A multiplex liquid-chip assay based on Luminex xMAP technology for simultaneous detection of six common respiratory viruses

// Yong Yan 1, * , Jian-Yong Luo 1, * , Yin Chen 2, * , Heng-Hui Wang 1 , Guo-Ying Zhu 1 , Pei-Yan He 1 , Jin-Lei Guo 1 , Yong-Liang Lei 3 and Zhong-Wen Chen 1 1 Jiaxing Key Laboratory of Pathogenic Microbiology, Jiaxing Municipal Center for Disease Control and Prevention, Jiaxing 314050, China 2 Institute of Microbiology, Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou 310051, China 3 Lishui Municipal Center for Disease Control and Prevention, Lishui 323000, China * These authors contributed equally to this work Correspondence to: Zhong-Wen Chen, email: heton74@hotmail.com Keywords: Luminex, xMAP, respiratory virus, real-time RT-PCR, multiplex detection Received: December 30, 2016      Accepted: June 02, 2017      Published: June 17, 2017 ABSTRACT 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.