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目的使用两种HIV-1基因型耐药检测方法分别检测干血斑(Dried Blood spot,DBS)样本并比较耐药结果,以期建立适合的检测方法。方法选择河南省和安徽省经高效抗逆转录病毒治疗(highly active anti-retroviral therapy,HAART)5年以上HIV感染者共35名,采集全血制作干血斑标本,进行CD4+细胞计数和病毒载量测定,分别使用两种文献报道的方法扩增干血斑标本中的HIV-1 pol区核苷酸片段,进行HIV-1基因型耐药检测,并比较34个主要耐药突变位点差异。结果低载量<1×103copies/m L,方法2具有更高的扩增率(5/5),而方法1能扩增成功3份(3/5)。方法2共检出269个主要耐药突变位点,两种方法耐药位点的一致率为95.9%(258/269)。与方法1比较,有11个耐药位点存在差异,7个位点由混合碱基引起,另4个差异位点则集中为:E138Q和K101Q。结论两种方法都能够检测病毒载量大于1×103copies/m L的样本,但是对于低载量样本方法 2较方法 1有更好的表现,后续应选择更多的不同亚型标本进行研究。
Objective To detect two samples of DRS (resistant blood spot, DBS) and compare drug resistance with two HIV-1 genotypic drug resistance test methods in order to establish a suitable detection method. Methods Thirty-five HIV-positive patients with highly active anti-retroviral therapy (HAART) in Henan and Anhui Provinces were collected. Whole blood was collected to make dry blood spot samples for CD4 + cell count and viral load Two kinds of reported methods were used to amplify nucleotide fragments of HIV-1 pol in the dried blood spot samples, and the HIV-1 genotypes were tested for drug resistance. The difference of 34 major drug-resistant mutation sites was compared . Results Low loading <1 × 103 copies / m L, method 2 had a higher rate of amplification (5/5), and method 1 successfully amplified 3 (3/5). Twenty-two out of 269 major resistance mutations were detected in method 2, and the agreement between the two methods was 95.9% (258/269). Compared with the method 1, there are 11 resistance sites there are differences, seven sites caused by the mixed base, the other four differences are concentrated at: E138Q and K101Q. Conclusions Both methods are capable of detecting samples with a viral load greater than 1 × 10 3 copies / m L but perform better with method 1 for low-load samples and more subtypes later.