INNO-LiPA和直接测序法检测HBV耐药突变的比较

Comparison of sequence analysis and the INNO-LiPA HBV DR line probe assay for detection of resistant HBV strains

目的比较INNO-LiPA HBV DRv2和直接测序法检测HBV耐药突变的准确性和灵敏性。方法选取41例接受阿德福韦或聚乙二醇干扰素α-2a救援治疗并在治疗过程中(治疗48周和72周时)进行INNO-LiPA HBV DRv2耐药检测的拉米夫定耐药患者,共收集到101份足量的血清,采用直接测序法进行拉米夫定和阿德福韦耐药检测。分析HBV聚合酶第80、173、180、181、204和236位点的耐药突变情况,并与INNO-LiPA耐药检测的结果进行对比。结果 INNO-LiPA HBV DRv2和直接测序法检测204位点耐药结果对比,完全符合率为73.8%(62/84),部分符合率为23.8%(20/84),总体符合率达到97.6%(82/84)。其他位点总体符合率也多在95%以上。另外INNO-LiPA比直接测序法有更高的灵敏度,对混合感染的检出率(46.4%)明显高于直接测序法(23.8%),其总体耐药检出率也高于直接测序法。结论 INNO-LiPA方法在灵敏度和准确性上均优于直接测序法,而且操作简便、快捷,能够提供更多的信息量,在科研和临床上都有很好的应用前景。

Objective＇ To assess the accuracy and sensitivity of INNO-LiPA HBV DR v2 for detecting drug resistance strains of hepatitis B virus. Methods One hundred and one sera samples from 41 lamivudine-resistant chronic hepatitis B patients undergoing rescue therapy were collected. Drug resistant mutations of the reverse transcriptase region of the HBV Polymerase gene had been determined using INNO-LiPA at baseline, week 48 and week 72 during the therapy. For further study, the resistant mutations were detected using direct sequencing. The results obtained for codons 80, 173, 180, 181, 204, and 236 were compared between sequence analysis and INNO-LiPA. Results Identical results were found with 82 of the 84 samples （97.6%） between the results of LiPA and sequence analysis, with 62 serum samples were fully concordant （73.8%） and 20 serum samples were partially concordant （23.8%）. It showed higher sensitivity than direct sequencing, particularly for the detection of mixed drug-resistance mutations. Conchmion The INNO-LiPA HBV DR v2 appears to be a more sensitive and rapid assay for detection of HBV genotypic resistance.