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Title: MULTIPLEX POLYMERASE CHAIN REACTION FOR THE RAPID DETECTION AND IDENTIFICATION OF CAMPYLOBACTER AND LISTERIA

Author
item HARMON, KAREN - IOWA STATE UNIV., AMES
item Wesley, Irene

Submitted to: North Central Conference of Veterinary Laboratory Diagnosticians
Publication Type: Abstract Only
Publication Acceptance Date: 6/14/1995
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: The detection and identification of zoonotic pathogens is often a laborious and time-consuming process. Improved diagnostic techniques are continually being sought. Campylobacter, a cause of intestinal disease as well as abortion in domestic animals, is one microorganism of concern. The methods commonly used for Campylobacter involve lengthy incubations and specialized gas mixtures. Additionally, tests to differentiate between C. coli and C. jejuni are not always reliable. A multiplex polymerase chain reaction (mPCR) assay was developed for the detection of Campylobacter from swine rectal swabs. The assay can detect C. coli/jejuni and differentiate between the 2 species. Two sets of primers are used in this assay. The first set amplifies a portion of the flagellin genes of both C. coli and C. jejuni. The second set of primers amplifies a DNA target found only in C. jejuni. Based on the band pattern resulting from electrophoresis of the mPCR product, one can determine the presence of either C. coli or C. jejuni and also differentiate between these two organisms. Another causative agent of abortion in domestic animals is Listeria monocytogenes. Traditional methods for the isolation and identification of L. monocytogenes require specialized media and procedures. A mPCR assay was developed for Listeria, enabling us to rapidly determine the presence of Listeria and, simultaneously, confirm whether the Listeria present is L. monocytogenes. Two sets of primers are used. One amplifies a segment in all Listeria tested; the other targets a portion of DNA found only in L. monocytogenes. Like the Campylobacter mPCR assay, the electrophoretic banding pattern is indicative of the organism(s) present in the test sample.