ABSTRACT
A new restriction fragment length polymorphism (RFLP) analysis has been developed for hepatitis C virus (HCV) typing in the viral 5' non-coding region and contiguous core region. These genomic sequences were chosen for the relative nucleotide homology among different genotypes and for the presence of polymorphic sites. By employing two endonucleases (AccI and MboI) and, in some instances, a third one (EcoRII), we can unambiguously and reproducibly distinguish between genotypes and subtypes 1a, 1b, 1c, 2a, 2c, 2b, 3a, 3b, 4a, 5a and 6a. The method was applied for diagnosing two Italian groups of HCV-infected individuals reflecting a randomly collected population and a group of intravenous drug users. The accuracy of this method has been validated by comparison with INNOLiPA and by sequencing. Our approach represents an improvement over previous RFLP methods, since typing is accurate and simpler.
Subject(s)
Hepacivirus/genetics , Polymorphism, Restriction Fragment Length , Base Sequence , Female , Genotype , Hepacivirus/classification , Hepatitis C/virology , Humans , Male , Molecular Sequence Data , RNA, Viral/geneticsABSTRACT
A selective rubella immunization program was carried out in the Siena (Italy) area since 1984. 912 schoolgirls aged 10 to 18 were screened for rubella antibodies. Seropositivity rates, increasing with age, appeared quite heterogenous among subjects living in different villages. Seronegative girls were partly voluntarily immunized with RA 27/3 live attenuated rubella vaccine. Serological and clinical controls performed on 238 subjects one month after immunization showed a 99.6% vaccine-induced seroconversion rate and mild post-vaccinal reactions in 26% of vaccinees. Serological control performed, on 172 subjects, at 8-12 days after vaccination revealed that in most cases vaccine induced haemagglutination-inhibiting antibodies are not detectable at the time. The results are discussed in relation to a forthcoming introduction of compulsory rubella vaccination in our Country.
