Annals of Clinical Microbiology, The official Journal of the Korean Society of Clinical Microbiology

Comparison Cytomegalovirus Qualitative Assay Using Real-Time PCR and Conventional PCR

Original article

Annals of Clinical Microbiology (Ann Clin Microbiol) 2013 March, Volume 16, Issue 1, pages 19-24.

https://doi.org/10.5145/ACM.2013.16.1.19

Comparison Cytomegalovirus Qualitative Assay Using Real-Time PCR and Conventional PCR

Seri Jeong1, Yoon Jung Kim1, Il Kwon Bae1, Moon Jung Kim2, Seok Hoon Jeong1
1Department of Laboratory Medicine and Research Institute of Bacterial Resistance, Yonsei University College of Medicine, Seoul, 2Department of Biomedical Sciences, Graduate School of Public Health Science, Eulji University, Daejeon, Korea

Abstract

Background: Cytomegalovirus (CMV) infection is a major cause of morbidity and mortality in immunocompromised patients. We compared the abilities of the recently developed Real-Q Cytomegalovirus Kit (Biosewoom Inc., Korea) and the previously used PANA mPCR CMV Detection Kit (Panagene Inc., Korea) to detect CMV.

Methods: We analyzed 300 samples (whole blood: 262, urine: 37, CSF: 1) submitted for qualitative CMV PCR testing during October 2011 at Yonsei University College of Medicine Severance Hospital. real-time PCR was performed with a Real-Q Cytomegalovirus Kit and conventional PCR was conducted with a PANAmPCR CMV Detection Kit.

Results: The positive rates of both real-time PCR and conventional PCR were 25.3% (76/300), and the kappa coefficient (K) was 0.96 (95% confidence interval (CI), 0.93-1.00). The concordance rate of the Real-Q Cytomegalovirus Kit and the PANAmPCR CMV Detection Kit was 98.7% (296/300), and four out of 300 samples showed discordant results. If the concordant results of 296 samples and the four results confirmed by direct sequencing were assumed to be true, the sensitivity and specificity of the Real-Q Cytomegalovirus Kit were 97.4% (95% CI, 93.8-100.0%) and 99.1% (95% CI, 97.9-100.0%), respectively.

Conclusion: The recently developed Real-Q Cytomegalovirus Kit showed excellent sensitivity and specificity, and had a high concordance rate with the previously established PANAmPCR CMV Detection Kit, which uses conventional PCR. Furthermore, real-time PCR could decrease the test time, as the electrophoresis step required for conventional PCR is not required for real-time PCR. (Ann Clin Microbiol 2013; 16:19-24)

Keywords

Cytomegalovirus (CMV), Polymerase chain reaction (PCR), Real-time polymerase chain reaction