3种方法检测儿童肺炎支原体感染的临床评估  

Clinical evaluation of 3 methods for the determination of Mycoplasma pneumoniae infection in children

作　　者：冯志山[1] 李贵霞[1] 刘建华[1] 郭巍巍[1] 张文超[1] 张丽霞[1] 黄慧[1] 王乐[1] FENG Zhishan;LI Guixia;LIU Jianhua;GUO Weiwei;ZHANG Wenchao;ZHANG Lixia;HUANG Hui;WANG Le(Institute of Pediatric Research,Children's Hospital of Hebei Province,Shijiazhuang 050031,Hebei,China)

机构地区：[1]河北省儿童医院儿科研究所

出　　处：《检验医学》2019年第9期808-811,共4页Laboratory Medicine

基　　金：河北省医学科学研究重点课题计划(20170402、20180616)

摘　　要：目的比较3种检测肺炎支原体(MP)感染的方法[被动凝集法、多重聚合酶链反应(PCR)及荧光定量PCR]的差异,探讨不同方法在不同年龄段及不同病程肺炎患儿中的应用价值。方法选取接受纤维支气管镜灌洗治疗的MP肺炎患儿213例。采用被动凝集法、多重PCR及荧光定量PCR分别检测血清MP抗体、痰液MP DNA及肺泡灌洗液(BALF)中的MP DNA。以荧光定量PCR在BALF中检出MP DNA为病原学判断标准,比较3种方法的一致性。将所有患儿按年龄分为<1岁组(8例)、1~3岁组(29例)、4~6岁组(97例)、>6岁组(79例),按病程分为≤7 d组和>7 d组。比较各组3种方法的阳性检出率。结果213例MP肺炎患儿中,被动凝集法、多重PCR及荧光定量PCR的阳性检出率分别为98.12%、76.06%和89.20%。以荧光定量PCR在BALF中检出MP DNA为病原学判断标准,多重PCR与荧光定量PCR的Kappa值为0.301,优于被动凝集法与荧光定量PCR的Kappa值(0.043)。按年龄分组,3种方法的阳性检出率均与年龄有关(P<0.05)。病程>7 d组荧光定量PCR阳性率高于≤7 d组(P=0.003),而被动凝集法和多重PCR的阳性检出率2个组之间差异无统计学意义(P>0.05)。结论在使用纤维支气管镜灌洗治疗的MP肺炎患儿中,荧光定量PCR与多重PCR的一致性优于被动凝集法,荧光定量PCR的阳性检出率与患儿病程相关。Objective To compare the differences among 3 methods[passive agglutination,multiplex polymerase chain reaction(PCR)and real-time quantitation PCR]for the determination of Mycoplasma pneumoniae(MP)infection,and to investigate their roles in different ages and disease stages of children with pneumonia.Methods A total of 213 children with MP pneumonia treated with fiberoptic bronchoscopy were enrolled.The passive agglutination,multiplex PCR and real-time quantitation PCR were used to determine serum MP antibodies,MP DNA in sputum and MP DNA in the bronchoalveolar lavage fluid(BALF),respectively.The results of realtime quantitation PCR for MP DNA in BALF were regarded as gold standard.The consistency of the 3 methods was compared.All the children were classified into<1-year-old group(8 cases),1-3-year-old group(29 cases),4-6-year-old group(97 cases)and>6-year-old group(79 cases).According to disease courses,they were classified into≤7 d and>7 d groups.The positive determination rates of the 3 methods were compared.Results Among the 213 children with MP pneumonia,the positive determination rates of passive agglutination,multiplex PCR and realtime quantitation PCR were 98.12%,76.06%and 89.20%,respectively.The results of real-time quantitation PCR for MP DNA in BALF were regarded as gold standard.The Kappa value of multiplex PCR with real-time quantitation PCR(0.301)was higher than that of passive agglutination with real-time quantitation PCR(0.043).The positive determination rates of the 3 methods were related to ages(P<0.05).The positive determination rate of real-time quantitation PCR in >7 d group was higher than that in ≤7 d group(P=0.003),while the positive determinationrates of passive agglutination and multiplex PCR had no statistical significance(P>0.05). Conclusions Among MPpneumonia patients treated with fiberoptic bronchoscopy,the consistency of real-time quantitation PCR and multiplexPCR is superior to passive agglutination,and the positive determination rate of real-time quantitation PCR is relatedwith

关 键 词：肺炎支原体 多重聚合酶链反应 荧光定量聚合酶链反应 肺泡灌洗液 

分 类 号：R446.1[医药卫生—诊断学]