| Title | Simultaneous Detection of Waterborne Viruses by Multiplex Real-Time PCR |
|---|---|
| Author | Lae-Hyung Kang1, Se-hwan Oh1, Jeong-Woong Park2, Yu-Jung Won1, Sangryeol Ryu2, and Soon-Young Paik1* |
| Address | 1Department of Microbiology, College of Medicine, The Catholic University of Korea, Seoul 137-701, Republic of Korea, 2Department of Food and Animal Biotechnology, Department of Agricultural Biotechnology, Center for Agricultural Biomaterials, and Research Institute for Agriculture and Life Sciences, Seoul National University, Seoul 151-921, Republic of Korea |
| Bibliography | Journal of Microbiology, 51(5),671-675, 2013, |
| DOI | 10.1007/s12275-013-3199-1 |
| Key Words | viral gastroenteritis, multiplex, Real-time PCR, RT-PCR, PCR |
| Abstract | Norovirus, Rotavirus group A, the Hepatitis A virus, and Coxsackievirus are all common causes of gastroenteritis. Conventional diagnoses of these causative agents are based on antigen detection and electron microscopy. To improve the diagnostic potential for viral gastroenteritis, internally controlled multiplex real-time polymerase chain reaction (PCR) methods have been recently developed. In this study, individual real-time PCRs were developed and optimized for specific detections of Norovirus genogroup I, Norovirus genogroup II, Rotavirus group A, the Hepatitis A virus, and Coxsackievirus group B1. Subsequently, individual PCRs were combined with multiplex PCR reactions. In general, multiplex real-time PCR assays showed comparable sensitivities and specificities with individual assays. A retrospective clinical evaluation showed increased pathogen detection in 29% of samples using conventional PCR methods. Prospective clinical evaluations were detected in 123 of the 227 (54%) total samples used in the multiplex realtime PCR analysis. The Norovirus genogroup II was found most frequently (23%), followed by Rotavirus (20%), the Hepatitis A virus (4.5%), Coxsackievirus (3.5%), and Norovirus genogroup I (2.6%). Internally controlled multiplex real-time PCR assays for the simultaneous detection of Rotavirus, Coxsackievirus group B, the Hepatitis A virus, and Norovirus genogroups I and II showed significant improvement in the diagnosis of viral gastroenteritis. |