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Ann Lab Med. Nov 2012; 32(6): 399–406.
Published online Oct 17, 2012. doi:  10.3343/alm.2012.32.6.399
PMCID: PMC3486933
Evaluation of a Multiplex Real-time PCR Assay for the Detection of Respiratory Viruses in Clinical Specimens
Insoo Rheem, M.D.,1 Joowon Park, M.D.,1 Tae-Hyun Kim, Ph.D.,2 and Jong Wan Kim, M.D.corresponding author1
1Department of Laboratory Medicine, Dankook University College of Medicine, Cheonan, Korea.
2Institute of Tissue Regeneration Engineering (ITREN), Dankook University, Cheonan, Korea.
corresponding authorCorresponding author.
Corresponding author: Jong Wan Kim. Department of Laboratory Medicine, Dankook University College of Medicine, 119 Dandae-ro, Dongnam-gu, Cheonan 330-715, Korea. Tel: +82-41-550-6662, Fax: +82-41-555-7155, wan1818/at/
Received January 12, 2012; Revised April 30, 2012; Accepted August 17, 2012.
In this study, we evaluated the analytical performance and clinical potential of a one-step multiplex real-time PCR assay for the simultaneous detection of 14 types of respiratory viruses using the AdvanSure RV real-time PCR Kit (LG Life Sciences, Korea).
Three hundred and twenty clinical specimens were tested with the AdvanSure RV real-time PCR Kit and conventional multiplex reverse transcription (RT)-PCR assay. The assay results were analyzed and the one-step AdvanSure RV real-time PCR Kit was compared with the conventional multiplex RT-PCR assay with respect to the sensitivity and specificity of the detection of respiratory viruses.
The limit of detection (LOD) was 1.31 plaque-forming units (PFU)/mL for human rhinoviruses (hRVs), 4.93 PFU/mL for human coronavirus HCoV-229E/NL63, 2.67 PFU/mL for human coronavirus HCoV-OC43, 18.20 PFU/mL for parainfluenza virus 1 (PIV)-1, 24.57 PFU/mL for PIV-2, 1.73 PFU/mL for PIV-3, 1.79 PFU/mL for influenza virus group (Flu) A, 59.51 PFU/mL for FluB, 5.46 PFU/mL for human respiratory syncytial virus (hRSV)-A, 17.23 PFU/mL for hRSV-B, 9.99 PFU/mL for human adenovirus (ADVs). The cross-reactivity test for this assay against 23 types of non-respiratory viruses showed negative results for all viruses tested. The agreement between the one-step AdvanSure multiplex real-time PCR assay and the conventional multiplex RT-PCR assay was 98%.
The one-step AdvanSure RV multiplex real-time PCR assay is a simple assay with high potential for specific, rapid and sensitive laboratory diagnosis of respiratory viruses compared to conventional multiplex RT-PCR.
Keywords: Respiratory virus, Multiplex real-time PCR, Limit of detection
Articles from Annals of Laboratory Medicine are provided here courtesy of
Korean Society for Laboratory Medicine