title: Simultaneous detection of waterborne viruses by multiplex real-time 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 real-time 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.
authors.affiliation: Department of Microbiology, College of Medicine, The Catholic University of Korea, Seoul, 137-701, Republic of Korea.
authors.forename: Lae-Hyung
authors.initials: LH
authors.lastname: Kang
authors.forename: Se-hwan
authors.initials: SH
authors.lastname: Oh
authors.forename: Jeong-Woong
authors.initials: JW
authors.lastname: Park
authors.forename: Yu-Jung
authors.initials: YJ
authors.lastname: Won
authors.forename: Sangryeol
authors.initials: S
authors.lastname: Ryu
authors.forename: Soon-Young
authors.initials: SY
authors.lastname: Paik
mesh_terms: D005517:Foodborne Diseases / Q000175:diagnosis* / Q000821:virology; D005759:Gastroenteritis / Q000175:diagnosis* / Q000821:virology; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D012328:RNA Viruses / Q000235:genetics / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014773:Virology / Q000379:methods*; D014871:Water Microbiology
references.citation: J Virol. 2004 Jul;78(14):7795-802
references.pmid: 15220453
references.citation: J Med Virol. 2009 Nov;81(11):1903-11
references.pmid: 19774686
references.citation: Eur J Clin Microbiol Infect Dis. 2007 May;26(5):311-23
references.pmid: 17447091
references.citation: Clin Diagn Virol. 1998 Apr;9(2-3):115-23
references.pmid: 9645993
references.citation: Lancet. 1998 May 30;351(9116):1643-9
references.pmid: 9620732
references.citation: J Infect Dis. 2002 Jul 1;186(1):1-7
references.pmid: 12089655
references.citation: J Clin Virol. 2010 Nov;49(3):205-10
references.pmid: 20829103
references.citation: J Med Virol. 2004 May;73(1):118-22
references.pmid: 15042658
references.citation: Am J Epidemiol. 2001 Oct 1;154(7):666-74
references.pmid: 11581101
references.citation: J Med Virol. 2007 May;79(5):582-90
references.pmid: 17385670
references.citation: Pediatr Infect Dis J. 1999 May;18(5):420-6
references.pmid: 10353514
references.citation: Vaccine. 2000 Feb 18;18 Suppl 1:S57-60
references.pmid: 10683550