| Human Immunodeficiency I Virus (HIV-1) is the pathogen of Acquire immuno- deficiency syndrome (AIDS). According to the specialist estimates, the number of HIV-infected individuals and/or AIDS patients is increasing rapidly. HIV can be divided into HIV-1 and HIV-2, at least 11subtypes in HIV-1 main group, the prevalence of HIV-1 CRF01_AE strains is increasing ,of which HIV-1 CRF01_AE are prevalent subtypes in Shanghai. China is facing a rapid upsurge in cases of human immunodeficiency virus type 1 (HIV-1) and hepatitis Cvirus (HCV) infection due to large numbers of paid blood donors (PBD), injection drug users (IDU), and sexual partners of infected individuals.In this study,four overlapping segments of HIV-1 genome were amplified by the reverse transcription nested polymerase chain reaction (RT-PCR), cloned,sequenced, and analyzed phylogenetically and with SimPlot.186 plasma samples were collected from HIV-1 seropositive individuals infected through PBD, IDU, and sex contact, living in most severely affected provinces, Henan, Yunnan, Xingjiang, Jilin and Liaoning provinces. Samples with HCV viral load > 1000c/ml were amplified by RT-nested PCR, sequenced and phylogenetically analyzed for genotyping/subtyping of HCV。HIV-1 and HCV viral loads and CD4+T lymphocytes were measured for all the subjects. We concluded that:1. The cloning and characterization of 5 CRF01_AE,1 URF01_AE/B near full-length isolates were first reported in Shanghai. the method of HIV-l near full-length genome amplification was set up.2. The phylogeny analysis of the 5 sequences showed that they clustered with HIV-1CRF01_AE isoslate; these isolates were separated into a few distinct subgroups and were dispersed among the Thailand isolates. the intersample diversity among Shanghai isolates was significantly higher than that among theThailand isolates on the level of the full-length genome,gag,pol and env genes analysis; bootscan analysis using reference isolates from each subtype from A to J and recombination subtype HIV-1 CRF01_AE showed no new intersubtype recombination in all 5 sequences.3. Near full-length sequence analysis revealed that the virus was a mosaic HIV-1 composed of CRF01_AE and HIV-1B'subtypes by recombination. The breakpoints of recombination were distributed in both polymerase and envelope genomes, which were obviously different from the previously reported HIV-1 recombinants, CRF15_AE/B, CRF33_AE/B and CRF34_AE/B.4. HCV were identified as 1a (1.2%), 1b (39.9%), 2a (17.9%), 3a (10.4%), 3b (15.6%), 6a (1.2%), 6n (6.4%), and a newly unclassified subtype (7.5%).5. HCV 1b(69) and 2a (31)subtypes predominated in PBD in Henan, 3a(18) and 3b(26) in IDUs in Xingjiang and Yunnan, and 6 genotype/subtypes(26) in IDU in Yunnan. There were no significant differences in CD4+T cell counts among the different HCV subtypes.6. The viral load of HCV RNA in 1b (69)subtype was higher than that of non-1b (104) subtype( P<0.05), however, no significant differences in HIV-1 viral loads and CD4+T cell counts were found between 1b(69) and non-1b (104)subtype. ( P>0.05) Both HIV and HCV viral loads were lower in 2a(31) than non-2a(142) subtype ( P<0.05).7. The prevalence of HCV genotype/ subtype in HIV-1/HCV co-infected individuals is associated with geographic areas and transmission routes. HCV subtypes have no direct correlation with HIV infection disease progression.In this study, we reported the near full-length molecular cloning HIV-1 CRF01_AE,URF01_AE/B subtype strains in Shanghai. We first reported that HIV-1 CRF01_AE and URF01_AE/B near full-length sequences genetic characterization,which maybe the basis on pathogen properties,genetic diversity, vaccines, diagnostic methods and infection mechanism. To better understand the prevalence and geographic distribution of genotypes/subtypes of HCV and the relationship between HCV genotypes/subtypes and HIV infection disease progression in the HIV-1/HCV co-infected individuals living in high HIV-1 prevalent areas in China... |
PDF Full Text Request