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

被引:18
作者
Jeong, Eui-Suk [1 ]
Lee, Kyoung-Sun [1 ,2 ]
Heo, Seung-Ho [1 ,3 ]
Seo, Jin-Hee [1 ]
Choi, Yang-Kyu [1 ]
机构
[1] Konkuk Univ, Coll Vet Med, Dept Lab Anim Med, Seoul 143701, South Korea
[2] Osong Med Innovat Fdn Lab Anim Ctr, Cheongwon Gun 363951, Chungbuk, South Korea
[3] Univ Ulsan, Coll Med, Asan Inst Life Sci, Seoul 138736, South Korea
关键词
Corynebacterium kutscheri; Klebsiella pneumoniae; Streptococcus pneumoniae; triplex PCR; PSEUDOMONAS-AERUGINOSA; LABORATORY MICE; RATS; PCR; INFECTIONS; PATHOGENS; SAMPLES; ASSAY;
D O I
10.1538/expanim.62.35
中图分类号
S85 [动物医学(兽医学)];
学科分类号
0906 ;
摘要
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.
引用
收藏
页码:35 / 40
页数:6
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