Ann Lab Med.  2018 Jan;38(1):46-50. 10.3343/alm.2018.38.1.46.

Evaluation of Allplex Respiratory Panel 1/2/3 Multiplex Real-Time PCR Assays for the Detection of Respiratory Viruses with Influenza A Virus subtyping

Affiliations
  • 1Department of Laboratory Medicine, Chonbuk National University Hospital, Jeonju, Korea.
  • 2Department of Laboratory Medicine, Chonbuk National University Medical School, Jeonju, Korea. dskim@jbnu.ac.kr
  • 3Research Institute of Clinical Medicine of Chonbuk National University-Biomedical Research Institute of Chonbuk National University Hospital, Jeonju, Korea.

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.

Keyword

Respiratory Virus; Multiplex real-time PCR; Reverse transcriptase-PCR; Influenza A subtyping; Evaluation

MeSH Terms

Enterovirus
Hemagglutinins
Humans
Influenza A virus*
Influenza, Human*
Multiplex Polymerase Chain Reaction
Paramyxoviridae Infections
Polymerase Chain Reaction
Real-Time Polymerase Chain Reaction*
Hemagglutinins

Reference

1. Ginocchio CC. Detection of respiratory viruses using non-molecular based methods. J Clin Virol. 2007; 40(Suppl 1):S11–S14. PMID: 18162248.
2. Mahony JB, Blackhouse G, Babwah J, Smieja M, Buracond S, Chong S, et al. Cost analysis of multiplex PCR testing for diagnosing respiratory virus infections. J Clin Microbiol. 2009; 47:2812–2817. PMID: 19571025.
3. Jorgensen JH, Pfaller MA, editors. American Society for Microbiology. Manual of clinical microbiology. 11th ed. Washington, DC: ASM Press;2015. p. 1432–1435.
4. Lee YJ, Kim D, Lee K, Chun JY. Single-channel multiplexing without melting curve analysis in real-time PCR. Sci Rep. 2014; 4:7439. PMID: 25501038.
5. Fu G, Miles A, Alphey L. Multiplex detection and SNP genotyping in a single fluorescence channel. PLoS One. 2012; 7:e30340. PMID: 22272339.
6. Huang Q, Liu Z, Liao Y, Chen X, Zhang Y, Li Q. Multiplex fluorescence melting curve analysis for mutation detection with dual-labeled, self-quenched probes. PLoS One. 2011; 6:e19206. PMID: 21552536.
7. Huh HJ, Kim JY, Kwon HJ, Yun SA, Lee MK, Lee NY, et al. Performance evaluation of Allplex respiratory panels 1, 2, and 3 for detection of respiratory viruses and influenza A virus subtypes. J Clin Microbiol. 2017; 55:479–484. PMID: 27903601.
8. Huh HJ, Park KS, Kim JY, Kwon HJ, Kim JW, Ki CS, et al. Comparison of the Anyplex II RV16 and Seeplex RV12 ACE assays for the detection of respiratory viruses. Diagn Microbiol Infect Dis. 2014; 79:419–421. PMID: 24985763.
9. Fry AM, Curns AT, Harbour K, Hutwagner L, Holman RC, Anderson LJ. Seasonal trends of human parainfluenza viral infections: United States, 1990-2004. Clin Infect Dis. 2006; 43:1016–1022. PMID: 16983614.
10. Porter DD, Prince GA, Hemming VG, Porter HG. Pathogenesis of human parainfluenza virus 3 infection in two species of cotton rats: Sigmodon hispidus develops bronchiolitis, while Sigmodon fulviventer develops interstitial pneumonia. J Virol. 1991; 65:103–111. PMID: 1845878.
11. Morens DM, Folkers GK, Fauci AS. The challenge of emerging and reemerging infectious diseases. Nature. 2004; 430:242–249. PMID: 15241422.
Full Text Links
  • ALM
Actions
Cited
CITED
export Copy
Close
Share
  • Twitter
  • Facebook
Similar articles
Copyright © 2024 by Korean Association of Medical Journal Editors. All rights reserved.     E-mail: koreamed@kamje.or.kr