CAJ | 학술논문

目的了解生殖支原体对大环内酯类抗生素耐药相关的23S rRNA基因突变情况.方法就诊于我院性病门诊的91例近期应用过大环内酯类药物治疗的持续性和复发性尿道炎患者,取尿道拭子和前段尿标本进行生殖支原体(Mg)、沙眼衣原体(Ct)、解脲脲原体(Uu)等病原体检测,筛选出单一生殖支原体阳性患者标本,应用套式PCR扩增与大环内酯类抗生素耐药性相关的23S rRNA基因Ⅴ区片段,扩增产物进行DNA测序,测得序列与美国国立生物信息中心已登记的生殖支原体标准株G37的相应基因序列比对.结果 91例的标本中,Mg阳性21例,Ct阳性18例(19.8％),Uu阳性10例,Mg与UU同时阳性2例,Ct与Uu同时阳性3例,Mg 、Ct和Uu检测均为阴性者37例.21例Mg阳性标本中,18例的标本成功扩增出23S rRNA基因Ⅴ区片段,除1例未发现基因突变外,其余17例均发现2058和2059位点突变,其中A2059G突变10例,A2058G突变5例,A2058T突变2例.结论 23S rRNA基因Ⅴ区片段基因突变可能与南京及周边地区Mg对大环内酯类药物耐药性相关.
목적 료해생식지원체대대배내지류항생소내약상관적23S rRNA기인돌변정황.방법 취진우아원성병문진적91례근기응용과대배내지류약물치료적지속성화복발성뇨도염환자,취뇨도식자화전단뇨표본진행생식지원체(Mg)、사안의원체(Ct)、해뇨뇨원체(Uu)등병원체검측,사선출단일생식지원체양성환자표본,응용투식PCR확증여대배내지류항생소내약성상관적23S rRNA기인Ⅴ구편단,확증산물진행DNA측서,측득서렬여미국국립생물신식중심이등기적생식지원체표준주G37적상응기인서렬비대.결과 91례적표본중,Mg양성21례,Ct양성18례(19.8％),Uu양성10례,Mg여UU동시양성2례,Ct여Uu동시양성3례,Mg 、Ct화Uu검측균위음성자37례.21례Mg양성표본중,18례적표본성공확증출23S rRNA기인Ⅴ구편단,제1례미발현기인돌변외,기여17례균발현2058화2059위점돌변,기중A2059G돌변10례,A2058G돌변5례,A2058T돌변2례.결론 23S rRNA기인Ⅴ구편단기인돌변가능여남경급주변지구Mg대대배내지류약물내약성상관.
Objective To detect point mutations associated with macrolide resistance in the 23S rRNA gene of M.genitalium.Methods Ninty-one patients with persistent or recurrent urethritis,who had been treated with macrolides in the past one month but achieved no obvious improvement,were included in this study.Urethral swab and first-void urine samples were collected from these patients.Gram's staining,culture and fluorescence-based quantitative PCR were carried out to detect Neisseria gonorrhoeae,Ureaplasma urealyticum and Chlamydia trachomatis respectively in these urethral swab samples,and PCR was performed to detect M.genitalium in the urine samples.For patients positive to only M.genitalium,nested PCR was conducted to amplify the domain Ⅴ of the 23S rRNA gene followed by direct automatic sequencing,and the resulting sequences were compared to the corresponding sequences of M.genitalium G37 strain submitted to the National Center for Biotechnology Information.Results Of the 91 patients,21 were positive for M.genitalium,18 for C.trachomatis,10 for U.urealyticum,2 for both M.genitalium and U.urealyticum,3 for both C.trachomatis and U.urealyticum,and 37 for none of the tested pathogens.The domain Ⅴ of the 23S rRNA gene,which was amplified from 18 of the 21 M.genitalium-positive samples,carried an A to G transition at position 2059 in 10 samples,an A to G transition at position 2058 in 5 samples,an A to T transition at position 2058 in 2 samples,but no point mutations in 1 sample.Conclusion The point mutations in domain Ⅴ of the 23S rRNA gene may contribute to macrolide resistance in M.genitalium in Nanjing and its surrounding areas.