Journal List > Korean J Clin Microbiol > v.13(4) > 1038201

Korean J Clin Microbiol. 2010 Dec;13(4):162-168. Korean.
Published online December 27, 2010.  https://doi.org/10.5145/KJCM.2010.13.4.162
Copyright © 2010 The Korean Society of Clinical Microbiology
Direct Application of Multiplex PCR on Stool Specimens for Detection of Enteropathogenic Bacteria
Min-Chul Cho,1 Sin-Ae Noh,1 Mi-Na Kim,1 and Kyoung-Mo Kim2
1Department of Laboratory Medicine, Asan Medical Center and University of Ulsan College of Medicine, Seoul, Korea.
2Department of Pediatrics, Asan Medical Center and University of Ulsan College of Medicine, Seoul, Korea.

Correspondence: Mi-Na Kim, Department of Laboratory Medicine, Asan Medical Center and University of Ulsan College of Medicine, 388-1 Pungnap 2-dong, Songpa-gu, Seoul 138-736, Korea. (Tel) 82-2-3010- 4511, (Fax) 82-2-478-0884, Email: mnkim@amc.seoul.kr
Received May 17, 2010; Revised August 05, 2010; Accepted September 10, 2010.

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.

Keywords: Multiplex PCR; Infectious diarrheal disease; Enteropathogen

Tables


Table 1
Target genes and their product size of multiplex PCR
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Table 2
Distribution of diarrhea pathogens confirmed by multiplex PCR and culture results
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Table 3
Clinical characteristics of PCR-positive patients
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