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

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pISSN 2288-0585 eISSN 2288-6850

Direct Application of Multiplex PCR on Stool Specimens for Detection of Enteropathogenic Bacteria

Original article

Annals of Clinical Microbiology (Ann Clin Microbiol) 2010 December, Volume 13, Issue 4, pages 162-168.

https://doi.org/10.5145/ACM.2010.13.4.162

Direct Application of Multiplex PCR on Stool Specimens for Detection of Enteropathogenic Bacteria

Min-Chul Cho1, Sin-Ae Noh1, Mi-Na Kim1, Kyoung-Mo Kim2
Departments of 1Laboratory Medicine and 2Pediatrics, Asan Medical Center and University of Ulsan College of Medicine, Seoul, Korea

Abstract

Background: Causative bacterial agents of infectious diarrheal disease were traditionally diagnosed by stool cultures. Stool culture, however, has a problem because of relatively low sensitivity and long turnaround time. In this study, we evaluated multiplex PCR applied on stool specimens directly to diagnose enteropathogenic bacteria. 

Methods: From June to September 2009, 173 diarrheal stools submitted for stool cultures were tested by SeeplexⓇ Diarrhea ACE Detection kit (Seegene, Korea) to detect 10 enteropathogenic bacteria. Specimens were cultured for Salmonella, Shigella, Vibrio, and Yersinia. Late 50 specimens were also cultured for Campylobacter. The specimens positive for verotoxin-producing Escherichia coli (VTEC) were further subcultured for detecting enterohaemorrhagic Escherichia coli O157:H7. Electronic medical records were reviewed for clinical and laboratory findings. 

Results: Of 173 specimens, multiplex PCR and cultures identified enteropathogens in 36 (20.8%) and 8 specimens (4.6%), respectively. While multiplex PCR detected 5 Salmonella, 15 Campylobacter, 1 Vibrio, 4 Clostridium difficiles toxin B, 5 Clostridium perfringens, 1 Yersinia enterocolitica, 5 Aeromonas, and 2 VTEC, cultures detected 5 Salmonella, 1 Vibrio, 1 Y. enterocolitica, 1 Aeromonas, and 2 E. coli O157:H7. 

Conclusion: Multiplex PCR would be useful to detect Campylobacter, VTEC and C. perfringens, as well as have equivalent sensitivity to conventional culture for ordinary enteropathogens such as Salmonella, Shigella, Vibrio, Y. enterocolitica. Direct application of multiplex PCR combined with conventional cultures on stool warrants remarkable improvement of sensitivity to diagnose enteropathogenic bacteria. (Korean J Clin Microbiol 2010;13:162-168)

Keywords

 Multiplex PCR, Infectious diarrheal disease, Enteropathogen