Description of Hepatitis B Virus Genotypes in Selected Groups of Subjects from Paraguay and Brazil
María Teresa Maidana Giret, 3Maria Angélica Camargo Suarez, 4Raquel Valdovinos, Gloria Orué, 1Denise S. Rodrigues, 6Maria Eugenia F. Canciane and 1Celso F. H. Granato 1 Division of InfectiousDiseases, Federal University of São Paulo, São Paulo, Brazil 2 National Hospital of Itauguá, Ministry of Health of Paraguay, Asunción, Paraguay 3 Blood Bank, Federal University of São Paulo, São Paulo, Brazil 4 Haemodialysis Center, National Hospital of Itauguá, Paraguay 5 Haemodialysis Center, Emergency Medical Hospital Dr. Luiz M. Argaña, Asunción, Paraguay 6 Kidney and Hypertension HospitalOswaldo Ramos, Federal University of São Paulo, São Paulo, Brazil
Abstract: Hepatitis B virus genotyping was conducted among 11560 healthy blood donors and haemodialysis patients in Brazil and Paraguay. Genotypes A, D and F were found in Brazil and genotypes D and F in Paraguay. This is the first report showing HBV genotype diversity in individuals in Paraguay. Key words: Hepatitis B virus,genotypes, Paraguay INTRODUCTION Hepatitis B Virus Infection (HBV) is a relevant public health problem, with 2 billion people infected worldwide. Effective vaccines against HBV are available starting from 1982, however not all countries adopted those recommendations in an official way. An important epidemiological aspect of HBV is the tendency to establish closed epidemiological cycles,hence, detailed knowledge is essential to design and improve specific prevention and control strategies. However, this notion is still incomplete since the number of isolates analyzed in some parts of the world, including South America, is small[3,4]. HBV strains are classified into eight main genomic groups, designated A-H[5,6] showing a global distribution. In Brazil the most frequent genotypeswere A, D and F. The objective of this study was to detect and characterize HBV strains circulating among selected populations in Brazil and Paraguay, including healthy blood donors and haemodialysis patients. HBV viral diversity in Paraguay is evaluated here for the first time.
MATERIALS AND METHODS A total of 11560 serum samples from healthy blood donors andhaemodialysis patients in Brazil and Paraguay were divided into four groups. Characteristics and provenance of studied population groups are shown (Table 1). Samples were tested for HBsAg by Enzyme immunoassay (MONOLISA® HBsAg PLUS, BIORAD) and “indeterminate” samples were also registered. The haemodialysis groups were also tested for anti-HBc by Enzyme immunoassay (MONOLISA® Anti HBc PLUS, BIO-RAD).Nucleic acid was extracted from serum as previously described. Detection of HBV DNA and genotyping was carried out in the HBsAg positive samples, in the “indeterminate” samples and the HBsAg- / anti HBc+ samples; the methodology used was nested PCR using type-specific primers for each genotype, originally reported by Naito et al. with slight modifications. We amplified the HBV genome using theuniversal primers followed by two different amplifications containing type-specific inner primers designed on the basis of the differences in size of the genotype-specific bands. PCR was carried out in a tube containing 50 µl of a reaction mix with the following components: 50 ng of each outer primer, 200 mm of dNTPs, 1 U of Taq
Celso F.H. Granato, M.D., Ph.D., Laboratório de Imuno-Virologia,Disciplina de Doenças Infecciosas e Parasitárias, Escola Paulista de Medicina/ UNIFESP, Rua Pedro de Toledo, 781, 15o andar, 04039-032 - São Paulo – SP, Brazil
Am. J. Infect. Dis., 4 (4): 220-223, 2008
Table 1: Groups Group 1 Group 2 Group 3 Group 4 Total a Hd Py: BS Py: BS Br: Hd Br: b HBV serology and DNA detection in studied population groups Identify. Hd Py Bs Py Bs Br Hd Br N...