Ann Lab Med 2018; 38(1): 46-50  
Evaluation of Allplex Respiratory Panel 1/2/3 Multiplex Real-Time PCR Assays for the Detection of Respiratory Viruses with Influenza A Virus subtyping
Jaehyeon Lee, M.D.1,3, Hye Soo Lee, M.D.2,3, Yong Gon Cho, M.D.2,3, Sam Im Choi, M.D.2,3, and Dal Sik Kim, M.D.2,3
Department of Laboratory Medicine1, Chonbuk National University Hospital, Jeonju; Department of Laboratory Medicine2, Chonbuk National University Medical School, Jeonju; Research Institute of Clinical Medicine of Chonbuk National University-Biomedical Research Institute of Chonbuk National University Hospital3, Jeonju, Korea
Correspondence to: Dal Sik Kim
Department of Laboratory Medicine, Chonbuk National University Medical School, 20 Geonji-ro, Dukjin-gu, Jeonju 54907, Korea
Tel: +82-63-250-1793
Fax: +82-63-250-1200
E-mail: dskim@jbnu.ac.kr
Received: March 20, 2017; Revised: April 27, 2017; Accepted: September 16, 2017; Published online: January 1, 2018.
© The Korean Society for Laboratory Medicine. All rights reserved.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
The Allplex Respiratory Panel 1/2/3 (All16) is a multiplex PCR assay for detecting 16 respiratory viruses with influenza A virus (FluA) subtyping, and the first clinical assay based on multiple detection temperatures. We compared the results between All16 and Anyplex II RV16 (Any16) in 426 clinical samples. Samples showing discrepancies between the two tests were further tested using monoplex PCR. FluA subtyping based on the hemagglutinin type results of All16, which yielded H1, H3, and non-H1/H3, was compared with the results of the BioFire FilmArray respiratory panel. The positive and negative percent agreements and kappa value for each virus between All16 and Any16 ranged from 54.5-100.0%, 84.7-100.0%, and 0.57-1.00, respectively. FluA subtype results from All16 for 26 samples were consistent with those from FilmArray. Good agreement was observed between the two methods, except when analyzing human enterovirus (kappa value 0.70), and the All16 showed reliable FluA subtyping results. For parainfluenza virus 3, the All16 was more sensitive than Any16. When testing 28 samples simultaneously, the mean test time and hands-on time were 4.3 and 0.5 hours, respectively in All16. In conclusion, All16 showed reliable performance, but further studies are needed regarding human enterovirus analysis.
Keywords: Respiratory Virus, Multiplex real-time PCR, Reverse transcriptase-PCR, Influenza A subtyping, Evaluation


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