支原体DNA载量和耐药基因23SrRNA的2063和2064位点突变检测在儿童支原体肺炎诊疗的应用

Application of Mycoplasma DNA Load and 2063 and 2064 Point Mutation Detection of Drug Resistance Gene 23SrRNA in Diagnosis and Treatment of Mycoplasma Pneumonia in Children

目的了解支原体DNA载量和耐药基因23SrRNA的2063和2064位点突变检测在肺炎支原体肺炎(MPP)的早期诊断和大环内酯类药物耐药应用情况分析。方法收集2017年3月至2019年9月昆明医科大学第一附属医院儿科因下呼吸道感染住院患儿224例为研究对象,其中男122例,女102例。年龄1~13岁,平均(5.57±3.34)岁,入院24 h严格规范采集痰液和咽拭子的标本并及时送检。采用PCR法检测MP-DNA阳性率及耐药基因23SrRNA的2063和2064位点突变情况,并对临床资料进行统计分析。结果224例下呼吸道感染患儿中,MP感染71例,感染率为31.69%,不同年龄组间MP感染率比较差异有统计学意义(P<0.05),且感染率随着年龄的增长而升高;不同性别患儿MP阳性率比较,差异无统计学意义(P>0.05)。其中检出MP-DNA阳性27人,阳性率:38%,其中检出23SrRNA的2063和2064位点突变耐药基因19人。灵敏度为70.4%,特异度为100%,结论采用PCR法对呼吸道分泌物MP-DNA载量的检测在儿童MPP早期诊断中因特异性和敏感性高、简便易行、儿童依从性好等特点得到推广,同时检测呼吸道分泌物MP耐药基因23SrRNA的2063和2064位点突变检测及时指导治疗值得肯定。

Objective To understand the detection of Mycoplasma DNA load and resistance mutations of23 SrRNA at 2063 and 2064 sites in the early diagnosis of Mycoplasma pneumoniae pneumonia(MPP)and analysis of the application of macrolide resistance.Methods From March 2017 to September 2019,224 children hospitalized for lower respiratory infections in the Department of Pediatrics of the First Affiliated Hospital of Kunming Medical University were collected as subjects,including 122 males and 102 females.The age ranged from 1 to 13 years and the average age was(5.57±3.34)years.Within 24 hours of admission,samples of sputum and throat swabs were strictly collected and submitted for inspection in time.The PCR method was used to detect the positive rate of MP-DNA and the 2063 and 2064 mutations of the 23 SrRNA resistance gene,and the clinical data were analyzed statistically.Results Among the 224 children with lower respiratory tract infections,71 were MP infected,and the infection rate was 31.69%.The difference in MP infection rate between different age groups was statistically significant(P<0.05),and the infection rate increased with age.There was no significant difference in MP positive rate among children of different genders(P>0.05).Among them,27 were positive for MP-DNA,and the positive rate was 38%.Among them,19 were detected resistance genes for 2063 and 2064 mutations in 23 SrRNA.The sensitivity is 70.4%and the specificity is 100%.Conclusions The detection of MP-DNA load in respiratory secretions using PCR method in early diagnosis of MPP in children has high specificity and sensitivity,is easy to implement,and has good compliance for children.The simultaneous detection of 2063 and 2064 loci at the same time for the detection of MP resistance gene 23 SrRNA in respiratory secretions is worthy of timely guidance and treatment.