于静波,薛文成,张明磊,孟冬娅
YU Jing-bo,XUE Wen-cheng,ZHANG Ming-lei,MENG Dong-ya(Clinical Laboratory Department

• 1:沈阳军区总医院检验科

摘要(Abstract)：

目的监测我院2008～2011年人型支原体(Mh)对喹诺酮类药物的耐药性,并从作用靶位氨基酸残基变异这一角度研究其耐药机制。方法收集我院经液体药敏一体法培养的单独Mh感染标本,分别对编码MhⅡ型拓扑异构酶GyrA、GyrB、ParC、ParE亚基的相应基因进行PCR-测序分析,将DNA序列与GenBank中Mh模式菌株ATCC23114翻译后氨基酸序列比对分析(blastn)。按照药敏表型与氨基酸残基变异间的规律对Mh进行分组,并计算各组热点氨基酸残基变异的变异率,用四格表确切概率法进行统计学分析。结果对司帕沙星(SPA)耐药组(R)28株Mh均检出GyrA丝氨酸残基(Ser)83→亮氨酸残基(Leu),变异率为100.0%,高于非耐药组(S+I),差异有统计学意义(P<0.05)。对氧氟沙星(OFX)和左氧氟沙星(LEV)非敏感组(R+I)的40株Mh有36株检测出ParC134位赖氨酸残基(Lys)→精氨酸残基(Arg),变异率为90.0%,高于敏感组(S),差异有统计学意义(P<0.05);而对OFX和LEV R+I组有16株检出ParE 426位天冬氨酸残基(Lys)→天冬酰胺残基(Arg),变异率为40.0%,高于敏感组(S),但差异无统计学意义(P>0.05)。结论 Mh GyrA亚基Ser83→Leu是其对SPA耐药的原因之一;ParC亚基Lys134→Arg是其对OFX和LEV耐药的原因之一。
Objective To monitor the susceptibility of mycoplasma hominis to quinolones from 2008 to 2011 in our hospital,detect mutation of DNA Gyrase and topoisomerase Ⅳ in clinical isolates,and investigate the relation between mutation of DNA Gyrase and topoisomerase Ⅳ and the phenotypes to quinolones.Methods Clinical isolates were obtained from the urogenital specimens by liquid culture.PCR-sequencing analysised to the genes that respectivly code mycoplasma hominis type II topoisomerase GyrA,GyrB,ParC,ParE subunit.The sequences of GyrA,GyrB,ParC and ParE were compared to standard strain ATCC23114.Isolates was grouped according to susceptibility to quinolones,accouted their rate of mutation,and statistical analysis by fisher exact propability.All of 28 strains are resistant to sparfloxacin were detected GyrA serine83→leucine,mutation rate is 100.0%,higher than the nonresistance group(P<0.01).For 40 mycoplasma hominis isolates unsusceptible to ofloxacin and levofloxacin,the rate of lysine134→arginine mutation in ParC was 90.0%,higher than the sensitive group.The rate of aspartate(D)426→asparagine(N) mutation in ParE was 40.0%,but there was not significant difference(P>0.05) compared with the sensitive group positive rate.Results DNA gyrase serine83→leucine in GyrA is probably related with clinical isolates of mycoplasma hominis resistance to sparfloxacin;topoisomerase Ⅳ lysine134→arginine in ParC is probably related with clinical isolates of mycoplasma hominis resistance to ofloxacin and levofloxacin.Conclusions DNA gyrase serine83→leucine in GyrA is probably related with clinical isolates of mycoplasma hominis resistance to sparfloxacin;topoisomerase Ⅳ lysine134→arginine in ParC is probably related with clinical isolates of mycoplasma hominis resistance to ofloxacin and levofloxacin.

关键词(KeyWords)： 人型支原体;喹诺酮;DNA旋转酶;拓扑异构酶Ⅳ
mycoplasma hominis;quinolones;DNA gyrase;topoisomerase Ⅳ

Abstract:

Keywords:

基金项目(Foundation):

作者(Author): 于静波,薛文成,张明磊,孟冬娅
YU Jing-bo,XUE Wen-cheng,ZHANG Ming-lei,MENG Dong-ya(Clinical Laboratory Department

DOI: 10.14066/j.cnki.cn21-1349/r.2012.10.008

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