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Animal Bioresource in Japan

.: Home > Animal Bioresource in Japan > 2013 > Volume 62 Number 1 > Eui-Suk JEONG, Kyoung-Sun LEE, Seung-Ho HEO, Jin-Hee SEO, Yang-Kyu CHOI

Rapid Identification of Klebsiella pneumoniae, Corynebacterium kutscheri, and Streptococcus pneumoniae Using Triplex Polymerase Chain Reaction in Rodents

Eui-Suk JEONG, Kyoung-Sun LEE, Seung-Ho HEO, Jin-Hee SEO, Yang-Kyu CHOI
Department of Laboratory Animal Medicine, College of Veterinary Medicine, Konkuk University, 1 Hwayang-dong, Gwangjin-gu, Seoul 143-701, Republic of Korea
Abstract :

Klebsiella pneumoniae, Corynebacterium kutscheri, and Streptococcus pneumoniae are important pathogens that cause respiratory infections in laboratory rodents. In this study, we used species-specific triplex PCR analysis to directly detect three common bacterial pathogens associated with respiratory diseases. Specific targets were amplified with conventional PCR using the tyrB gene from K. pneumoniae, gyrB gene from C. kutscheri, and ply gene from S. pneumoniae. Our primers were tested against purified DNA from another eleven murine bacteria to determine primer specificity. Under optimal PCR conditions, the triplex assay simultaneously yielded a 931 bp product from K. pneumoniae, a 540 bp product from C. kutscheri, and a 354 bp product from S. pneumoniae. The triplex assay detection thresholds for pure cultures were 10 pg for K. pneumoniae and S. pneumoniae, and 100 pg for C. kutscheri. All three bacteria were successfully identified in the trachea and lung of experimentally infected mice at the same time. Our triplex PCR method can be used as a useful method for detecting pathogenic bacterial infections in laboratory rodents.

Keywords :
Corynebacterium kutscheri , Klebsiella pneumoniae , Streptococcus pneumoniae , triplex PCR

Date Deposited : 06 Apr 2015 11:13

Last Modified : 06 Apr 2015 11:13

Official URL: http://https://www.jstage.jst.go.jp/browse/expanim/62/1/_contents

Volume 62, Number 1, January 2013

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