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Animal Bioresource in Japan

.: Home > Animal Bioresource in Japan > 2011 > Volume 60 Number 1 > Eui-Suk JEONG1), Kyoung-Sun LEE1), Seung-Ho HEO1)2), Jin-Hee SEO1) and Yang-Kyu CHOI1)

Triplex PCR for the Simultaneous Detection of Pseudomonas aeruginosa, Helicobacter hepaticus, and Salmonella typhimurium

Eui-Suk JEONG1), Kyoung-Sun LEE1), Seung-Ho HEO1)2), Jin-Hee SEO1) and Yang-Kyu CHOI1)
1) Department of Laboratory Animal Medicine, College of Veterinary Medicine, Konkuk University 2) Asan Institute for Life Sciences, University of Ulsan College of Medicine
Abstract :

The accurate and economical diagnosis of pathogenic bacteria is necessary for the microbiological control of laboratory animals. In this study, we developed a triplex PCR method for the direct detection of three common gastroenteric bacteria, Pseudomonas aeruginosa, Helicobacter hepaticus, and Salmonella typhimurium. Targets were specifically amplified by conventional PCR assay using a genomic fragment from P. aeruginosa, 16S ribosomal RNA from H. hepaticus, and the invA gene from S. typhimurium. To investigate the specificity of our primers, they were tested against purified DNA from many other bacterial species. There were no amplification products from other bacteria. Under optimized conditions, the triplex assay simultaneously yielded a 726-bp product from P. aeruginosa, a 417-bp product from H. hepaticus, and a 246-bp product from S. typhimurium. The detection limits of this assay in pure culture were 10 pg for P. aeruginosa, and 0.1 pg for H. hepaticus and S. typhimurium. All three bacteria were successfully detected in the liver, cecum, and feces of experimentally infected mice. This method is a useful and convenient assay that allows the simultaneous identification of bacterial pathogens in mice. Our triplex method will be used to improve quality control in the detection of pathogenic bacterial infections in laboratory animal facilities.

Keywords :
H. hepaticus, P. aeruginosa, S. typhimurium, triplex PCR

Date Deposited : 21 Jun 2011 11:19

Last Modified : 21 Jun 2011 11:19

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Volume 60, Number 1, - 2011

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