PCR for the detection of Streptobacillus moniliformis
PCR for the detection of Streptobacillus moniliformis
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DOI：
10.1258/0023677021912352
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发表时间：
2002-04-01
影响因子：
2.4
通讯作者：
Thuis, HCW
中科院分区：
文献类型：
--
作者：
Boot, R;Oosterhuis, A;Thuis, HCW
为了研究聚合酶链反应（PCR）检测念珠状链杆菌的方法，根据11株念珠状链杆菌（Streptobacillusmoniliformis）16 SrRNA基因的部分DNA碱基序列设计了一对PCR引物。念珠状菌株PCR检测到的S.念珠状DNA从S.念珠状菌来自啮齿动物、人类和火鸡的菌株。从产坏死梭杆菌和白蚁Sebaldella（拟杆菌）中获得了大约相同大小的扩增子，但Bfa I处理这些扩增子没有导致S.念珠藻特异的130 bp DNA片段。14种其他梭杆菌属的计算机模拟评价。12个不可培养的植原体表明，没有一个可能产生混淆的扩增子或DNA片段。PCR检测到S.在所有四只经口和四只静脉内感染的C57 BL/6小鼠中均检测到念珠菌感染，并且从除一只小鼠之外的所有小鼠中培养细菌。PCR检测到S.在所有12只经口感染的WU大鼠中，以及在暴露于自然感染的8只大鼠中的5只中，均观察到念珠菌感染。酶联免疫吸附试验（ELISA）和PCR同样成功地检测感染大鼠，但S。培养未检出念珠状菌。
Streptobacillus moniliformis is a Gram-negative bacterium found in various laboratory animal species and is the cause of rat bite fever and Haverhill fever in man. In order to evaluate a polymerase chain reaction (PCR) for the detection of this zoonotic bacterium in animal tissues a set of primers was designed based on the DNA base sequence of part of the 16S rRNA gene from 11 S. moniliformis strains. The PCR detected as few as 2 6 copies of S. moniliformis DNA. A 296 bp DNA fragment was amplified from S. moniliformis Strains from rodents, humans and turkeys. Amplicons of about the same size were obtained from Fusobacterium necrogenes and Sebaldella (Bacteroidcs) termitidis but Bfa I treatment of these amplicons did not result in the S. moniliformis specific 130 bp DNA fragment. The in silico evaluation of 14 additional Fusobacterium spp. and 12 unculturable phytoplasmas indicated that none is likely to give rise to confusing amplicons or DNA fragments. The PCR detected S. moniliformis infection in all four orally- and four intravenously-infected C57BL/6 mice and the bacterium was cultured from all but one mouse. The PCR detected S. moniliformis infection in all 12 orally-infected WU rats, and in five of eight rats exposed to natural infection. Enzyme linked immunosorbent assay (ELISA) and PCR were equally successful in detecting infection in rats but S. moniliformis was not detected by using culture.