异源双链泳动分析法在人类免疫缺陷病毒1基因亚型分析中的应用  

Application of heteroduplex mobility assay in genetic subtyping on human immunodeficiency virus type 1

作　　者：陈亮[1] 严延生[1] 翁育伟[1] 王惠榕[1] 吴守丽[1] 陈舸[1] 郑兆双[1] 郑健[1] 颜苹苹[1] 

机构地区：[1]福建省卫生防疫站艾滋病防治研究室,福州350001

出　　处：《中华流行病学杂志》2002年第2期127-130,共4页Chinese Journal of Epidemiology

摘　　要：目的　运用异源双链泳动分析法 (HMA )进行人类免疫缺陷病毒 1(humanimmunodeficiencyvirus 1,HIV 1)基因亚型分析 ,了解福建省HIV 1亚型的流行情况。方法　收集细胞培养中HIV 1感染的外周血单个核细胞 (PBMC) ,聚合酶链反应 (PCR)扩增HIV 1膜蛋白基因(env)区的核酸片段 ,与标准亚型的对照质粒的PCR产物进行杂交 ,形成异源双链二聚体 ,根据其在聚丙烯酰胺凝胶电泳中的泳动率确定其亚型 ,并进一步对标本进行序列测定和系统树分析。结果15份标本中 ,80 .0 0 %为E亚型 ,6 .6 7%为B亚型 ,2份不能确定其亚型。比较异源双链泳动法和序列测定法的结果 ,显示两种分析方法对HIV 1病毒基因分型有较高的一致性 ,其亚型一致率为86 .6 7%。结论　福建省流行的HIV 1毒株以E亚型为主。亚型分析法具有快速、简便、经济和高特异性的特点 ,可用作对HIV流行毒株进行长期监测的重要手段。Objective Using heteroduplex mobility assay(HMA) to subtype human immunodeficiency virus type 1(HIV 1) for the purpose of understanding HIV 1 subtype epidemic in Fujian province. Methods DNA fragments of HIV 1 env gene were amplified from peripheral blood mononuclear cell(PBMC) cocultures of HIV 1 infected individuals by nested polymerase chain reation(PCR). Heteroduplexs were formed through hybridizing PCR products from the samples and reference plasmid.According to the mobility of heteroduplexs in polyacrylamide gel electrophoresis, HIV 1 subtype from that sample was characterized and further confirmed by nucleotide sequencing analysis. Results Thirteen of 15( 86.67 %) samples were successfully subtyped by HMA,except 2 failures. Subtype E and B took up 80%(12/15) and 6.67 %(1/15) respectively. Results indicated a high concordance between HMA and nucleotide sequencing analysis and concordance rate was 86.67 %(13/15). Conclusions Subtype E appeared to be the major epidemic strain of HIV 1 in Fujian. HMA showed the characteristics of fastness, easiness, economie and with high specificity, and can be used in the survillence for the epidemic strain of HIV 1.

关 键 词：异源双链泳动分析法 人类免疫缺陷病毒1 基因亚型 HIV-1 序列测定 

分 类 号：R373[医药卫生—病原生物学]