Sun Yu, Jia Tingting, Sun Yanli, Han Yanxi, Wang Lunan, Zhang Rui, Zhang Kuo, Lin Guigao, Xie Jiehong, Li Jinming
National Center for Clinical Laboratory, Beijing Hospital of National Health and Family Planning Commission, Beijing, People's Republic of China.
J Clin Microbiol. 2013 Dec;51(12):4055-9. doi: 10.1128/JCM.02018-13. Epub 2013 Oct 2.
An external quality assessment (EQA) program for the molecular detection of avian influenza A (H7N9) virus was implemented by the National Center for Clinical Laboratories (NCCL) of China in June 2013. Virus-like particles (VLPs) that contained full-length RNA sequences of the hemagglutinin (HA), neuraminidase (NA), matrix protein (MP), and nucleoprotein (NP) genes from the H7N9 virus (armored RNAs) were constructed. The EQA panel, comprising 6 samples with different concentrations of armored RNAs positive for H7N9 viruses and four H7N9-negative samples (including one sample positive for only the MP gene of the H7N9 virus), was distributed to 79 laboratories in China that carry out the molecular detection of H7N9 viruses. The overall performances of the data sets were classified according to the results for the H7 and N9 genes. Consequently, we received 80 data sets (one participating group provided two sets of results) which were generated using commercial (n = 60) or in-house (n = 17) reverse transcription-quantitative PCR (qRT-PCR) kits and a commercial assay that employed isothermal amplification method (n = 3). The results revealed that the majority (82.5%) of the data sets correctly identified the H7N9 virus, while 17.5% of the data sets needed improvements in their diagnostic capabilities. These "improvable" data sets were derived mostly from false-negative results for the N9 gene at relatively low concentrations. The false-negative rate was 5.6%, and the false-positive rate was 0.6%. In addition, we observed varied diagnostic capabilities between the different commercially available kits and the in-house-developed assays, with the assay manufactured by BioPerfectus Technologies (Jiangsu, China) performing better than the others. Overall, the majority of laboratories have reliable diagnostic capacities for the detection of H7N9 virus.
中国国家临床检验中心（NCCL）于2013年6月实施了一项针对甲型禽流感（H7N9）病毒分子检测的外部质量评估（EQA）计划。构建了包含H7N9病毒血凝素（HA）、神经氨酸酶（NA）、基质蛋白（MP）和核蛋白（NP）基因全长RNA序列的病毒样颗粒（VLP）（铠装RNA）。该EQA检测板包含6个不同浓度的H7N9病毒铠装RNA阳性样本和4个H7N9阴性样本（包括1个仅H7N9病毒MP基因阳性的样本），分发给中国79个开展H7N9病毒分子检测的实验室。根据H7和N9基因的检测结果对数据集的整体表现进行分类。因此，我们收到了80个数据集（一个参与组提供了两组结果），这些数据集是使用商业（n = 60）或自制（n = 17）逆转录定量PCR（qRT-PCR）试剂盒以及一种采用等温扩增方法的商业检测方法（n = 3）生成的。结果显示，大多数（82.5%）数据集正确鉴定出了H7N9病毒，而17.5%的数据集在诊断能力方面需要改进。这些“可改进”的数据集大多来自相对低浓度下N9基因的假阴性结果。假阴性率为5.6%，假阳性率为0.6%。此外，我们观察到不同市售试剂盒和自制检测方法之间的诊断能力存在差异，由江苏百盛医疗科技有限公司生产的检测方法表现优于其他方法。总体而言，大多数实验室在检测H7N9病毒方面具有可靠的诊断能力。
