实时荧光定量PCR结合探针检测MP的临床价值及MP耐药情况分析  

Role of real-time fluorescence quantitative PCR-binding probe in the determination of MP and MP drug resistance

作　　者：杜金龙 冯燕 李恒涛 DU Jinlong;FENG Yan;LI Hengtao(Department of Clinical Laboratory,Fengcheng Hospital of Shanghai Fengxian District,Shanghai 201411,China;Department of Pediatrics,Fengcheng Hospital of Shanghai Fengxian District,Shanghai 201411,China)

机构地区：[1]上海市奉贤区奉城医院检验科,上海201411 [2]上海市奉贤区奉城医院儿科,上海201411

出　　处：《检验医学》2020年第6期535-539,共5页Laboratory Medicine

摘　　要：目的评价实时荧光定量聚合酶链反应(PCR)结合探针检测对肺炎支原体(MP)的诊断价值,并分析MP的耐药突变情况,指导MP感染患儿的临床用药。方法选取2017年9月-2019年8月奉城医院收治并经诊断确诊的70例MP感染患儿及30例非MP感染患儿,对所有患儿咽拭子标本进行MP-DNA扩增,采用荧光探针法对MP与大环内酯类抗菌药物耐药突变率进行检测。收集所有患儿临床资料,并分析其与MP-DNA阳性检出及MP耐药突变的相关性。结果实时荧光定量PCR结合探针检测MP的敏感性和特异性分别为80.0%(56/70)和96.7%(29/30);双份血清MP-IgM抗体滴度呈4倍或4倍以上(升高或降低)变化组MP-DNA阳性检出率明显较单份血清滴度≥1∶160组高;患儿性别、年龄、入院时病程及入院前大环内酯类抗菌药物治疗史对于MP-DNA阳性的检出率均无显著影响;MP-DNA耐药突变率高达85.7%,入院前规律完成大环内酯类抗菌药物1个疗程组DNA耐药突变率为90.5%,明显高于未完成1个疗程组(80.0%)和未服用组(50.0%)。结论实时荧光定量PCR结合探针可作为检测MP的一种有效方法,为临床对MP感染患儿抗菌药物的使用提供依据,具有较高的临床应用价值。Objective To evaluate the role of real-time fluorescence quantitative polymerase chain reaction(PCR)-binding probe in the determination of Mycoplasma pneumoniae(MP),to analyze the drug resistance mutation of MP,and to guide the clinical medication for children with MP infection.Methods A total of 70 patients with MP infection and 30 patients without MP infection in Fengcheng Hospital of Shanghai Fengxian District from September 2017 to August 2019 were enrolled,and throat swab specimens were collected from children.MP-DNA amplification and fluorescence probe method were used to determine the drug resistance mutation of MP and macrolide antibiotics.The data of children were collected,and the correlation with MP-DNA positive and drug resistance mutation was analyzed.Results The sensitivity and specificity of PCR-binding probe for determining MP were 80.0%(56/70)and 96.7%(29/30),respectively.The MP-DNA positive determination rate of double serum MP-IgM antibody titer 4 times/4 times higher or lower group was higher than that of single serum titer≥1∶160 group.Sex,age,disease course and history of macrolide antibiotic treatment before admission had no effect on the positive rate of MP-DNA.The MP-DNA drug resistance mutation rate was 85.7%,and the rate of DNA drug resistance mutation in one course of treatment with macrolide antibiotics before admission was 90.5%,which was higher than those of untreated group(80.0%)and the group without finishing one course of treatment(50.0%).Conclusions Realtime fluorescence quantitative PCR-binding probe can be used as an effective method for MP determination,providing a reference for the clinical use of antibiotics in MP infection children,which has a high clinical application value.

关 键 词：肺炎支原体 儿童 实时荧光定量聚合酶链反应结合探针 耐药基因突变 

分 类 号：R446.1[医药卫生—诊断学]