新型HIV-1表型耐药系统临床检测应用研究

Clinical detection of HIV-1 drug resistance using a novel HIV-1 phenotypic resistance system

目的应用新型HIV-1表型耐药系统对经过HAART治疗的40例临床患者样本进行表型耐药检测,通过比较表型和基因型耐药的结果,实现这一新型HIV-1表型耐药体系在临床检测中的应用。方法通过分子克隆方法、Gateway重组技术、体外细胞功能试验,得到表型耐药结果。结果成功构建含有患者基因的用于假病毒表型耐药的假病毒株40株,在总数达54个表型耐药性检测中,与基因型耐药性检测一致性达75.9%(41/54)。其中蛋白酶抑制剂(克力芝)试验组、核苷类逆转录酶抑制剂(3TC)试验组和非核苷类逆转录酶抑制剂(EFV)试验组的表型耐药性检测,与基因型耐药性检测一致性高达100.0%;而核苷类逆转录酶抑制剂(AZT)试验组共6个表型耐药性检测,与基因型耐药性检测一致性为66.7%(4/6);2组多药试验共24个表型耐药性检测,与基因型耐药性检测一致性为54.2%(13/24)。其中显示出一个较为明显的特征,AZT药物敏感性试验的表型检测结果大多低于其基因型耐药检测的耐药水平(68.75%,11/16)。结论本研究建立了一个新型表型耐药系统对HIV耐药性进行检测,通过40例临床样本的检测,充分证明新型HIV-1表型耐药检测体系可以准确地反应样本耐药水平并对其进行定量。这一方法弥补了以往假病毒表型耐药检测方法中载体不能重组所有药物靶点的缺陷,证明该新型HIV-1表型耐药体系具有广阔的临床检测应用前景。

Objective To evaluate a novel phenotypic assay in the clinical detection of HIV-1drug resistance.Methods By using molecular cloning and gateway recombinant techniques to identify HIV-1drug inhibition and phenotypic resistance.Results A total of 40 pseudovirus strains were constructed for the phenotypic assay and 54 clinical samples were detected.The concordance between phenotypic and genotypic assays reached75.9%（41/54）.In the groups of PRIs,3TC and EFV resistance assays,the concordance between phenotype and genotype was 100.0%,66.7%（4/6）in the group of AZT assay,and 54.2%（13/24）in the group of multi-drug assay.The results showed that the phenotypic susceptibility to AZT were significantly lower than genotypic susceptibility 68.75%,11/16）.Conclusions A novel phenotype assay for detecting HIV drug resistance was established.By testing 40 clinical samples,it was proved that this phenotypic system can detect drug resistance at high accuracy.Phenotype assay could be the supplementary for predicting HIV drug resistance in clinical evaluation.