Fisher L N, Mathiesen M R, Duhamel G E
Department of Veterinary and Biomedical Sciences, University of Nebraska-Lincoln, 68583-0905, USA.
Clin Diagn Lab Immunol. 1997 Nov;4(6):681-6. doi: 10.1128/cdli.4.6.681-686.1997.
Forty-one reference and field isolates of intestinal spirochetes representing Serpulina hyodysenteriae, Serpulina innocens, Serpulina pilosicoli, Brachyspira aalborgi, and nonclassified weakly beta-hemolytic intestinal spirochetes were compared by restriction fragment length polymorphism (RFLP) of the periplasmic flagellar (PF) flaA1 gene. Six genetically distinct groups (I through VI), each with a unique RFLP fingerprint pattern, were identified by Southern blotting analysis of EcoRV chromosomal DNA digests with a PCR-amplified digoxigenin-labeled 1-kb fragment of the S. hyodysenteriae isolate B78 PF flaA1 gene. The RFLP fingerprint patterns corresponded to known DNA homology differences between Serpulina species and to provisionally designated species described previously by using phenotypic and genotypic classification schemes. RFLP fingerprinting of the PF flaA1 gene provides a relatively simple genotypic method for identification of intestinal spirochetes without the use of radioisotopes.
通过对周质鞭毛(PF)flaA1基因进行限制性片段长度多态性(RFLP)分析,比较了41株代表猪痢疾蛇形螺旋体、无害蛇形螺旋体、结肠螺旋体、奥尔堡短螺旋体和未分类的弱β-溶血肠道螺旋体的参考菌株和现场分离株。用猪痢疾蛇形螺旋体分离株B78 PF flaA1基因的PCR扩增地高辛标记1 kb片段,对EcoRV染色体DNA酶切产物进行Southern印迹分析,鉴定出6个遗传上不同的组(I至VI),每组都有独特的RFLP指纹图谱。RFLP指纹图谱与蛇形螺旋体属之间已知的DNA同源性差异以及先前使用表型和基因型分类方案描述的暂定种相对应。PF flaA1基因的RFLP指纹图谱提供了一种相对简单的基因型方法,无需使用放射性同位素即可鉴定肠道螺旋体。