中文摘要：

背景临床的管理精确地并且易感知地为次要的 HIV-1 抵抗变化测试是很重要的。HIV 药抵抗察觉的常规 genotypic 试金能仅仅基于定序区别在超过 20%-30% 介绍的变化。学习是评估等位基因特定的即时 PCR ( ASPCR )检测抵抗相关的变化的这的目的在位置定位了我们开发了的 103 ， 184 和 215 .Methods 等位基因特定的 PCR 试金用在中国爱滋病病人的最普通的药抵抗变化， K103N ， M184V/I ， T215F/Y 作为一个模型系统。标准被克隆野类型、变异的 DNA 碎片进 T 向量构造。我们设计了特定的教材作为一个荧光记者用 SYBR 绿色在即时 PCR 区别变异的模板。然后我们评估了 ASPCR 试金并且用 ASPCR 试金的敏感是的这 method.Results 测试了 140clinical 样品 0.04% 为 K103N， 0.30% 为 M1841， 0.40% 为 M184V， 0.03% 为 T215F 并且 0.02% 为 T215Y。intra 试金和变化的内部试金的系数是不到 0.42。140 件血浆样品被 ASPCR 测试，十个病人的动态抵抗曲线是 obtained.Conclusions 药抵抗出现在 antiretroviral 的开始以后的半年治疗。到在开始治疗以后的 1.5 年的 T215Yemerged 1 的变化然后很快增加了。我们开发了的 ASPCR 试金是一个敏感、精确、快速的方法检测次要的 HIV-1 变体，它能更早并且更为 HIV 研究和爱滋病 antiretroviral 治疗提供药抵抗信息。

英文摘要：

Background It is very important for the clinical management to test for minor HIV-1 resistance mutations accurately and sensitively. The conventional genotypic assays of HIV drug resistance detection based on sequencing can only discriminate the mutations which present in more than 20%-30%. The aim of this study was to evaluate allele-specific real-time PCR （ASPCR） to detect the resistance-related mutations located at positions 103, 184 and 215.Methods We developed the allele-specific PCR assay, using the most common drug resistance mutations in Chinese AIDS patients, K103N, M184V/I, T215F/Y as a model system. The standards were constructed by cloning the wild-type and mutant DNA fragments into the T-vector. We designed specific primers to discriminate mutant templates in the real-time PCR using SYBR green as a fluorescence reporter. And then we evaluated the ASPCR assay and tested 140clinical samples using this method.Results The sensitivities of ASPCR assay were 0.04% for K103N, 0.30% for M1841, 0.40% for M184V, 0.03% for T215F and 0.02% for T215Y. The intra-assay and inter-assay coefficients of variation were less than 0.42. One hundred and forty plasma samples were tested by ASPCR and dynamic resistance curves of ten patients were obtained.Conclusions Drug resistance emerged half a year after the start of antiretroviral therapy. The mutation of T215Yemerged 1 to 1.5 years after starting treatment and then increased rapidly. The ASPCR assay we developed was a sensitive, accurate and rapid method to detect the minor HIV-1 variants and it can provide earlier and more drug-resistance information for HIV research and AIDS antiretroviral therapy.