Diagnosis of hepatitis C virus in Brazilian blood donors using a reverse transcriptase nested polymerase chain reaction: comparison with enzyme immunoassay and recombinant protein immunoblot assay
Rev. Inst. Med. Trop. Säo Paulo
;
42(5): 263-67, Sept.-Oct. 2000. tab
Artículo
en Inglés
| LILACS
| ID: lil-270227
RESUMO
Screening blood donations for anti-HCV antibodies and alanine aminotransferase (ALT) serum levels generally prevents the transmission of hepatitis C virus (HCV) by transfusion. The aim of the present study was to evaluate the efficiency of the enzyme immunoassay (EIA) screening policy in identifying potentially infectious blood donors capable to transmit hepatitis C through blood transfusion. We have used a reverse transcriptase (RT)-nested polymerase chain reaction (PCR) to investigate the presence of HCV-RNA in blood donors. The prevalence of HCV-RNA positive individuals was compared with the recombinant immunoblot assay (RIBA-2) results in order to assess the usefulness of both tests as confirmatory assays. Both tests results were also compared with the EIA-2 OD/C ratio (optical densities of the samples divided by the cut off value). ALT results were expressed as the ALT quotient (qALT), calculated dividing the ALT value of the samples by the maximum normal value (53UI/l) for the method. Donors (n=178) were divided into five groups according to their EIA anti-HCV status and qALT group A (EIA > or = 3, ALT<1), group B (EIA > or = 3, ALT>1), group C (1<=EIA<3, ALT<1), group D (1<=EIA<3, ALT>1) and group E (EIA<=0.7). HCV sequences were detected by RT-nested PCR, using primers for the most conserved region of viral genome. RIBA-2 was applied to the same samples. In group A (n=6), all samples were positive by RT-nested PCR and RIBA-2. Among 124 samples in group B, 120 (96.8 percent) were RIBA-2 positive and 4 (3.2 percent) were RIBA-2 indeterminate but were seropositive for antigen c22.3. In group B, 109 (87.9 percent) of the RIBA-2 positive samples were also RT-nested PCR positive, as well as were all RIBA-2 indeterminate samples. In group C, all samples (n=9) were RT-nested PCR negative 4 (44.4 percent) were also RIBA-2 negative, 4 (44.4 percent) were RIBA-2 positive and 1 (11.1 percent) was RIBA-2 indeterminate. HCV-RNA was detected by RT-nested PCR in 3 (37.5 percent) out of 8 samples in group D. Only one of them was also RIBA-2 positive, all the others were RIBA-2 indeterminate. All of the group E samples (controls) were RT- nested PCR and RIBA-2 negative. Our study suggests a strong relation between anti-HCV EIA-2 ratio > or = 3 and detectable HCV-RNA by RT-nested PCR. We have also noted that blood donors with RIBA-2 indeterminate presented a high degree of detectable HCV-RNA using RT-nested PCR...
Texto completo:
Disponible
Índice:
LILACS (Américas)
Asunto principal:
Immunoblotting
/
Técnicas para Inmunoenzimas
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Hepatitis C
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Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
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Alanina Transaminasa
Tipo de estudio:
Estudio diagnóstico
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Estudio observacional
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Estudio de prevalencia
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Estudio pronóstico
/
Factores de riesgo
Límite:
Humanos
País/Región como asunto:
America del Sur
/
Brasil
Idioma:
Inglés
Revista:
Rev. Inst. Med. Trop. Säo Paulo
Asunto de la revista:
Medicina Tropical
Año:
2000
Tipo del documento:
Artículo
País de afiliación:
Brasil
Institución/País de afiliación:
Universidade Estadual de Campinas/BR
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