摘要
目的检测青藏铁路沿线鼠疫菌表型性状、质粒组成、外膜蛋白种类及对抗生素的敏感性。方法采用常规糖醇发酵管法检测生化等表型性状;采用反向血凝试验、用假结核菌为复盖种子层法、用草酸镁培养基和刚果红色素培养基检测FraI、Pstl、Vwa和Pgm4种毒力因子;用Lawton最小培养基测定营养型;采用常规的动物接种试验测定毒力;采用碱裂解、酚一氯仿抽提法提取鼠疫菌质粒;鼠疫菌经破碎后采取超高速离心法提取外膜蛋白;采用纸片法进行抗生素敏感性试验。结果实验用的青藏铁路沿线鼠疫菌株生化性状表现为甘油+、鼠李糖一、阿胶糖+、密二糖一、麦芽糖+、脱氮+的特征;具有鼠疫菌荚膜抗原(FraI+)、毒力抗原(Vwa+)、鼠疫菌素I(PstI+)、色素沉着因子(Pgm+)4种毒力因子;营养型表现为对Trp、Thr、Leu、Arg不依赖,而对Phe、Met、Cys依赖,对Ile和Glu表现出低营养;对小白鼠的LD100在10^2~10^3个菌,LDs。为7.94-31.62个菌,对豚鼠的LD100在10^5~10^7个菌,LD50为68.14-685.49个菌;携带Mr为6×10^6、45×10^6、52×10^6、65×10^6、92×10^6]质粒;在10%的SDS-PAGE上37℃条件下的培养物可表达26条外膜蛋白带,28℃培养物可表达36条外膜蛋白;部分头孢类和喹诺酮类药物对实验菌株的敏感性效果较好,20种抗生素均较链霉素为佳。结论青藏铁路沿线的鼠疫菌属于典型的青藏高原型菌株,都是强毒力鼠疫菌,临床治疗除常规的链霉素外可选用头孢类和喹诺酮类药物,该项研究可为青藏铁路沿线的鼠疫防治提供科学的依据。
Objective To study and detect the character and plasmid and outer membrane proteins and antibi- otic sensitivity of the Yersinia pestis along the Qinghai-Tibet Railway. Methods Some phenotype characters of sug- ar-alcohol fermentation was adopted. RIHA, seed layer method by the Yersinia pseudotuberculosis,magnesium ox- alate medium and congo red medium were adopted to detect FraI, Pstl, Vwa and Pgm; Lawton medium was adopted to detect nutritional type;animal inoculation test was adopted to detect virulence; alkaline lysis, Phenol-chloroform extraction was adopted to distill the plasmid of the Y. pestis; ultracentrifuge was adopted to extract outer membrane proteins of Yersinia pestis ; scrip method was adopted to test the antibiotic susceptibility. Results The Y. pestis bi- ochemistry character included glycerol +, rhamnose --, melibiose --, maltose +, denitrification + ; there were FraI +, Vwa +, Pst I +, Pgm in animal experiment the white mouse LD100 : 10^2-10^3, LD50 : 7.94~31.62, the cavy LD100 : 10^5 -10^7, LDs0 : 68.14-685.49; the Yersinia pestis along Qinghai--Tibet Railway depended on Phe, Met, Cys, not on Trp, Thr, Leu, Arg; the tests showed meiotroph of Ile and Glu, and contained the plasmid of 6 ×10^6 、45×10^6、 52×10^6、65×10^6、 92 ×10^6Mr; therewere 26 stripes of outer membrane proteins at 37℃ and 36 stripes at 28℃ in 10% SDS-PAGE eleetrophoresis; the Yersinia pestis was sensitive to some of cephalosporin and carbostyril, and 20 antibiotics were better than streptomycin. Conclusion the Yersinia pestis along Qinghai-Tibet Railway is standard strain of Qinghai-Tibet plateau ecotype. The research might provide scientific evidence for pre- vention and control of plague along the Qinghai-Tibet Railways.
出处
《国外医学(医学地理分册)》
CAS
2009年第3期108-112,共5页
Foreign Medical Sciences:Section of Medgeography
基金
"十五"国家科技攻关资助项目(No.2004BA718B08)
关键词
青藏铁路沿线
鼠疫菌
表型性状
质粒
外膜蛋白
抗生素敏感性
along the Qinghai-Tibet Railways
the Yersinia pestis
phenotype character
plasmid
the outer membrane proteins
antibiotic sensitivity