摘要
作者选用钩体赖型56601株、017株、秋季型56606株、曼耗2型67020株、1987年分离株87112、87369株,用内切酶EcoR Ⅰ、Bgl Ⅱ、Hha Ⅰ和Hind Ⅲ消化分析其DNA,结果表明:钩体017株的四种内切酶图谱明显不同,在EcoR Ⅰ酶切图谱上,赖型、秋季型、曼耗2型钩体DNA的主要差别在高分子区带,赖型强毒力株017较同型弱毒力株56601在10.5kb处染色较深;三型钩体的Hind Ⅲ酶切图谱仅有细小差异;分离株87112和87369的EcoR I 酶切图谱与赖型56601和017基本一致。在HindⅢ酶切图谱上,分离株87112与赖型56601株、017株相似。提示:限制性核酸内切酶分析可以作为钩体分型和鉴定的手段。
For the application of res-triction endonuclease analysis in typing andidentifying leptospira, we selected someserovars and isolates, and analysed preli-minarily their DNA with four restrictionenzymes, EcoR Ⅰ, Bgl Ⅱ, Hha Ⅰ, andHind Ⅲ. The DNA samples were isolatedfrom the reference strains and isolates asfollows: Serovar lai 56601, 017 (the viru-lent strain for PDH model of guinea pig),Serovar autumnalis 56606, Serovar manhaoⅡ 67020, and isolates 87112 and 87369. Each2 μof DNA was digested with 20u of re-striction enzyme at 37℃ for 2h and elec-trophoresed in 0.8% agarose gel. The gelswere stained in ethidium bromide andphotographed with UVllight In our experi-ments, apparently different restrictionpatterns of serovar lai 017 were observedwith four restriction enzymes. Serovarlai, serovar autumnalis and serovar man-hao Ⅱ showed different patterns withEcoR Ⅰ, especially in high molecular re-gions. We also observed in serovar lai 017a distinct 10. 5kb band which was obscurein 56601, the reference strain of serovarlai, after EcoR Ⅰ digestion. The threeserovars showed some delicate differencesin Hind Ⅲ restriction pattern. The twoisolates from Apodemus agrarius in Sichuan(1987) 87112 and 87369 had patternsidentical to those of serovar lai 56601, 017with EcoR Ⅰ, and 87112 also had a patternidentical to 56601, 017 with Hind Ⅲ. Our results indicate that selectedthree serovars can be identified by analysisof their DNA with EcoR Ⅰ and Hind Ⅲ.It is suggested that restriction endonu-clease analysis be a good method in typingand indentifying leptospira and in studyingthe differences of special DNA molecules.
出处
《华西医科大学学报》
CAS
CSCD
1991年第3期249-252,共4页
Journal of West China University of Medical Sciences
