with anti-CR3-RP IgM and anti-β-D-glucan IgM (r(2) = 0.624) followed by isotype IgA (r(2) = 0.381). The immunogenicity and immunoreactivity of CR3RP antigen in RVC patients' sera had been evaluated with regard to the results reached by counterimmunoelectrophoresis and heterogeneous enzyme immunoassay. Obviously, synthetically prepared CR3-RP mimicking the Candida cell-wall-derived structure moiety represents a promising immunological tool not only for Candida serodiagnostics, but also prospectively for follow-up of targeted antifungal therapy and as promising Candida vaccine candidate.
Negroni, R; Duré, R; Ortiz Nareto, A; Arechavala, A I; Maiolo, E I; Santiso, G M; Iovannitti, C; Ibarra-Camou, B; Canteros, C E
A histoplasmosis outbreak affecting 6 previously healthy Air Force cadets is herein presented. The patients suffered from fever and respiratory symptoms after having cleaned an abandoned hangar soiled with pigeons and bat droppings. They all presented fever, myalgia, tachypnea, and nonproductive cough. Chest X-ray and CT scan studies showed disseminated reticulonodular images affecting both lungs. Hiliar adenomegalies were also observed. All patients achieved a favourable outcome without antifungal treatment. Both serologic tests searching for specificic antibodies (immunodiffusion and counterimmunoelectrophoresis) and histoplasmin skin tests were positive in all cases. Five soil samples mixed with pigeons and bat droppings were collected from the hangar. Suspensions of these samples were inoculated into 20 hamsters by intraperitoneal injection; mycelial phase of H. capsulatum was isolated from liver and spleen cultures. The genetic profile of this strain was compared with 12 isolates obtained from Argentinean patients, and a great degree of homogeneity was observed (> 96% similarity). Although histoplasmosis is endemic in the wet Pampas, this is the first epidemic outbreak reported south of the 34th parallel.
LACAZ Carlos da Silva
Full Text Available A case of atypical disseminated cutaneous histoplasmosis in a five-year old, otherwise healthy child, native and resident in São Paulo metropolitan area is reported. Cutaneous lesions were clinically atypical. Histologic examination disclosed a granulomatous reaction but no fungal structures could be demonstrated by specific staining nor by immunohistochemical reaction. The fungus was isolated from biopsy material on two different occasions, confirming diagnosis of an unusual fungal infection. The fungus, originally thought to be a Sepedonium sp. due to the large sized, hyaline or brownish colored tuberculated macroconidia and to lack of dimorphism (yeast form at 37 °C produce H and M antigens, visualized by the immunodiffusion with rabbit anti-Histoplasma capsulatum hyperimmune serum. Patient?s serum sample was non reactive with H. capsulatum antigen by immunodiffusion, counterimmunoelectrophoresis and complement fixation tests, and immunoenzymatic assay failed to detect the specific circulating antigen. This serum was tested negative by double immunodiffusion when antigen obtained from one of the isolated samples was used. Both cultures were sent to Dr. Leo Kaufman, Ph.D. (Mycoses Immunodiagnostic Laboratory, CDC-Atlanta/USA, who identified them as H. capsulatum by the exoantigen and gen-probe tests. Both clinic and mycologic characteristics of the present case were atypical, suggesting the fungus isolated is an ?aberrant variant? of H. capsulatum var. capsulatum, as described by SUTTON et al. in 199719. Treatment with itraconazole 100 mg/day led to cure within 90 days
Hadlow, W J; Race, R E; Kennedy, R C
Information was sought on the comparative pathogenicity of four North American strains (isolates) of Aleutian disease virus for royal pastel (a non-Aleutian genotype) and sapphire (an Aleutian genotype) mink. The four strains (Utah-1, Ontario [Canada], Montana, and Pullman [Washington]), all of mink origin, were inoculated intraperitoneally and intranasally in serial 10-fold dilutions. As indicated by the appearance of specific antibody (counterimmunoelectrophoresis test), all strains readily infected both color phases of mink, and all strains were equally pathogenic for sapphire mink. Not all strains, however, regularly caused Aleutian disease in pastel mink. Infection of pastel mink with the Utah-1 strain invariably led to fatal disease. Infection with the Ontario strain caused fatal disease nearly as often. The Pullman strain, by contrast, almost never caused disease in infected pastel mink. The pathogenicity of the Montana strain for this color phase was between these extremes. These findings emphasize the need to distinguish between infection and disease when mink are exposed to Aleutian disease virus. The distinction has important implications for understanding the natural history of Aleutian disease virus infection in ranch mink.
Full Text Available Fasciolosis is a cosmopolitan parasitosis medical-veterinary importance caused by Fasciola hepatica, which affects sheep, goats and cattle; and it affects man accidentally causing an epidemic-endemic infection difficult to diagnose. The aim was to develop an indirect sandwich ELISA with 3 antibodies for detecting excretory-secretory antigens of Fasciola hepatica (ESFh. For the development of indirect sandwich ELISA were used, as capture antibody, mouse polyclonal antibodies anti ESFh and polyclonal antibodies rabbit anti-ESFh as detection antibody, at the concentrations of 10 and 5 µg/mL respectively. The conjugate used was mouse monoclonal anti- total immunoglobulins rabbit linked to peroxidase (1/1000. Were analized 31 sheep fecal samples, and the results were compared with those obtained by direct coproparasitological examination (DC and counterimmunoelectrophoresis (CIEP. The detection limit obtained for indirect sandwich ELISA was 100 ng/mL. The test had a 100% sensitivity, 96.6% specificity, positive and negative predictive values of 50% and 96.6% respectively, in relation to DC test. Comparing with CIEP the specificity obtained for indirect sandwich ELISA was 93.5% and a negative predictive value of 100%. We concluded that indirect sandwich ELISA designed is able to detect metabolic antigens in ovine feces samples and can be used for Fasciola hepatica diagnosis.
Full Text Available Objective: To study the incidence and the features of congenital heart block (CHB in patients with undifferentiated connective tissue disease (UCTD and primary Sjögren’s syndrome (pSS. Methods: We studied 81 pregnancies of 41 women attending the Outpatients’ Clinic of the Rheumatology Unit of University Hospital of Padova from July 1989 to March 2004. Twenty five of these (61% were affected with UCTD and 16 (39% with pSS. Serologic inclusion criteria was anti-Ro/La positivity, assessed by counterimmunoelectrophoresis and ELISA. Results: CHB was found in 2 out of the 46 (4,3% pregnancies followed by our Staff and in 2 out of the 35 (5,7% included in the retrospective part of the study. In 3 cases CHB was a 3rd degree block, causing pregnancy termination in 2. The only 2nd degree block was identified in one patient at the 22nd week of gestation and treated with dexamethasone and plasma-exchange. All of the women were positive to 52 kd and 60 kd Ro autoantibodies. CHB mothers had higher titer antibodies to 52 kd Ro protein than did the mothers with healthy infants (P = 0,026. Electrocardiographic abnormalities at birth were found in 3 out of 29 asymptomatic infants. One presented sinus bradycardia, the second abnormalities of ventricular repolarization, both regressed spontaneously, while the third ventricular extrasystoles which continue even now at 5 months. Conclusion: These results showed that in UCTD and pSS there is a higher incidence of CHB than that reported in Systemic Lupus Erythematosus. Electrocardiographic screening in all infants born to mothers with anti-Ro/La antibodies would seem an important measure to identify those with irreversible heart conduction abnormalities.
González, C M; López-Longo, F J; Samson, J; Monteagudo, I; Grau, R; Rodríguez-Mahou, M; St-Cyr, C; Lapointe, N; Carreño, L
A number of clinical and laboratory features of HIV infection are found in systemic lupus erythematosus (SLE). The objective of this study was to analyze the presence of circulating antibodies to small nuclear ribonucleoproteins (snRNP) in both diseases. Sera from 44 HIV-infected children, from 22 patients with childhood-onset SLE, and from 50 healthy children were studied. Anti-snRNP antibodies were detected by ELISA using recombinant and affinity-purified nuclear antigens, by counterimmunoelectrophoresis (CIE), and by immunoblotting using extractable nuclear antigens. Results included the detection of anti-snRNP antibodies by ELISA in 30 HIV-infected patients (68.1%) and 19 SLE patients (86.3%). These antibodies were directed against U1-RNP (61.3% and 77.2%, respectively), Sm (29.5% and 54.5%, respectively), 60 kDa Ro/SS-A (47.7% and 50%, respectively), and La/SS-B proteins (18.1% and 9%, respectively). None of the HIV-infected children and 11 SLE patients (50%) showed anti-snRNP antibodies by CIE. None of the HIV-infected patients showed anti-70 kDa U1-RNP or anti-D-Sm antibodies by immunoblotting. No differences between the two groups were noted on the presence of nonprecipitating anti-snRNP antibodies. No such reactivities were observed among the normal sera tested. The authors concluded that nonprecipitating anti-snRNP antibodies in HIV-infected children are as frequent as in childhood-onset SLE. The significance of these antibodies is not clear at present. Although polyreactive and low-affinity antibodies and a mechanism of molecular mimicry may explain these results, a specific stimulation of B cells by nuclear antigens could not be excluded.
López-Longo, F J; Rodríguez-Mahou, M; Escalona-Monge, M; González, C M; Monteagudo, I; Carreño-Pérez, L
The objective was to determine the sensitivity and specificity of an automated multiparameter line immunoassay system compared with other techniques for the identification of autoantibodies in rheumatic diseases. We studied sera from 90 patients. Anti-U1RNP, anti-Sm, anti-Ro/SS-A, anti-La/SS-B, anti-Jo 1 and anti-Scl 70 antibodies were identified by counterimmunoelectrophoresis (CIE) techniques, enzyme-linked immunosorbent assay (ELISA), immunoblotting (IB) using extracts of rabbit thymus and human placenta, and an automated multiparameter line immunoassay system (INNO-LIA ANA UPDATE K-1090) that detects nine different antibodies simultaneously (anti-U1RNP, anti-Sm, anti-Ro/SS-A, anti-La/SS-B, anti-Scl 70, anti-Jo 1, anticentromere, antihistone, and antiribosomal P protein). The line immunoassay system equaled or surpassed the other techniques in the identification of anti-Sm, anti-La/SS-B, anti-Jo 1 and anti-Scl 70 antibodies (sensitivity 100%, specificity 94-100%) and was similarly effective in the case of anti-U1RNP (sensitivity 87.5%, specificity 93.9%) and anti-Ro/SS-A (sensitivity 91.4%, specificity 87.2%) antibodies. In addition, this technique detected more 52 and 60 kD anti-Ro/SS-A sera than IB. Nine antibodies can be detected with this method at a cost of 25.38 Euros per serum sample. In five hours, 19 sera can be studied. The approximate cost of detecting these nine antibodies with an automated ELISA system would be 28.93 Euros, which allows 10 sera to be studied in four hours. In conclusion, the automated multiparameter line immunoassay system is a valid method for the detection of autoantibodies in rheumatic diseases. Its most notable advantages are automated simultaneous detection of several autoantibodies in the same serum and its lower cost compared with ELISA techniques.
Antibodies against Entamoeba histolytica in individuals with intestinal amoebiasis presenting cysts and/or trophozoites in the feces Anticorpos anti-Entamoeba histolylica em indivíduos com amebíase intestinal apresentando cistos e/ou trofozoitas nas fezes
Maria das Graças Carvalho
Full Text Available Serum samples were obtained from 154 individuals infected with Entamoeba histolytica (78 symptomatic and 76 asymptomatic. Twelve had trophozoites in the feces whereas 142 had only cysts. The sera were used to test the existence of antibodies anti-Entamoeba histolytica employing the Indirect Hemagglutination (IHA, Indirect Immunofluoresccnce (IFAT, Complement Fixation Reaction (CFR and Counterimmunoelectrophoresis (CIEP. For those individuals with trophozoites in their feces, 75.0 were positive by IHA and IFAT, 83.0 by CFR and 41.7 by CIEP. In individuals who had only cysts, positive results by the same tests were respectively, 5.6%, 12.0%, 19.0% and 5.6%. The difference in relation to the tilers of antibodies detected through IHA, IFAT, CFR and CIEP and in relation to the presence of trophozoites or cysts in the feces was significative for four immunological reactions when X², was employed (P Amostras de soros foram obtidas de 154 indivíduos comprovadamente parasitados pela Entamoeba histolytica (78 sintomáticos c 76 assintomáticos. Doze apresentavam trofozoitas nas fezes, enquanto 142 tinham apenas cistos. Os soros foram utilizados para testar a ocorrência de anticorpos anti-Entamoeba histolytica, empregando-se para tal, Reação de Hemaglutinação Indireta (HAI, Reação de Imunofluorescência Indireta (RIFI, Reação de Fixação de Complemento (RFC e Contraimunoeletroforese (CIEP. Entre os soros dos indivíduos com trofozoitas em suas fezes, 75,0% foram positivos para HAI e RIFI, 83,3% por RFC, e 41,7% por CIEP. Nos indivíduos que tinham apenas cistos, resultados positivos pelos mesmos testes foram respectivamente, 5,6%; 12,0%; 19,0% c 5,6%. A diferença cm relação aos títulos de anticorpos detectados através de HAI, RIFI, RFC e CIEP e em relação à presença de trofozoitas ou cistos nas fezes foi significativa para as quatro reações imunológicas, quando X² foi empregado (P < 0,05.
Full Text Available Objective: To investigate the reliability of the immunoblot method in the detection of serum immunoreactivity towards the B/B’ polypeptides of U small nuclear ribonucleoproteins (UsnRNP and to assess the significance of these antibodies in connective tissue disease (CTD patients. Methods: We tested the sera of 348 patients with CTD (101 SLE, 51 systemic sclerosis, 53 primary Sjogren’s syndrome, 27 poly/dermatomyositis, 15 rheumatoid arthritis and 101 overlap CTD, of 31 matched healthy subjects and 13 patients with primary Epstein-Barr virus (EBV infection with high titre IgG anti-EBV antibodies. IgG anti-UsnRNP antibodies were determined by immunoblotting on nuclear extract from Raji cells (an EBV-immortalised human B lymphoid cell line and Jurkat cells (a human T lymphoid cell line. Anti-dsDNA antibodies were detected by indirect immunofluorescence on Crithidia luciliae and anti-ENA by counterimmunoelectrophoresis. Anti-dsDNA activity and avidity were measured in SLE sera by ELISA with Scatchard analysis. Results were statistically analysed by chi-square and Mann-Whitney tests. Results: A high frequency of anti-B/B’ antibodies was found in the sera of CTD patients, confined to SLE (54.4% and overlap CTD with SLE features (55,2%. Anti-B/B’ immune reactivity was closely associated with other anti-UsnRNP specificities, gel precipitating anti-nRNP and anti-P antibodies. Nine out of 15 (60% anti-B/B’ positive/anti-ENA negative lupus sera on Raji blots were confirmed to be positive also on Jurkat blots. The sera from patients with EBV infection provided, on Raji blots, completely different band patterns from those obtained with auto-immune sera. Conclusions. The Sm B/B’ proteins are the predominant or, at least, the most frequently targeted antigens of the UsnRNP auto-immune response in SLE and “lupus-like” overlap CTD. Moreover, anti-B/B’ is diagnostically specific for CTD with SLE features. Immunoblotting on human B lymphoid cells
Hidatidose policística: relato de dois casos procedentes de Sena Madureira, Acre, na Amazônia brasileira Polycystic hydatid disease: report of two cases from the city of Sena Madureira , Acre, in Brazilian Amazon
Full Text Available Dois casos de hidatidose policística (HP são relatados, oriundos do mesmo município da região amazônica brasileira (Sena Madureira, Acre. Ambos tiveram a mesma apresentação e evolução clínica ao longo de dois anos de acompanhamento. Inicialmente os pacientes queixaram-se de dor abdominal no andar superior ou no hipocôndrio direito e apresentaram icterícia obstrutiva, febre, aumento de volume abdominal e emagrecimento. Por exame de imagem, além de esplenomegalia, cistos múltiplos e coalescentes foram detectados no fígado. Amostras de soro foram reagentes por contraimunoeletroforese. O tratamento com albendazol resultou em melhora parcial, com alívio sintomático e redução no tamanho das lesões. Este relato reforça a importância de estudos clínico-epidemiológicos da hidatidose policística na região amazônica brasileira, especialmente no município de Sena Madureira, onde outros pacientes com HP podem estar sem diagnóstico.Two cases of Polycystic hydatid disease (PH are reported from the same municipal district of the Brazilian Amazon region (Sena Madureira, Acre. Both had a similar clinical presentation and course over two years of follow-up. Initially the patients complained of pain in the right hypochondrium or upper abdomen and presented obstructive jaundice, fever, increased abdominal volume and weight loss. By image analysis, in addition to splenomegaly, multiple and coalescent cysts were detected in the liver. Serum samples were reactive by counterimmunoelectrophoresis. Treatment with albendazole resulted in partial improvement, with symptomatic relief and reduction in size of the lesions. This report stresses the importance of performing clinical-epidemiological studies of polycystic hydatid disease in the Brazilian Amazon and especially in the municipality of Sena Madureira where many other cases of PH may remain undiagnosed.
Ilka Maria Landgraf
Full Text Available Eighty purulent cerebrospinal fluid (CSF samples from patients with clinical evidence of meningitis were studied using the Directigen latex agglutination (LA kit to determine the presence of bacterial antigen in CSF. The results showed a better diagnostic performance of the LA test than bacterioscopy by Gram stain, culture and counterimmunoelectrophoresis (CIE, as far as Neisseria meningitidis groups B and C, and Haemophilus influenzae type b are concerned, and a better performance than bacterioscopy and culture considering Streptococcus pneumoniae. Comparison of the results with those of culture showed that the LA test had the highest sensitivity for the Neisseria meningitidis group C. Comparing the results with those of CIE, the highest levels of sensitivity were detected for N. meningitidis groups B and C. Regarding specificity, fair values were obtained for all organisms tested. The degree of K agreement when the LA test was compared with CIE exhibited better K indices of agreement for N. meningitidis groups B and C.Oitenta amostras purulentas de líquido cefalorraquidiano (LCR de pacientes com evidência clínica de meningite foram estudadas empregando-se Kit Directigen de aglutinação de latex (AL para demonstrar antígeno bacteriano no LCR. Os resultados mostraram que o teste de AL apresentou melhor desempenho diagnóstico do que bacterioscopia através da coloração de Gram, cultura e imunoeletroforese cruzada (IEC em relação à Neisseria meningitidis grupos B e C, e ao Haemophilus influenzae tipo b, e melhor do que coloração de Gram e cultura quando Streptococcus pneumoniae foi avaliado. A comparação dos resultados com os de cultura mostrou o maior nível de sensibilidade considerando-se N. meningitidis grupo C. Quanto à especificidade, os valores foram satisfatórios para todos os microrganismos testados. O grau de concordância K em relação à IEC exibiu melhores índices K de concordância para N. meningitidis grupos B e C.
Veldhoven, C H; Meilof, J F; Huisman, J G; Smeenk, R J
A characteristic of patients with autoimmune diseases such as Sjögren's syndrome and systemic lupus erythematosus is the presence of anti-Ro/SS-A and anti-La/SS-B autoantibodies in their circulation. In order to investigate specific autoantibody levels in the sera of these patients quantitative assays for the detection of both anti-Ro/SS-A and anti-La/SS-B reactivity were developed. Ro/SS-A (60 kDa) and La-SS-B (50 kDa) cDNAs were cloned and expressed in E. coli as non-fusion proteins. These were purified to homogeneity using two different purification protocols. With these recombinant antigens, specific enzyme-linked immunosorbent assays (ELISAs) were developed. 40 sera positive for anti-Ro/SS-A autoantibodies in counterimmunoelectrophoresis (CIE) were tested in both the Ro/SS-A and La/SS-B ELISA. Activity values reproducibly ranged from 1536 to 120,000 U in the Ro/SS-A ELISA and from 763 to 2,500,000 U in the La/SS-B ELISA. The suitability of these ELISAs as screening assays was further investigated by testing 200 sera sent to our laboratory for routine detection of autoantibodies to extractable nuclear antigen (ENA: anti-Sm, anti-RNP, anti-Ro/SS-A and anti-La/SS-B). Both ELISAs showed a high sensitivity and specificity (Ro/SS-A ELISA 85% and 94%, La/SS-B ELISA 100% and 98% respectively), when compared to the standard assays, the RNA-precipitation assay and the HeLa immunoblotting test. From these data we conclude that a quantitative analysis of both anti-Ro/SS-A and anti-La/SS-B autoantibodies is now possible using purified recombinant non-fusion proteins. For screening purposes the La/SS-B ELISA showed a great improvement in sensitivity for the detection of anti-La/SS-B activity in comparison to the La/SS-B CIE, while the Ro/SS-A ELISA almost equalled the performance of the Ro/SS-A CIE.
Avaliação da contraimunoeletroforese com antígenos dos sorovars icterohaemorrhagiae E patoc no diagnóstico sorológico da leptospirose humana Evaluation of counterimmunoeletrophoresis with the serovars icterohaemorrhagiae and patoc antigens in the diagnosis of human leptospirosis
Paulo H. Yasuda
Full Text Available Avaliou-se o desempenho da contraimunoeletroforese (CIE no diagnóstico sorológico da leptospirose humana utilizando três tipos de antígenos derivados da L. interrogans sorovar icterohaemorrhagiae e do sorovar patoc da L. biflexa. Comparou-se os resultados obtidos na CIE com a prova de referência a soroaglutinação microscópica (SAM. Soros pareados de 135 pacientes com leptospirose foram subdivididos em 4 grupos de acordo com os resultados da SAM. Como controle coletou-se sangue de 69 indivíduos sadios. A concordância entre as duas técnicas variou de 92,64 a 94,11%. Os resultados obtidos pela CIE com os antígenos do sorovar icterohaemorrhagiae foram mais favoráveis do que aqueles derivados do patoc. Ressaltam-se as características de elevada sensibilidade detectando anticorpos antileptospiras mais precocemente do que a microaglutinação. As características encontradas no presente estudo credenciam o emprego da CIE como um método útil e prático para o diagnóstico da leptospirose humana na fase aguda da doença.Counterimmunoelectrophoresis (CIE was applied on paired sera from 135 pacients with leptospirosis and on 69 sera from a control group. The sera from pacients were subdivided in 4 groups according to the results obtained by the Microscopic Agglutination Test (MAT. The first samples sera from 58 pacientes were non reagent by MAT. Six monthly samples of sera were taken from 7 patients to follow-up and to determine the level of agglutinin and precipitin antibodies present using MAT and CIE. Serovars icterohaemorrhagie and patoc were used as antigens. Three types of antigens were compared, 1 Triton-X-100 extracted; 2 heat extacted and 3 a pool of them. The CIE using icterohaemorrhagiae derivated antigens types agreed with MAT in 92.64, 92.64 and 94.11% of the leptospirosis sera. The patoc antigens types reacted with the control group in 7.24, 86.95 and 84.05% of the samples, and consequently were eliminated from the present
(immunodiffusion and counterimmunoelectrophoresis and histoplasmin skin tests were positive in all cases. Five soil samples mixed with pigeons and bat droppings were collected from the hangar. Suspensions of these samples were inoculated into 20 hamsters by intraperitoneal injection; mycelial phase of H. capsulatum was isolated from liver and spleen cultures. The genetic profile of this strain was compared with 12 isolates obtained from Argentinean patients, and a great degree of homogeneity was observed (> 96% similarity. Although histoplasmosis is endemic in the wet Pampas, this is the first epidemic outbreak reported south of the 34th parallel.
Leishmania braziliensis: isolation of carbohydrate-containing antigen and possibility of its use in the immunodiagnosis of American Cutaneous Leishmaniasis Leishmania braziliensis: isolamento de antígeno contendo carboidrato e a possibilidade de sua aplicação no imunodiagnóstico da Leishmaniose Tegumentar Americana
T. G. V. Silveira
Full Text Available Leishmania braziliensis is a causative agent of American Cutaneous Leishmaniasis (ACL. The 034-JCG strain, isolated from a patient from the northern region of Paraná State, Brazil, was cultivated in Blood Agar Base medium, lyophilized and submitted to phenol-water extraction. The extract was treated with RNase I. The carbohydrate containing-antigen (Ag-CHO was immunogenic to rabbits and showed at least a fraction with some negative charge at pH 8.2. This antigen showed cross-reactivity with the phenol-water extract of the growth medium used for the culture of promastigotes and with the surface antigens of promastigotes. Its composition is: 24.3% of total sugars, from which 11.2% of galactose, 7.5% of mannose and 5.6% of ribose. Protein content was 5.4% and phosphate 18.5%. The antigenic activity was maintained after: repeated freezing-thawing; lyophilization; heating at 100ºC for 30 minutes; treatment with RNase, trichloroacetic acid and sodium metaperiodate. The precipitin line obtained is Periodic Acid Schiff positive. The application of the Ag-CHO in counterimmunoelectrophoresis reaction for the immunodiagnosis of ACL showed 60% sensitivity, and no cross-reaction with the five sera of Chagas' disease patients tested. The use of this antigen in a more sensitive technique, with more samples of sera, may improve these results.A Leishmania braziliensis é um dos agentes causadores da Leishmaniose Tegumentar Americana (LTA. A cepa 034-JCG, isolada de paciente da região norte do estado do Paraná, Brasil, foi cultivada em meio Blood Agar Base, liofilizada e submetida a extração com fenol-água e o extrato obtido foi tratado com RNase I. O antígeno contendo carboidrato (Ag-CHO mostrou-se imunogênico para coelhos e apresentou pelo menos uma fração com poucas cargas negativas em pH 8,2. Este antígeno apresenta reação cruzada com extrato fenólico do meio de cultura usado para o cultivo de promastigotas e com antígenos de superfície de
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antiserum by ID and counterimmunoelectrophoresis showed a title of 1:32, and by complement fixation (micro-technique a title of 1:128. Using immunoelectrophoresis (IEF, the produced antiserum yielded 8 lines of precipitation (5 in the anodic pole and 3 in the cathodic one. In SDS-PAGE at 12.5% the antigen has presented a rather complex electro-phoretic profile (26 proteic subunits with a molecular weight ranging from 18 a > 100 kDa. Immunogenicity of the antigen was observed in all fractions of SDS-PAGE when the immu-noblotting against the antiserum was carried out.
showed H. capsulatum in microscopic examinations or in cultures, skin scraping was the most sensitive diagnostic method (94.7% of positive results, followed by biopsies (80% and bone marrow cultures (42.1%. Specific antibodies were detected in 45.4% of the patients with histoplasmosis, using 2 different antigens and 3 types of serologic reactions (complement fixation test, immunodiffusion and counterimmunoelectrophoresis. A total of 413 samples from patients with cryptococcosis were examined, 69% of them confirmed the diagnosis. The mycologic study of CSF was the most sensitive method of study, since it registered positive results in 89.5%, followed by blood cultures (61.2%, skin scrapings (42.9%, and urine cultures (41.7%. Polisaccharyde antigens from C. neoformans in organic fluids were detected in almost all the cases. The aim of this study is to compare all the suitable diagnostic methods which can be used in systemic mycosis associated with AIDS in order to find the most rapid way of diagnosis.