PCR法检测人类脲原体U.Parvum和U.Urealyticum  

PCR assays for differential detection of Ureaplasma parvum and Ureaplasma urealyticum

作　　者：张新[1] 蒲晓允[1] 

机构地区：[1]第三军医大学新桥医院检验科,重庆400037

出　　处：《第三军医大学学报》2006年第17期1784-1787,共4页Journal of Third Military Medical University

摘　　要：目的建立2个快速、同步检测脲原体U.Parvum和U.Urealyticum的PCR反应体系。方法根据脲原体尿素酶B基因序列,设计2对分别针对U.Parvum和U.Urealyticum的特异性引物,建立2个PCR反应体系。通过脲原体标准株和临床阳性标本的PCR产物测序以及进行特异性和敏感性实验,对2个PCR体系进行评价。应用2个PCR反应体系对48例临床标本进行脲原体检测以及与培养法检测脲原体进行方法学比较。结果2个PCR体系能够检测U.Par-vum、U.Urealyticum标准株和临床标本中脲原体,扩增产物长度与目的片段大小一致。标准株和2个临床阳性标本扩增产物测序结果与GenBank中公布的序列完全一致。2个PCR体系与其他14株细菌(包括5株有尿素酶细菌)无交叉反应,检测限度均为10拷贝/ul的重组质粒DNA。对48例临床标本同时进行PCR和培养法脲原体检测,PCR的阳性检出率为54.2%,培养法为39.6%(P<0.05)。与培养法比较,PCR的灵敏度为94.7%,而与PCR法相比,培养法的灵敏度为69.2%。结论2个PCR反应体系能够快速、同步进行临床标本中脲原体的分种鉴别,具有高度特异性和灵敏度。Objective To establish two PCR assays to rapidly and simultaneously detect U. parvum and U. urealyticum. Methods Two PCR assays were established through designing two sets of specific primer targeting urease gene B of U. parvum and U. urealyticum, respectively. The standard strains of U. parvum and U. urealyticum and the clinical samples were detected by these PCR assays and PCR products of the standard strains and two clinical specimens were directly sequenced and carried out specific and sensitive assays. Fortyeight clinical specimens were tested for culture and the PCR assays and tow methods were compared. Results The standard strains of U. parvum, U. urealyticum and the clinical specimens were successfully and differentially detected by the PCR assays and the sequencing results were found to have a complete similarity to the Gen- Bank sequences. The 14 strains of other bacterial genera including 5 strains that produced urease were not amplified by these PCR assays. Each assay had a detection limit of 10 copies/μl of plasmid DNA. The positive rate in 48 clinical specimens by PCR （54.2%） was higher than that by culture （39.6%） （P 〈0.05 ）. The sensitivity of PCR was 94.5% and that of culture was 69.2%. Conclusion Tow PCR assays were successfully developed for the rapid and simultaneous detection of U. parvum and U. urealyticum in the clinical specimens and they are both specific and sensitive.

关 键 词：U.Parvum U. UREALYTICUM PCR 

分 类 号：R375[医药卫生—病原生物学] R446.9[医药卫生—基础医学]