Evaluation of the Efficacies of Rapid Antigen Test, Multiplex PCR, and Real-time PCR for the Detection of a Novel Influenza A (H1N1) Virus

Yusun Hwang; Kyounghee Kim; Miae Lee

doi:10.3343/kjlm.2010.30.2.147

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Hwang, Kim, and Lee: Evaluation of the Efficacies of Rapid Antigen Test, Multiplex PCR, and Real-time PCR for the Detection of a Novel Influenza A (H1N1) Virus

Original Article

Korean J Leg Med 2010;30(2):147-152.

Published online: 14 January 2010

DOI: https://doi.org/10.3343/kjlm.2010.30.2.147

Evaluation of the Efficacies of Rapid Antigen Test, Multiplex PCR, and Real-time PCR for the Detection of a Novel Influenza A (H1N1) Virus

Yusun Hwang, M.D., Kyounghee Kim, M.T., Miae Lee, M.D.

Department of Laboratory Medicine, School of Medicine, Ewha Womans University, Seoul, Korea

Corresponding author: Miae Lee, M.D. Department of Laboratory Medicine, Mokdong Hospital, School of Medicine, Ewha Womans University, 911-1 Mok-dong, Yangcheon-gu, Seoul 158-710, Korea Tel: +82-2-2650-5222, Fax: +82-2-2650-5222 E-mail: miae@ewha.ac.kr

Revised 19 November 200912 February 2010 Accepted 18 February 2010

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

Background:

In April 2009, a novel influenza A (H1N1) virus was detected in the US, and at the time of conducting this study, H1N1 infection had reached pandemic proportions. In Korea, rapid antigen tests and PCR assays have been developed to detect the H1N1 virus. We evaluated the efficacies of rapid antigen test, multiplex PCR, and real-time PCR for detecting the H1N1 virus.

Methods:

From August to September 2009, we tested 734 samples obtained from nasopharyngeal swab or nasal swab using rapid antigen test (SD Influenza Antigen, Standard Diagnostics, Inc., Korea) and multiplex PCR (Seeplex FluA ACE Subtyping, Seegene, Korea). We also tested 224 samples using the AdvanSure real-time PCR (LG Life Sciences, Korea) to compare the results obtained using real-time PCR with those obtained using multiplex PCR. Furthermore, 99 samples were tested using the AdvanSure real-time PCR and the AccuPower real-time PCR (Bioneer, Korea).

Results:

In comparison with the results of multiplex PCR, the sensitivity and specificity of the rapid antigen test were 48.0% and 99.8%, respectively. The concordance rate for multiplex PCR and the AdvanSure real-time PCR was 99.6% (κ=0.991, P=0.000), and that for the AdvanSure real-time PCR and the AccuPower real-time PCR was 97.0% (κ=0.936, P=0.000).

Conclusions:

The rapid antigen test is significantly less sensitive than PCR assay; therefore, it is not useful for H1N1 detection; however multiplex PCR, the AdvanSure real-time PCR, and the AccuPower real-time PCR can be useful for H1N1 detection.

Keywords: Novel influenza A (H1N1) virus, Rapid antigen test, Multiplex PCR, Real-time PCR

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Table 1.

Comparison of the efficacies of the rapid antigen test and multiplex PCR for detecting the novel influenza A (H1N1) virus

		Multiplex PCR		Total
		+	-	Total
Rapid antigen test	+	49	1∗	50
	-	53	631^†	684
Total		102	632	734

^∗ One case showing a positive result in rapid antigen test was negative for novel influenza A (H1N1), but positive for seasonal influenza A (H3) in multiplex PCR;

^† including 5 cases of seasonal influenza A (H3) and 1 case of seasonal influenza A (H1) detected using multiplex PCR.

Table 2.

Comparison of the efficacies of multiplex PCR and AdvanSure real-time PCR for detecting the novel influenza A (H1N1) virus

		Multiplex PCR		Total
		+	-	Total
AdvanSure real-time PCR	+	105	1∗	106
	-	0	118	118
Total		105	119	224

^∗ Novel influenza A (H1N1) was confirmed by sequencing.

Table 3.

Comparison of the efficacies of AdvanSure real-time PCR and AccuPower real-time PCR for detecting the novel influenza A (H1N1) virus

		AdvanSure real-time PCR		Total
		+	-	Total
AccuPower real-time PCR	+	59	2∗	61
	-	1∗	37	38
Total		60	39	99

^∗ Sequencing revealed 3 cases of novel influenza A (H1N1).

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