检测HIV-1耐药株的异源双链泳动分析技术的建立及临床应用  

Development and clinical application of heteroduplex mobility assay in detecting HIV-1 drug resistance strains

作　　者：姚均[1] 张福杰[1] 赵红心[2] 潘品良[1] 冯鑫[2] 刘志英[2] 蒋岩[1] 

机构地区：[1]中国疾病预防控制中心性病艾滋病预防控制中心,北京100050 [2]北京地坛医院,北京100011

出　　处：《传染病信息》2007年第3期165-167,共3页Infectious Disease Information

基　　金：首都医科发展基金项目(2003-3067)

摘　　要：目的探讨利用异源双链泳动分析技术(HMA)检测HIV-1耐药株的可行性和临床应用。方法从接受高效抗逆转录病毒治疗(HAART)的HIV-1感染者血浆中抽提RNA,采用套式RT-PCR方法扩增HIV-1的PR和RT基因片段,并对扩增的基因片段进行异源双链泳动分析。结果本方法对血清中HIV-1RNA含量2000copies/ml以上的样本都能有效扩增。在18例参加HAART治疗的HIV感染者中,17例的HMA结果在聚丙烯酰氨凝胶电泳上,没有出现有意义的异源双链泳动带。1例发现有异源双链二聚体,这1例后经序列测定证实,确定有耐药位点的基因变异。结论异源双链泳动分析技术能够及时、间接地反映HIV-1在治疗过程中耐药株的存在。该方法的建立能在临床上为临床医生制定治疗方法提供参考依据。Objective To explore the feasibility and clinical application of heteroduplex mobility assay （HMA） in detecting HIV-1 drug resistance strains. Methods HIV-1 viral RNA was extracted from plasma of HIV-infected patients receiving highly active antiretroviral therapy（HAART）. Reverse transcriptase （RT） and protease （PR） genes were amplified with nested RT-PCR, and then analyzed with HMA. Results With HMA, RT and PR genes can be amplified in samples with a minimum 2 000 copies/ml of viral RNA in plasma. Among the 18 HIV-infected patients recieving HAART,17 samples were found no significant heteroduplex bands on the polyacylamid gel. One sample was found heteroduplex band positive and had gene mutation at HIV-1 drug resistant sites following with sequencing. Conclusions HMA can be used to detect the drug resistance strains timely and indirectly during the treament, and the development of this method can offer the reference to the clinical doctors.

关 键 词：HIV-1 耐药株 异源双链泳动分析技术 高效抗逆转录病毒治疗 

分 类 号：R511.1[医药卫生—内科学]