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United States Department of Agriculture

Agricultural Research Service

Title: Comparison of Methods for Differentiation of Salmonella Enterica Serovar Enteritidis Phage Type 4 Isolates

Authors
item Lopes, V - UNIV OF MN - ST PAUL, MN
item Velayundhan, B - UNIV OF MN - ST PAUL, MN
item Halvorson, D - UNIV OF MN - ST PAUL, MN
item Lauer, D - MN PLTRY TESTING LAB- MN
item GAST, RICHARD
item Nagaraja, K - UNIV OF MN - ST PAUL, MN

Submitted to: American Journal of Veterinary Research
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: November 14, 2003
Publication Date: May 1, 2004
Citation: Lopes, V.C., Velayundhan, B.T., Halvorson, D.A., Lauer, D.C., Gast, R.K., Nagaraja, K.V. 2004. Comparison Of Methods For Differentiation Of Salmonella Enterica Serovar Enteritidis Phage Type 4 Isolates. American Journal of Veterinary Research v.65 p.538-543 2004.

Interpretive Summary: Salmonella enteritidis is an important food-borne pathogen for humans. In order to accurately identify the sources of S. enteritidis that contribute to disease outbreaks, it is important to have methods for differentiating strains of this organism based on their molecular genetic properties. The present study evaluated an assortment of molecular typing methods for their ability to distinguish between a set of 27 S. enteritidis phage type 4 strains isolated in the United States and Europe. Of the six methods tested, random amplification of polymorphic DNA (RAPD) provided the greatest degree of discrimination between the various S. enteritidis strains, allowing the grouping of similar strains into clusters. This demonstrates that the RAPD method can be a valuable tool for epidemiological investigations.

Technical Abstract: OBJECTIVE - To compare molecular typing methods for the discrimination of Salmonella enterica serovar Enteritidis (S. enteritidis) phage type (PT) 4 isolates, allowing for the determination of their genetic relatedness. SAMPLE POPULATION ' Twenty seven S. enteritidis PT 4 strains isolated in the United States and Europe were used in this study. PROCEDURE - Several molecular typing methods were performed to assess their ability to genetically discriminate S. enteritidis PT 4 isolates. The results of pulse-field gel electrophoresis (PFGE), Repetitive-Polymerase Chain Reaction (PCR), 16S rRNA gene sequencing, random amplification of polymorphic DNA (RAPD), PCR-restriction fragment length polymorphism of 16S rRNA and antimicrobial sensitivity were evaluated. RESULTS - Compared to other techniques evaluated in this study, RAPD typing method with RAPD1 primer revealed to be the most discriminatory fingerprinting technique, allowing for clustering of S. enteritidis PT 4 isolates by their genetic similarity. CONCLUSION AND CLINICAL RELEVANCE- This study demonstrates the value of RAPD with RAPD1 primer as a tool for epidemiological investigations of S. enteritidis PT 4. It can be used complementarily with PFGE and phage typing to determine genetic relatedness of S. enteritidis isolates involved in outbreaks. A reliable and highly discriminatory method for epidemiological investigations is critical in order to identify the source(s) of infections and, consequently, prevent the spread of outbreaks of S. enteritidis PT 4.

Last Modified: 9/10/2014
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