摘要
目的 研究临床分离的致肾盂肾炎大肠杆菌粘附基因群DNA的多态性。方法 以血凝试验 (MRHA)为基础 ,分别用编码P菌毛主体结构和尖端粘附素的基因探针 ,采用DNA杂交方法对临床分离的 95株尿源性大肠杆菌的粘附基因群进行分析。结果MRHA +++～ ++++占 37 9% (36株 ) ,++占 32 6 % (31株 ) ,MRHA阴性占 2 9 5 % (2 8株 )。菌落原位杂交和DNA斑点杂交试验在MRHA +++～ ++++,++和阴性菌株中两个探针均为阳性的分别为 91 7%、9 7%和 2 1 4% ;在MRHA ++和阴性菌株中单一探针阳性的分别为 6 4 5 %和 35 7%。同时 ,选 10株MRHA ++++大肠杆菌提取染色体DNA ,经EcoRⅠ酶切后Southern杂交进行限制性片段长度多态性分析 ,同源性DNA片段大小不一 ,杂交信号的强弱程度也不同。结论 提示致肾盂肾炎大肠杆菌pap粘附基因群的多样性。
Aim In order to study the DNA polymorphism of the adherence gene cluster among uropathogenic E coli (UPEC) strains Methods Based on the hemoagglutination test, the adherence gene cluster analyses were carried out by using DNA hybridization methods with the probes of the pap ACDH including the structural gene for the P-fimbriae subunit and the papEFG including the actual P-adhesin for 95 stains of E coli isolated from urine Results It was showed that MRHA ～,and negative strains were 37 9%,32 6% and 29 5%, respectively Using colony hybridization in situ and DNA dot blotting, the hybridized positive strains with above two probes among MRHA ～, and negative stains were 91 7%, 9 7% and 21 4%, respectively; the hybridized positive strains with one of both probes in MRHA and negative examined strains were 64 5% and 35 7% Meanwhile, the chromosomal DNA was extracted from the 10 strains with MRHA character and was digested with the restriction endonuclease EcoR Ⅰ Then the restriction fragment length polymorphism (RFLP) of UPEC were studied by Southern hybridization methods The homologous fragments of the tested strains with two probes were different in the size and the light and shade degree Conclusion It was inferred that pap adherence gene cluster of uropathogenic E coli strains may be very diversity
出处
《中国人兽共患病杂志》
CAS
CSCD
北大核心
2000年第3期15-17,共3页
Chinese Journal of Zoonoses
基金
国家自然科学基金
关键词
肾盂肾炎
大肠杆菌
粘附基因群
DNA杂交
多态性
Uropathogenic E coli
Adherence gene cluster
hemoagglutination
DNA hybridization
RFLP
