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United States Department of Agriculture

Agricultural Research Service

Research Project: EPIDEMIOLOGY, ECOLOGY, AND MOLECULAR GENETICS OF ANTIMICROBIAL RESISTANCE IN PATHOGENIC AND COMMENSAL BACTERIA FROM FOOD ANIMALS Title: A Multiplex Pcr Method for the Rapid Serotyping of Common Clinical Isolates of Salmonella Enterica Subspecies Enterica

Authors
item Boyle, David - STATE OF WASHINGTON
item Kim, Seonghan - STATE OF WASHINGTON
item Frye, Jonathan
item Gautom, Romesh - STATE OF WASHINGTON
item Hu, Jinxin - STATE OF WASHINTON
item Cray, Paula

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: March 23, 2006
Publication Date: March 23, 2006
Citation: Boyle, D.S., Kim, S., Frye, J.G., Gautom, R., Hu, J., Cray, P.J. 2006. A multiplex pcr method for the rapid serotyping of common clinical isolates of salmonella enterica subspecies enterica. Symposium on Molecular Serotyping and Subtyping of Enteric Bacteria. 03/23/2006. Atlanta, GA

Technical Abstract: Background: The bacterial species Salmonella enterica is one of the major causes of gastroenteritis in humans and has over 1,500 serotypes. Serotyping is the most common tool used to identify isolates from diseased patients. However, the serotyping method can take several weeks and sometimes can be impossible to perform. Presently, serotyping of Salmonella enterica is performed only at state and national reference laboratories because it requires skilled staff and requires a stock of many commercial antisera. In this study we demonstrate the potential of multiplex PCR to create an extremely rapid and accurate method to serotype the 31 most common serotypes of Salmonella enterica in WA State. Methods: Primers for two 5-plex PCR reactions were selected for target genes in the Salmonella enterica serovar Typhimurium LT2 and Salmonella enterica serovar typhi CT18 genomes. Results: A total of 253 specimens from 31 different serotypes including the top 20 clinical and top 10 veterinary serotypes were screened with the assay. From this data, unique amplification profiles for almost all of 31 Salmonella enterica serotypes were established. Other PCR assays were developed for further discrimination of similar serotypes when necessary. From 110 isolates screened by blind testing 102 isolates were correctly typed using this method. This is comparable with serotyping. Conclusions: Here we demonstrate a very rapid and simple molecular method for serotyping common Salmonella enterica serotypes. The time for serotyping is dramatically reduced to only 5 hours. The method is basic and does not need specialized staff and a large collection of antisera. The assay may be applied in any clinical facility which has PCR and electrophoresis equipment.

Last Modified: 10/21/2014
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