肺炎支原体聚合酶链反应结合探针检测在儿童肺炎支原体肺炎诊断及耐药中的应用价值

摘要

Objective To evaluate the value of polymerase chain reaction (PCR) combined with probe detection method in diagnosis of Mycoplasma pneumonia (MP) pneumonia (MPP) in children and to analyze the factors influencing the diagnostic accuracy,and to identify the rate of MP mutation for drug resistance and the involving factors.Methods Two hundred and twenty-five children with MPP hospitalized in the Department of Respiratory Medicine,Beijing Children's Hospital,Capital Medical University between June 2015 and March 2016 were enrolled in this study.Nasopharyngeal swab samples from the participants within 24 hours of admission were detected by using PCR combined with fluorescence probes for MP-DNA and macrolide-resistant mutations.The information of age,sex,clinical symptoms,course of disease,duration by admission,the history of macrolide treatment and the increase or decrease of quadruple or more serum MP antibody titer were extracted from medical records within 4 weeks of treatment,which received further correlation analysis with the detection rate of MP-DNA and the drug resistance mutation.Results The sensitivity of the MPP by using the method of PCR combined with fluorescence probes was 80.4％ (181/225 cases),while the specificity was 98.0％ (99/101 cases).The MP-DNA positive rate for patients with double MP antibody 4 times increased during treatment was 88.8％ (71/80 cases),which was significantly higher than that of patients with antibody titer ≥1 ∶ 160 [75.9％ (110/145 cases)],and the difference was sigmficant(x =5.443,P =0.020).The positive rate of MP-DNA of patients had no obvious association with gender,age,and disease duration and macrolide treatment history before admission.Macrolide-resistant mutation rate of MP-DNA was 85.1％ (154/181 cases),macrolide-resistant mutation rate of MP for patients finishing one course of macrolide treatment when admission(89.6％)was higher than that of the patients without using macrolide and the patients treated with macrolide but not finishing one course of treatment (71.9％ and 86.6％),and the significant difference among the three groups was observed(x2 =4.454,P =0.035).Conclusions PCR combined with fluorescence probe for MP-DNA detection has a high accuracy for the diagnosis of MPP,and the overall mutation rate is high,suggesting that the clinical treatment of MPP needs to be adjusted according to drug resistance in children.%目的 评价肺炎支原体(MP)聚合酶链反应(PCR)结合探针检测方法对儿童MP肺炎(MPP)的诊断价值,并分析影响诊断准确性的因素,同时明确MP的耐药突变情况及影响因素.方法 选择2015年6月至2016年3月在首都医科大学附属北京儿童医院呼吸科住院并临床诊断为MPP的225例患儿及非支原体肺炎的101例患儿,对患儿入院24h内咽拭子标本进行MP-DNA扩增,荧光探针方法检测MP和大环内酯类抗生素耐药突变情况.将年龄、性别、临床症状、入院时病程、入院前大环内酯类抗生素治疗史及肺炎治疗4周内是否血清MP-抗体滴度4倍或4倍以上升高或减低等病历信息与MP-DNA检出及其耐药突变情况进行相关性分析.结果 PCR结合荧光探针方法检测MP肺炎的敏感性为80.4％(181/225例),特异性为98.0％ (99/101例).双份血清MP-抗体滴度4倍或4倍以上升高或减低组的MP-DNA检出率显著高于单次血清滴度≥1∶160组[88.8％(71/80例)比75.9％(110/145例)x2=5.443,P=0.020].性别、年龄、入院时病程长短及院外大环内酯类抗生素治疗与MP-DNA的检出率均未见相关性.MP-DNA耐药突变率为85.1％(154/181例),入院前已经规律使用大环内酯类抗生素治疗1个疗程组的DNA耐药突变率(89.6％)高于未使用组和使用但未完成1个疗程组的MP-DNA耐药突变率(71.9％及86.6％),3组比较差异有统计学意义(x2=4.454,P=0.035).结论 PCR结合荧光探针检测方法用于MPP诊断具有较高的敏感性和特异性,MP-DNA耐药突变率整体较高,儿童MPP的临床用药需要根据耐药性进行调整.