CAJ | 학술논문

目的研究致肾盂肾炎大肠杆菌(UPEC)132特异性片段R049在UPEC菌株和正常人粪便分离的大肠杆菌菌株中的分布情况.方法对20株UPEC和加株粪便分离大肠杆菌采用PCR方法扩增R049片段,对R049阳性菌株检测编码5种毒力因子的基因(papC、fimH、hly、aer、cnf1),并对其基因组DNA用Xba Ⅰ酶切,进行脉冲场凝胶电泳分析,继而用毛细管法将DNA片段转移至尼龙膜,利用地高辛标记的R049 ORF探针片段进行Southem杂交,分析杂交条带的分布特征.结果 20株UPEC中有8株(40%)、40株粪便分离的大肠杆菌中有3株(7.5%)R049片段扩增阳性,两组差异有统计学意义(P<0.01).R049阳性菌株中,R049片段与5种UPEC毒力因子密切相关.Southem杂交结果表明3株粪便分离大肠杆菌R049阳性杂交带分别为150 kb、15 kb与240 kb;8株UPEC中有6株阳性杂交带均为350 kb,其余2株分别为280 kb和25 kb.结论 UPEC132特异性片段R049在国内分离的UPEC菌株中具有共有性和聚集性分布的特征.
목적 연구치신우신염대장간균(UPEC)132특이성편단R049재UPEC균주화정상인분편분리적대장간균균주중적분포정황.방법 대20주UPEC화가주분편분리대장간균채용PCR방법확증R049편단,대R049양성균주검측편마5충독력인자적기인(papC、fimH、hly、aer、cnf1),병대기기인조DNA용Xba Ⅰ매절,진행맥충장응효전영분석,계이용모세관법장DNA편단전이지니룡막,이용지고신표기적R049 ORF탐침편단진행Southem잡교,분석잡교조대적분포특정.결과 20주UPEC중유8주(40%)、40주분편분리적대장간균중유3주(7.5%)R049편단확증양성,량조차이유통계학의의(P<0.01).R049양성균주중,R049편단여5충UPEC독력인자밀절상관.Southem잡교결과표명3주분편분리대장간균R049양성잡교대분별위150 kb、15 kb여240 kb;8주UPEC중유6주양성잡교대균위350 kb,기여2주분별위280 kb화25 kb.결론 UPEC132특이성편단R049재국내분리적UPEC균주중구유공유성화취집성분포적특정.
Objective To study the distribution of the specific fragment R049 of uropathogenic E. coli(UPEC) 132 in UPEC and fecal E. coli strains. Methods The specific fragment R049 was amplified by PCR from 20 UPEC strains and 40 fecal E. coli strains, and 5 genes encoding virulence factors (papC, fimH, hly, aer, cnf1) were detected from fragment R049 positive strains. Pulse field gel electronphoresis (PFGE) was applied for isolating the Xba Ⅰ restriction fragments of the genomes of fragment R049 positive strains, and then Southern blot was applied for analyzing the distribution features of the positive hybridization bands by digoxin-labeled R049 ORF probe. Results The specific fragment R049 was amplified from 8 of 20 UPEC strains (40%) and 3 of 40 fecal E. coli strains (7.5%), and statistics analysis showed significant difference (P<0.01). The specific fragment 11049 was closely related with 5 virulence factors of UPEC in the fragment R049 positive strains. Southern blot showed the sizes of positive bands were 150 kb, 15 kb and 240 kb in 3 fecal E. coli strains, 350 kb in 6 of 8 UPEC strains, and 280 kb and 25 kb in the rest two UPEC strains. Conclusion The specific fragment R049 of UPEC132 possessed the feature of clustering distribu-tion in domestic isolated UPEC strains.