PCR-毛细电泳片段分析法检测毛细支气管炎病原的临床价值探讨

PCR-capillary electrophoresis fragment analysis in detection of pathogens for children with bronchiolitis

目的探讨PCR-毛细电泳片段分析法(PCR-CEFA)检测毛细支气管炎病原的临床应用价值。方法选取2018年1至12月温州医科大学附属第二医院、育英儿童医院儿科收治的因毛细支气管炎住院的≤24个月且喘息次数≤2次的患儿932例,采集患儿鼻咽分泌物,采用PCR-CEFA检测呼吸道合胞病毒(RSV)、鼻病毒(RV)、流感病毒(Flu)、副流感病毒(HPIV)、腺病毒(ADV)等8种病毒核酸;直接免疫荧光法(DFA)检测RSV、HPIV、Flu、ADV抗原;并收集患儿性别、年龄、既往喘息史、湿疹史、哮喘家族史、家族过敏性疾病史、总IgE、血嗜酸性粒细胞计数(B-Eos)等资料。比较两种方法、不同年龄组患儿病毒检出率及RSV组与RV组患儿流行病学资料及实验室检查结果。结果PCR-CEFA病毒检出率为88.2%,高于DFA的33.6%,差异有统计学意义(P<0.01)。PCR-CEFA对RSV、HPIV、Flu、ADV的检出率均高于DFA,差异均有统计学意义(均P<0.01)。以DFA为参照,PCR-CEFA检测RSV、HPIV、Flu、ADV的特异度均达80%以上。≤12月龄患儿RSV检出率高于>12~24月龄患儿,RV、ADV、HBoV检出率均低于>12~24月龄患儿,差异均有统计学意义(均P<0.05)。RV组患儿年龄、B-Eos升高比例均明显高于RSV组,差异均有统计学意义(均P<0.01)。结论与DFA比较,PCR-CEFA病毒检出率更高,病毒检测种类更多,具有良好的特异度,有利于早期明确毛细支气管炎病原。

Objective To apply PCR-capillary electrophoresis fragment analysis(PCR-CEFA)in the pathogen detection of bronchiolitis.Methods A total of 932 children aged≤24 months who were clinically diagnosed as bronchiolitis in the Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University From January to December 2018,were enrolled in the study.The clinical data were collected,including gender,age,history of wheezing and eczema,family history of asthma and allergic diseases,total IgE,blood eosinophil count(B-Eos).The nasopharyngeal secretion samples were collected,PCR-CEFA was used to detect 8 viral nucleic acids,including respiratory syncytial virus(RSV),rhinovirus(RV),influenza virus(Flu),human parainfluenza virus(HPIV),adenovirus(ADV),and human boca virus(HBoV),the direct immunofluorescence assay(DFA)was used to detect the antigen of RSV,HPIV,Flu,ADV.The detection rates of pathogens between two methods,and among children of different age groups were compared,the epidemiological data and laboratory test results of the RSV group and the RV group were also compared.Results The virus detection rates of PCR-CEFA was 88.2%,which was higher than that of DFA(33.6%,P<0.01).The detection rate of PCR-CEFA for RSV,HPIV,Flu and ADV was higher than that of DFA(all P<0.01).Using DFA as a reference,the specificity of PCR-CEFA to detect RSV,HPIV,Flu,ADV was more than 80%.The detection rate of RSV in children aged≤12 months was higher than that of children aged>12 months,while the detection rates of RV,ADV,and HBoV in children aged≤12 months were lower than those of children aged>12 months(all P<0.05).The age and the proportion of B-Eos increase in the RV group were significantly higher than those in the RSV group(both P<0.01).Conclusion Compared with DFA,PCR-CEFA has a higher virus detection rate,more types of virus detected,and good specificity,which is conducive to early identification of the pathogens in children with bronchiolitis.