Use of conserved sequences from hepatitis C virus for the detection of viral RNA in infected sera by polymerase chain reaction

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Abstract

Three oligonucleotide primer combinations selected from the 5′ noncoding, the nucleocapsid and the putative nonstructural regions of the hepatitis C virus genome were compared in a nested polymerase chain reaction assay with respect to sensitivity and specificity for the detection of viral RNA in chimpanzeeinfected and human-infected sera. Sera from both the acute and the chronic phase of the infection were obtained from 13 animals inoculated with five different non-A, non-B hepatitis strains and from seven cardiac surgery patients who had non-A, non-B hepatitis develop after transfusion and who had been tested in parallel for the presence of hepatitis C virus RNA and anti-C100-3. A total of 90% of the acute-phase and 100% of the chronic-phase sera tested positive for hepatitis C virus RNA when the 5′ noncoding–derived or the nucleocapsid-derived combinations were used; only 58% and 56%, respectively, gave positive results with the putative nonstructural primers, whereas 33% and 71%, respectively, scored positive for C100-3. Thus polymerase chain reaction primers selected from either the highly conserved 5′ noncoding or nucleocapsid-regions appear to provide the sensitivity and the specificity necessary to detect low levels of hepatitis C virus RNA in both chimpanzee-infected and human-infected sera. (HEPATOLOGY 1991;14:595–600.)

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