Yan Yong, Luo Jian-Yong, Chen Yin, Wang Heng-Hui, Zhu Guo-Ying, He Pei-Yan, Guo Jin-Lei, Lei Yong-Liang, Chen Zhong-Wen
Jiaxing Key Laboratory of Pathogenic Microbiology, Jiaxing Municipal Center for Disease Control and Prevention, Jiaxing 314050, China.
Institute of Microbiology, Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou 310051, China.
Oncotarget. 2017 Jun 17;8(57):96913-96923. doi: 10.18632/oncotarget.18533. eCollection 2017 Nov 14.
We utilized one-step multiplex reverse transcription-PCR (RT-PCR) and Luminex xMAP technology to develop a respiratory multiplex liquid-chip assay (rMLA) for simultaneous detection of 6 common respiratory viruses, including influenza virus type A (FluA) and type B (FluB), para-influenza virus type 3 (PIV-3), respiratory syncytial virus (RSV), human metapneumovirus (MPV) and a threatening virus to China, Middle East Respiratory Syndrome coronavirus (MERS-CoV). Performance of rMLA was evaluated by comparing with real-time RT-PCR. Detection data from clinical specimens showed that the rMLA had diagnostic sensitivities of 97.10% for FluA, 94.59% for FluB, 98.68% for PIV-3, 94.87% for RSV and 95.92% for MPV (No Data for MERS-CoV due to the lack of positive specimens). Data of analytical sensitivities showed that the detection limits of the rMLA assay were 5-25 viral RNA copies per μl for FluA, FluB, PIV-3 and MERS-CoV, approximate to the real-time RT-PCR assay; while the values were 8 and 22copies/μl for MPV and RSV, lower than the real-time RT-PCR(78 and 114 copies/μl respectively). The results indicated that the rMLA is a sensitive, specific detection tool and comparable to real-time RT-PCR, especially suitable for high-throughput detection of respiratory specimens.
我们利用一步多重逆转录聚合酶链反应（RT-PCR）和Luminex xMAP技术开发了一种呼吸道多重液相芯片检测方法（rMLA），用于同时检测6种常见呼吸道病毒，包括甲型流感病毒（FluA）、乙型流感病毒（FluB）、3型副流感病毒（PIV-3）、呼吸道合胞病毒（RSV）、人偏肺病毒（MPV）以及对中国构成威胁的病毒——中东呼吸综合征冠状病毒（MERS-CoV）。通过与实时RT-PCR进行比较，评估了rMLA的性能。临床标本的检测数据显示，rMLA对FluA的诊断敏感性为97.10%，对FluB为94.59%，对PIV-3为98.68%，对RSV为94.87%，对MPV为95.92%（由于缺乏阳性标本，MERS-CoV无数据）。分析敏感性数据显示，rMLA检测方法对FluA、FluB、PIV-3和MERS-CoV的检测限为每微升5 - 25个病毒RNA拷贝，与实时RT-PCR检测方法相近；而MPV和RSV的值分别为8和22拷贝/微升，低于实时RT-PCR（分别为78和114拷贝/微升）。结果表明，rMLA是一种灵敏、特异的检测工具，与实时RT-PCR相当，特别适合呼吸道标本的高通量检测。
