A multiplex liquid-chip assay based on Luminex xMAP technology for simultaneous detection of six common respiratory viruses
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Yong Yan1,*, Jian-Yong Luo1,*, Yin Chen2,*, Heng-Hui Wang1, Guo-Ying Zhu1, Pei-Yan He1, Jin-Lei Guo1, Yong-Liang Lei3 and Zhong-Wen Chen1
1Jiaxing Key Laboratory of Pathogenic Microbiology, Jiaxing Municipal Center for Disease Control and Prevention, Jiaxing 314050, China
2Institute of Microbiology, Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou 310051, China
3Lishui Municipal Center for Disease Control and Prevention, Lishui 323000, China
*These authors contributed equally to this work
Zhong-Wen Chen, email: [email protected]
Keywords: Luminex, xMAP, respiratory virus, real-time RT-PCR, multiplex detection
Received: December 30, 2016 Accepted: June 02, 2017 Published: June 17, 2017
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.
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