用PCR法检测致肾盂肾炎大肠杆菌papA基因  

Detection of Pap A Gene from Uropathogenic E. coli with Polymerase Chain Reaction

作　　者：陈锦英[1] 李方涛[1] 康宁[1] 李宝艳[1] 

机构地区：[1]天津医科大学微生物学教研室,300070

出　　处：《天津医药》2001年第3期160-162,T001,共4页Tianjin Medical Journal

基　　金：国家自然科学基金

摘　　要：目的：用PCR方法检测致肾盂肾炎大肠杆菌(UPEC)的papA基因，探索建立UPEC的鉴定方法。方法：根据已发表的F13型papA基因序列设计引物，采用PCR方法扩增700bp左右的papA基因，并与血凝试验相比较，用于不同血清型P菌毛菌株和尿源性大肠杆菌的检测。结果：7个血清型P菌毛粘附基因群的papA PCR扩增均出现阳性结果，阴性对照株均无非特异性反应。39株尿源性大肠杆菌中MRHA阳性菌株的检出率为74.4％，papA PCR扩增的阳性率为71.8％，两组结果无显著性差异(P＞0.05)。菌细胞悬液直接用于PCR检测的敏感性为3×105～3×106cfu/ml。结论：papAPCR方法用于UPEC菌株的鉴定有良好的应用前景。To detect pap A gene from uropathologic E. coli( UPEC) with polymerase chain reaction(PCR)in order to establish an assay to identify UPEC strains. Methods: A pair of oligonucleotide primers derived from the published DNA sequence of F13 serotype pap A were used in PCR to emplify about 700bp-long pap A gene. As compared with hemagglutina-tion test,PCR method was applied to detect pap A gene of the different serotype P-piliated strains as well as a collection of 39 E. coli strains isolated from urine of patients with urinary tract infections. Results: Seven serotype pap A genes were all positive, while sepecific amplification in negative control strains were not found in PCR detection. The detection rates of MRHA and papA PCR positive strains in the tested isolates were 74.4 % and 71.8%, respectively. There was no statistical significance between the above results(P> 0.05). When bacterial cells were used directly in PCR detection, a sensitivity of 3 X 105 - 106 cfu/ml was demonstrated. Conclusion: The pap A PCR method may have a good application prospect in the identification of UPEC strains.

关 键 词：大肠杆菌 肾盂肾炎 聚合酶链反应 papA基因 

分 类 号：R692.7[医药卫生—泌尿科学]