Development of PCR assays for species- and type-specific identification of Pasteurella multocida isolates
Development of PCR assays for species- and type-specific identification of Pasteurella multocida isolates
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DOI：
10.1128/jcm.36.4.1096-1100.1998
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发表时间：
1998-04-01
影响因子：
9.4
通讯作者：
Dawkins, HJS
中科院分区：
文献类型：
--
作者：
Townsend, KM;Frost, AJ;Dawkins, HJS
密切相关的多杀性巴氏杆菌分离株的基因组消减杂交产生了用于区分出血性败血症引起的B型菌株与其他多杀性巴氏杆菌血清型的克隆。在这些克隆的测序过程中设计的寡核苷酸引物已被证明在开发用于多杀性巴氏杆菌（特别是B：2型）的快速种特异性和型特异性检测的PCR测定中是有价值的。本研究证明，根据克隆6 B（KTT 72和KTSP 61）的序列设计的引物对特异性扩增了B：2、B：5和B：2，5型多杀性巴氏杆菌的DNA片段，引物KMT 1T7和KMT 1SP 6产生了所有分析的多杀性巴氏杆菌分离株特有的扩增产物。研究还表明，直接对细菌菌落或培养物进行PCR扩增是一种极其快速、灵敏的多杀性巴氏杆菌鉴定方法。
Genomic subtractive hybridization of closely related Pasteurella multocida isolates has generated clones useful in distinguishing hemorrhagic septicemia-causing type B strains from other P. multocida serotypes. Oligonucleotide primers designed during the sequencing of these clones have proved valuable in the development of PCR assays for rapid species-and type-specific detection of P. multocida and of type B:2 in particular. This study demonstrated that the primer pair designed from the sequence of the clone 6b (KTT72 and KTSP61) specifically amplified a DNA fragment from types B:2, B:5, and B:2,5 P. multocida and that the primers KMT1T7 and KMT1SP6 produced an amplification product unique to all P. multocida isolates analyzed. It was also shown that PCR amplification performed directly on bacterial colonies or cultures represents an extremely rapid, sensitive method of P. multocida identification.