儿童肺炎支原体对大环内酯类药物耐药机制研究  

The research of Mycoplasma pneumoniae in children resistance to macrolides

作　　者：肖光文[1] 李舟文[1] 张国雄[2] 徐美兰[1] 乔亚峰 曾令斌[1] 

机构地区：[1]嘉应学院医学院,广东梅州514031 [2]梅州市人民医院,广东梅州514031

出　　处：《中国热带医学》2017年第11期1067-1071,共5页China Tropical Medicine

基　　金：广东省医学科学研究基金(No.B2015036);梅州市科技计划项目(No.2015B041)

摘　　要：目的研究临床分离的肺炎支原体(Mycoplasma pneumoniae,Mp)耐药菌株对大环内酯类(macrolide,ML)药物耐药的机制。方法收集经快速鉴定培养法鉴定的对红霉素耐药的Mp阳性培养物共计171例,经聚合酶链反应(polymerase chain reaction,PCR)法筛选并确定为Mp耐药菌株后,分别进行23S rRNA、L4和L22基因PCR扩增分析其耐药基因突变位点,同时行靶位修饰erm A/B/C基因和外排泵mefA、msrA/B基因的PCR扩增,分析Mp对ML耐药的可能机制。结果 165例Mp阳性培养物经PCR法确定为Mp临床耐药菌株;这些菌株里,全部菌株均存在23S rRNA耐药基因的碱基突变,74例(44.8%)存在核糖体蛋白L22碱基突变,37例(22.4%)存在核糖体蛋白L4碱基突变;其中,出现23S rRNA的Ⅴ区A2063G突变152例(92.1%),其次是Ⅱ区640位点A缺失39例(23.6%)、Ⅴ区2471位点C缺失26例(15.8%)、Ⅱ区G759A突变23例(13.9%);L4中发生C162A及A430G突变28例(17.0%),另外3例出现A268G突变、2例出现C428T突变;L22中发生498或499插入A 64例(38.8%),其次是T279C突变15例(9.1%);所有Mp临床耐药菌株均未扩增出erm A/B/C基因产物,2例扩增出mefA基因产物,1例扩增出msrA/B基因产物。结论该地区出现的Mp耐药机制以靶位突变为主,有些还存在外排泵机制,其携带的外排泵可能属于ATP结合区(ATP binding cassette,ABC)转运器。Objective In order to study on resistant mechanism against macrolides （ML） of Mycoplasma pneumoniae （Mp） resistant strains from clinical. Methods A total of 171 cases of erythromycin resistant Mp positive cultures were identified by rapid identification culture, Mp resistant strains were screened and identified by PCR method, after that 23S rRNA,L4 and L22 genes were amplified by PCR and their resistance gene mutation sites were analyzed. Meanwhile ermA/B/C gene about target modification , MefA and msrA/B genes about drug efflux pump were amplified by PCR, the possible mechanism of Mp resistance to ML was analyzed finally. Results A total of 165 Mp positive cultures were identified as Mp resistant strains by PCR; all of these strains, 165 strains were 23S rRNA gene mutation, 74 cases （44.8%） of ribosomal protein L22 gene mutation, 37 cases （22.4%） of ribosomal protein L4 gene mutation;among them, 152 cases （92.1%） of 23S rRNA V A2063G mutation, followed by 39 cases （23.6%） II 640 locus A deletion, 26 cases （15.8%） V 2471 locus C deletion, 23 cases （13.9%） appeared in the II area of the G759A/A mutation; in 28 cases （17.0%） the C162A and A4]0G mutation, the other 3 cases with A268G mutation, C428T mutation occurred in 2 cases; L22 64 cases （38.8%） occurred in 498 or 499 into A, followed by 15 cases （9.1%） had T279C mutation;ErmA/B/C gene products were not amplified in all clinical drug-resistant strains of Mp, and mefA gene products were amplified in 2 cases, and msrA/B gene products were amplified in 1 cases. Conclusion These results suggest that the Mp resistance in this region was mainly caused by target mutation, and some of them also had efflux pump mechanism, and the efflux pump they carry might belong to the ABC transporter.

关 键 词：肺炎支原体 儿童 大环内酯类 耐药性 

分 类 号：R375[医药卫生—病原生物学]