Ann Clin Microbiol.  2013 Mar;16(1):33-38. 10.5145/ACM.2013.16.1.33.

Detection of 13 Enteric Bacteria and 5 Viruses Causing Acute Infectious Diarrhea Using Multiplex PCR from Direct Stool Specimens

Affiliations
  • 1Department of Laboratory Medicine, Incehon St. Mary's Hospital, The Catholic University School of Medicine, Incheon, Korea.
  • 2Department of Laboratory Medicine, Seoul St. Mary's Hospital, The Catholic University School of Medicine, Seoul, Korea. yjpk@catholic.ac.kr
  • 3Department of Laboratory Medicine, Uijeongbu St. Mary's Hospital, The Catholic University School of Medicine, Uijeongbu, Korea.
  • 4Department of Laboratory Medicine, St. Vincent's Hospital, The Catholic University School of Medicine, Suwon, Korea.
  • 5Department of Laboratory Medicine, Daejeon St. Mary's Hospital, The Catholic University School of Medicine, Daejeon, Korea.
  • 6Department of Laboratory Medicine, Yeouido St. Mary's Hospital, The Catholic University School of Medicine, Seoul, Korea.

Abstract

BACKGROUND
We investigated the prevalence of various pathogens (13 enteric bacteria and 5 viruses) which cause diarrhea using multiplex PCR of stool specimens and compared two multiplex PCR methods for detecting diarrheagenic Escherichia coli.
METHODS
A total of 405 stool specimens submitted between November 2010 to February 2011 for routine culture of enteric pathogens were included and screened for five viruses (astrovirus, Group A rotavirus, enteric adenovirus, norovirus G1/G2) and eight bacteria (Salmonella spp., Shigella spp., Campylobacter spp., Vibrio spp., C. difficile Toxin B, C. perfringens, Y. enterolytica, Aeromonas spp.) using the Seeplex(R) Diarrhea ACE detection kit (Seegene). In addition, virulence-associated genes of enteropathogenic E. coli, (EPEC), enterohemorrhagic E. coli (EHEC), enteroinvasive E. coli, (EIEC), enterotoxigenic E. coli (ETEC), and enteroaggressive E. coli (EAEC) were detected using 16-plex PCR and a commercial diarrheagenic E. coli detection (DEC) PCR kit (SSI Diagnostica).
RESULTS
Overall, 138 (34.1%) of 405 samples was positive for pathogen. The positive rate for virus was 18.5%. norovirus G2, Group A rotavirus, enteric adenovirus, astrovirus and norovirus G1 were detected in 40, 23, 8, 3 and 1 samples, respectively. The positive rate for bacteria was 24.4% (99/405). C. difficile toxin B was the most frequently detected, followed by C. perfringens, EPEC, and EAEC. The agreements of the two multiplex PCR methods for detecting EPEC and EHEC were 99.3% and 100%, respectively.
CONCLUSION
The detection rate was high (34.1%) including various diarrheagenic E. coli (6.2%) and C. perfringens (5.2%). Multiplex PCR is thus useful for detecting various pathogens.

Keyword

Agreement; Diarrheagenic Escherichia coli; Norovirus; Rotavirus
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