Molecular epidemiology of hepatitis C virus among different groups of people in the province of Fujian, China.

Genotyping of hepatitis C virus (HCV) can provide valuable information for prognosis and treatment duration prediction. To explore the genetic diversity of HCV in Fujian Province, China, 112, 104 and 48 anti-HCV-positive serum samples were collected from volunteer blood donors, IDUs and patients, respectively, from Jan 2008 to Dec 2008 and were genotyped through sequence analysis, followed by phylogenetic analysis in the C/E1 and NS5B regions. Genotypes could be determined for 85.61 and 84.85 % of samples in the C/E1 and NS5B region, respectively. 6a was the most prevalent subtype, which accounted for 42.04 and 43.75 % in the C/E1 and NS5B region, respectively. Mixed infection and potential recombination were detected in this study. Kappa tests indicated that similar results were obtained by two genotyping methods targeting the C/E1 and NS5B regions. The differences in the main prevalent subtype between the three target groups suggest diversity of HCV prevalence in different populations.

Genetic characterization of three CRF01_AE full-length HIV type 1 sequences from Fujian Province, China.

BACKGROUND: One of the major characteristics of the human immunodeficiency virus type 1 (HIV-1) is its unusually high degree of genetic variability, which involves in genetic diagnosis, subtyping, vaccine design, and epidemiology. HIV-1 CRF01_AE is a main prevalent HIV-1 recombinant strain in China. In this study, three full-length CRF01_AE genomes from Fujian Province, China were cloned, sequenced, and analyzed; and the further genetic diversity defining and epidemiologic analysis were carried out. METHODS: Proviral DNA was extracted from non-cultured peripheral blood mononuclear cells, the near full-length HIV-1 genome was amplified and the PCR products were cloned into pCR-XL-TOPO vector and sequenced. 5'-long terminal repeat (LTR) and 3'-LTRs were amplified by additional independent PCR and cloned into pMD18T vector. Gene-based phylogenic tree was constructed and genetic distances were calculated by MEGA 3.1. Simplot was used for Bootscan analysis. RESULTS: The phylogeny and genetic distance analysis of the three near full-length sequences confirmed that these three samples clustered with CRF01_AE isolates, more close to Thailand CRF01_AE strain CM240, and were distantly related to African CRF01_AE strain 90CF402. Analysis of their genomic organization revealed the presence of nine potential open reading frames. There were no major deletions, rearrangements, or insertions in the three sequences, but an in-frame stop codon was found in tat gene of Fj051. LTRs of the three sequences contained a few nucleotides mutation. We did not find new mosaic recombinant in the three sequences. The V3 motif was GPGQ in all the three sequences, and there were only few amino acids differences in all three V3 loop sequences. CONCLUSION: This report reveals the background of the three full-length CRF01_AE genomes, the most dominantly circulating HIV-1 strain in Fujian Province, China. The work is essential for the design and development of an effective AIDS vaccine for the region.

[Sequence analysis of the full-length glycoprotein 120 gene of HIV-1 CRF01_AE in Fujian, China].

OBJECTIVE: To characterize full length glycoprotein 120 gene variations of 21 HIV-1 CRF01_AE isolated in Fujian, China, so as to help in the immunogenic research and vaccine design. METHODS: Twenty-one peripheral blood samples were randomly collected form 100 HIV-1CRF01_AE infected persons in Fujian 2004 approximately 2005. DNA was extracted, Nested-PCR was used to amplify the envelop protein full length gp120 gene. The PVR products underwent Sepharose electrophoresis. Genotype identification was done by BLAST program. Sequence analysis was conducted with Megalign and CLUSTAL1.83. Phylogenetic tree and genetic distance were analyzed by Neighbor-joining method and Distance program of MEGA software. The amino acid similarity analysis was done with DNASIS, and N-glycosylation site analysis was done with N-GLYCOSIDE program. RESULTS: Phylogenetic tree analysis showed that these 21 HIV-1 subtype CRF01_AE strains clustered with the AE reference strain, with an overall genetic distance of 9.5% +/- 2.5%. There were four types of V3 loop central motif: GPGQ (71.43%), GPGR (19.05%), GPGH (4.76%), and GQGQ (4.76%). Prediction of the potential use of co-receptors on the basis of the critical amino acids within V3 loop disclosed potential use of CCR4/CCR5 in 16 of the 21 sequences (76.19%), potential use of CCR4/CCR5 in 1 sequence (4.76%), and 4 samples (19.05%) failed to be predicted. Amino acid sequence analysis showed that V3 region was relatively conservative, whereas V1, V2, V4, and V5 wee more variable. N-linked glycosylation site analysis showed that most of the 21 sequences were relatively conservative. CONCLUSION: Sequences analysis show Most of the CRF01_AE virus strains in Fujian have a complicated source, and most of them are closely related to those of Southeast Asia and belong to the non-syncytium inducing (NSI) type.