Korean J Vet Res.  2012 Jun;52(2):75-82.

Development of a multiplex PCR to identify Salmonella, Leptospira and Brucella species in tissue samples

• Lam TQ
• Yoon BI
• Hahn TW

Abstract

We have developed and optimized a multiplex polymerase chain reaction (mPCR) for simultaneous detection of Brucella, Salmonella and Leptospira with high sensitivity and specificity. Three pairs 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 not produce any nonspecific amplification products when tested against 15 related species of bacteria. The sensitivity 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 examined by mPCR. The high specificity and sensitivity of this mPCR assay provide a valuable tool for diagnosis and for the simultaneous and rapid detection of three zoonotic bacteria that cause disease in both humans and animals. Therefore, this assay could be a useful alternative to the conventional method of culture and single PCR for the detection of each pathogen.

Keyword

Brucella; Leptospira; Salmonella species; multiplex PCR; tissue samples

MeSH Terms

Animals
Bacteria
Brucella
Cats
DNA Primers
Humans
Kidney
Leptospira
Liver
Multiplex Polymerase Chain Reaction
Polymerase Chain Reaction
Rats
Salmonella
Sensitivity and Specificity
Spleen
DNA Primers