Abstract
College of Veterinary Medicine and Institute of Veterinary Science, Kangwon National University, Chuncheon 200-701, Korea(Received: August 4, 2011; Revised: January 3, 2012; Accepted: January 5, 2012)Abstract : We have developed and optimized a multiplex polymerase chain reaction (mPCR) forsimultaneous detection of Brucella, Salmonella and Leptospira with high sensitivity and specificity. Threepairs of oligonucleotide primers were designed to specifically amplify the targeted genes of Salmonella,Leptospira and Brucella species with sizes of 521, 408 and 223 bp, respectively. The mPCR did notproduce any nonspecific amplification products when tested against 15 related species of bacteria. Thesensitivity of the mPCR was 100 fg for Brucella and 1 pg for both Salmonella and Leptospira species.In the field application, kidney, liver and spleen were collected from wild rats and stray cats and examinedby mPCR. The high specificity and sensitivity of this mPCR assay provide a valuable tool for diagnosisand for the simultaneous and rapid detection of three zoonotic bacteria that cause disease in both humansand animals. Therefore, this assay could be a useful alternative to the conventional method of cultureand single PCR for the detection of each pathogen.Keywords : Brucella, Leptospira, Salmonella species, multiplex PCR, tissue samples
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