NDLI: Real-time multiplex PCR for simultaneous detection of Campylobacter jejuni, Salmonella, Shigella and Yersinia species in fecal samples.

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Real-time multiplex PCR for simultaneous detection of Campylobacter jejuni, Salmonella, Shigella and Yersinia species in fecal samples.

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Real-time multiplex PCR for simultaneous detection of Campylobacter jejuni, Salmonella, Shigella and Yersinia species in fecal samples.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Wiemer, Dorothea Loderstaedt, Ulrike Von Wulffen, Hinrik Priesnitz, Simone Fischer, Marcellus Tannich, Egbert Hagen, Ralf Matthias
Description	Country affiliation: Germany Author Affiliation: Wiemer D ( Department for Tropical Medicine at the Bernhard Nocht Institute, Bundeswehr Hospital Hamburg, Germany. Wiemer@bni-hamburg.de)
Abstract	Diarrheal diseases due to notifiable bacterial infections require rapid diagnosis of the causative pathogens. To facilitate detection, a real-time multiplex PCR was developed that identifies common diarrhea-causing bacteria in fecal samples. On the basis of published sequence data, sets of primers and probes were designed that were specific for Campylobacter jejuni, Salmonella, Shigella/enteroinvasive Escherichia coli EIEC, and Yersinia species, suitable for use in a one-tube PCR assay. The assay was assessed using a list of 137 well-defined intestinal bacterial strains or isolates. Furthermore, 393 routine clinical stool samples were analyzed, and the results of real-time multiplex PCR were compared with those obtained by established microbiological methods. The PCR yielded results within 3h including DNA purification. No false-positive signals or cross-reactions were observed. The analytical sensitivity was 10(3)cfumL(-1) for Campylobacter jejuni, 10(4)cfumL(-1) for Salmonella, and 10(5)cfumL(-1) for Shigella/EIEC and Yersinia, respectively. Compared with culture, PCR detected 79 out of 81 Campylobacter jejuni (97.5%), 71 out of 74 Salmonella (96%), 8 out of 8 Shigella (100%), and 10 out of 10 Yersinia-positive (100%) clinical samples. In culture-negative samples (n=192), PCR additionally detected 2 Shigella, 1 Salmonella, and 5 Campylobacter jejuni infections. Thus, the new real-time multiplex PCR provides reliable results within a short time and might be useful as an additional diagnostic tool whenever time is important in the diagnosis of enteropathogenic bacteria.
File Format	HTM / HTML
ISSN	14384221
Issue Number	7
Volume Number	301
e-ISSN	16180607
Journal	International Journal of Medical Microbiology
Language	English
Publisher	Elsevier
Publisher Date	2011-11-01
Publisher Place	Germany
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Discipline Microbiology Bacteriological Techniques Methods Diarrhea Microbiology Enterobacteriaceae Infections Diagnosis Enterobacteriaceae Isolation & Purification Feces Multiplex Polymerase Chain Reaction Real-time Polymerase Chain Reaction Genetics Humans Sensitivity And Specificity Time Factors Evaluation Studies Journal Article
Content Type	Text
Resource Type	Article
Subject	Infectious Diseases Microbiology Microbiology (medical)

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