The analyses of HIV-1 quasispecies in HIV-infected subjects by the application of single genome amplification assay
10.3760/cma.j.issn.0253-9624.2014.09.015
- VernacularTitle:应用单基因组扩增法分析HIV-1准种基因变异
- Author:
Yanmin QIN
1
;
Xiaohui WANG
;
Dongfeng KONG
;
Shunxiang ZHANG
Author Information
1. 518055,深圳市疾病预防控制中心传染病防制科
- Keywords:
HIV;
Gene amplification;
Quasispecies;
Limiting dilution method
- From:
Chinese Journal of Preventive Medicine
2014;(9):819-822
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish the single genome amplification ( SGA) method and analyze the quasispecies in HIV-infected patients.Methods All 6 sample RNA acquired in 2010 in Shenzhen and genetic sequenced as overlap peaks were extracted and diluted to a single copy , nest-PCR after one step RT-PCR was employed to amplify HIV-1 genome , and then PCR products was purified and sequenced.Mega 4.02 software was used to analyze the genetic distance among HIV-1 quasispecies , and phylogenetic tree was constructed.Results Our data showed that viral sequences derived from different patients were grouped into different clusters.Subcluster was also observed in several clusters , indicating these existed competition and preferential replication of certain viral strains.The genetic distance within one cluster of 6 samples were between 0.008 and 0.06, it was likely to associate with the duration since infection and viral load.Conclusion SGA is a useful approach to gain information on quasispecies , the genetic distance within one cluster may help to determine the infection time and immune escaping.The analysis of related affecting factors need more samples.
