新型国产乙型肝炎病毒核酸定量检测试剂的质量评价

Evaluation of a novel domestic hepatitis B viral DNA quantitative fluorescence diagnostic kit

目的评价新型国产乙型肝炎病毒核酸定量检测试剂（以下简称“新型定量试剂”）的质量。方法首先用基于磁珠法自动化核酸提取平台的新型定量试剂和Roche定量试剂对WHO的HBVDNA系列稀释标准品（50、200、2000、20000 IU／ml）进行溯源性分析，确定定量试剂检测的可靠性。然后，用2种定量试剂对HBVB和C基因型标准血浆（分别稀释为25、50、200、2000、20000、200000 IU／ml）分别在3个不同检测中心共进行216次平行检测，比较其准确度和精密度。最后，用2种定量试剂平行检测5份HBV DNA阴性血浆样本和37份不同病毒载量的HBV DNA阳性临床血浆标本，并评价其相关性和符合率。结果经溯源性分析，2种定量试剂在上述4个检测节点的实测对数值与理论对数值的差值偏差均在可接受范围之内（0．005～0．280 Ig IU／ml）。2种定量试剂测定各检测节点变异系数的结果经配对比较，HBV的B、C基因型血浆检测的精密度差异均无统计学意义（B基因型：t=1．226，P=0．275；C基因型，t=2．319，P=0．07）。2种定量试剂检测42份临床血浆标本的总符合率达97，62％（41／42），且检测37份HBV DNA阳性血浆标本结果显著相关（R^2=0．934，P〈0．0001）。结论新型定量试剂与Roche定量试剂在溯源性、符合率、精密度和血清盘评价中的差异无显著性，该定量试剂有良好的检测质量，可用于临床HBVDNA检测。（中华检验医学杂志，2013，36：280-285）

Objective Comparative performance characteristics of a novel domestic （Hunan Sansure） HBV DNA quantitative fluorescence diagnostic kit （PCR-fluorescence probing） and its counterpart Roche Cobas Ampliprep/COBAS Taqman HBV test kit. Methods Firstly, WHO international standard-for HBV-DNA was diluted to 50, 200, 2000, 20 000 IU/ml separately, and then all diluted samples were detected by both the novel domestic HBV DNA quantitative fluorescence diagnostic kit and Roche Cobas Ampliprep/COBAS Taqman HBV test kit to perform traceability analysis and to determine kits＇reliability. Secondly, pre-diluted series of standard HBV B and C genotype plasma （25, 50, 200, 2000, 20 000,200 000 IU/ml） were simultaneously performed 216 times for three-center detection by use of the domestic and Roche HBV DNA test kits separately. Accuracy and precision of those two types of HBV DNA kits were comparatively analyzed. Finally, a total of 42 clinical plasma samples including 5 negative HBV DNA and different concentration levels of 37 positive HBV DNA were detected by the above two types of kits to perform clinical quality evaluation. Results Traceability results showed that both HBV DNA test kits agreed with the HBV DNA WHO standard across all four titers tested and all absolute differences （observed mean HBV HBV-expected HBV DNA ） were within O. 3 lg IU/ml. Accuracy results indicated that the deviation of all observed values at 6 titers （ both of HBV genotyping B and C ） tested were within the acceptance criteria （0. 005 -0. 280 lg IU/ml）. Comparative performance of coefficient of variation of PCR assays to HBV genotypes B and C measured by both kits showed that there were no differences of precision were found （ genotyping B： t = 1. 226, P = O. 275; genotyping C： t = 2. 319, P = O. 07 ） . The clinical performance of the domestic assay compared to the COBAS assay had been assessed on a panel of 42 clinical specimens. The qualitative results indicated that the total concordant results between two tests were determined in 97.62% （41/42） of samples. Also, a significant correlation was observed between values tested by two HBV DNA kits in 37 HBV DNA positive samples （ R^2=0. 934, P 〈 0. 0001 ）. Conclusion No significant differences of the traceability, accuracy, precision and clinical detection are found between two kits. （Chin J Lab Med, 2013,36：280-285）