Typing hepatitis C virus by polymerase chain reaction with type-specific primers: application to clinical surveys and tracing infectious sources

Abstract

Based on variation in nucleotide sequence within restricted regions in the putative C (core) gene of hepatitis C virus (HCV), four groups of HCV have been postulated in a panel of 44 HCV isolates. They were provisionally designated types I, II, III and IV. A method for typing HCV was developed, depending on the amplification of a C gene sequence by polymerase chain reaction using a universal primer (sense) and a mixture of four type-specific primers (antisense). HCV types were determined by the size of the products specific to each of them. Type II was found in HCV samples from 131 (82%) of 159 blood donors, more often than in those from 48 (60%) of 80 patients with non-A, non-B (NANB) liver disease in Japan (P< 0-01). In 11 haemophiliacs who had received imported coagulation factor concentrates, type I was found in five, as against type II in four. Double infection with two different HCV types was found in two patients with chronic NANB liver disease (types I and II; II and III) and two haemophiliacs (types I and II; I and III). HCV types were identical in mother and baby in each of two examples of perinatal transmission, and were also identical in donor and recipient in a case of accidental needle exposure.

Naive and memory T cell infiltrates in chronic hepatitis C: phenotypic changes with interferon treatment

“…Serum HCV RNA was evaluated by nested polymerase chain reaction (PCR) using two sets of oligonucleotide primers which were synthesized from the highly conserved 5 0 -non-coding region of the HCV genome [22]. HCV subtyping was performed by the method of Okamoto et al [23], and subtypes were described in accordance with proposed nomenclature [24]. To assess the efficacy of IFN, serum ALT concentrations and HCV RNA were reassayed just after and 1 year after the completion of IFN treatment.…”

Prevalence and Characterization of Hepatitis C Virus in Hemodialysis Patients.

“…The serum samples taken immediately before HDprocedure were subjected to analysis for the presence of HCV-RNA by RT-PCR using a commercially available kit (Amplicor, Roche Diagnostic Systems). In patients who were positive for HCV-RNAby RT-PCR, HCVgenotypes were determined by the method described previously by Okamoto et al (9). The genomic variations in the hypervariable region 1 (HVR1) existing in the putative E2/NS-1 domain and the core region of HCVwere analyzed by using the PCR-single strand conformation polymorphism (PCR-SSCP) assay and direct sequencing, respectively.…”

Mutations in nonstructural protein 5A gene and response to interferon in hepatitis C virus genotype 2 infection

“…All patients had histologically proven chronic active hepatitis with positive anti-HCV antibodies and serum HCV RNA by reverse-transcription polymerase chain reaction (RT-PCR) with the HCV Amplicor kit (Roche Diagnositc Systems, Basel, Switzerland), the detection limit of which was 100 copies of viral genome per milliliter of serum. 28 These patients were infected with HCV genotype 2a or 2b, as determined by a mixed-primer PCR targeted to the core region of the HCV genome, 29 which can efficiently discriminate 3 genotypes (HCV-1b, -2a, and -2b) consisting of over 99% of Japanese HCV infection. 30 They were negative for serum hepatitis B surface antigen and antinuclear antibodies, and had no other causes of hepatitis, including alcohol.…”