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通过限制性内切酶分析和Southern印迹分析鉴定和表征猪痢疾蛇形螺旋体。

Identification and characterization of Serpulina hyodysenteriae by restriction enzyme analysis and Southern blot analysis.

作者信息

Sotiropoulos C, Coloe P J, Smith S C

机构信息

Department of Applied Biology and Biotechnology, Royal Melbourne Institute of Technology University, Australia.

出版信息

J Clin Microbiol. 1994 May;32(5):1397-401. doi: 10.1128/jcm.32.5.1397-1401.1994.

Abstract

Chromosomal DNA restriction enzyme analysis and Southern blot hybridization were used to characterize Serpulina hyodysenteriae strains. When chromosomal DNAs from selected strains (reference serotypes) of S. hyodysenteriae were digested with the restriction endonuclease Sau3A and hybridized with a 1.1-kb S. hyodysenteriae-specific DNA probe, a common 3-kb band was always detected in S. hyodysenteriae strains but was absent from Serpulina innocens strains. When the chromosomal DNA was digested with the restriction endonuclease Asp 700 and hybridized with two S. hyodysenteriae-specific DNA probes (0.75 and 1.1 kb of DNA), distinct hybridization patterns for each S. hyodysenteriae reference strain and the Australian isolate S. hyodysenteriae 5380 were detected. Neither the 1.1-kb nor the 0.75-kb DNA probe hybridized with Asp 700- or Sau3A-digested S. innocens chromosomal DNA. The presence of the 3-kb Sau3A DNA fragment in S. hyodysenteriae reference strains from diverse geographical locations shows that this fragment is conserved among S. hyodysenteriae strains and can be used as a species-specific marker. Restriction endonuclease analysis and Southern blot hybridization with these well-defined DNA probes are reliable and accurate methods for species-specific and strain-specific identification of S. hyodysenteriae.

摘要

采用染色体DNA限制酶分析和Southern印迹杂交技术对猪痢疾蛇形螺旋体菌株进行鉴定。当用限制性内切酶Sau3A消化猪痢疾蛇形螺旋体选定菌株(参考血清型)的染色体DNA,并与一个1.1kb的猪痢疾蛇形螺旋体特异性DNA探针杂交时,在猪痢疾蛇形螺旋体菌株中总能检测到一条3kb的共同条带,而无害蛇形螺旋体菌株中则没有。当用限制性内切酶Asp 700消化染色体DNA,并与两个猪痢疾蛇形螺旋体特异性DNA探针(0.75kb和1.1kb DNA)杂交时,检测到每个猪痢疾蛇形螺旋体参考菌株和澳大利亚分离株猪痢疾蛇形螺旋体5380的独特杂交模式。1.1kb和0.75kb的DNA探针均未与经Asp 700或Sau3A消化的无害蛇形螺旋体染色体DNA杂交。来自不同地理位置的猪痢疾蛇形螺旋体参考菌株中存在3kb的Sau3A DNA片段,表明该片段在猪痢疾蛇形螺旋体菌株中保守,可作为种特异性标记。用这些明确的DNA探针进行限制性内切酶分析和Southern印迹杂交是猪痢疾蛇形螺旋体种特异性和菌株特异性鉴定的可靠且准确的方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/53e2/263714/f4f01a3ef382/jcm00005-0278-a.jpg

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