多重荧光PCR技术在诊断儿童急性呼吸道感染中的应用

目的评估多重荧光PCR技术在检测儿童急性呼吸道感染病原体中的临床应用价值。方法随机选择我院急性呼吸系统感染患儿502例,采集鼻咽拭子,分别采用多重荧光PCR技术和胶体金、免疫荧光等方法检测呼吸道病原体,对不同检测结果采用二代测序方法进行验证,评估多重PCR技术在检测呼吸道病原体中的价值。结果502份标本采用多重PCR技术检测出10种病原体,共计200株,阳性标本118份(阳性率23.50%),混合感染率11.4%,与测序结果一致,与其它免疫方法比较,除甲乙流病毒检测一致性好(kappa>0.8),其它病原体检测的检测一致性较差(kappa<0.4)。结论多重荧光PCR技术具有灵敏、特异、准确、快速高效等优点,而胶体金法、免疫荧光法灵敏度低、结果判读主观性强。多重荧光PCR技术对于诊断儿童呼吸系统感染具有良好的临床应用价值。

Application of multiplex fluorescent PCR assay in the diagnosis of acute respiratory infections in children

Objective To investigate the clinical value of multiplex fluorescent PCR in the detection of common pathogens of acute respiratory tract infections in children.Methods Nasopharyngeal swabs from 502 children with acute respiratory infection in our hospital were randomly collected.The pathogens were detected by multiplex fluorescent PCR technology,colloidal gold,and immunofluorescence method respectively.The second-generation sequencing method was used to verify the different detection results,and the value of multiplex PCR in detecting respiratory pathogens was evaluated.Results A total of 200 strains of 10 pathogens were detected by multiple fluorescent PCR in 502 samples,of which 118 were positive(positive rate 23.50%),and the mixed infection rate was 11.4%,which was consistent with the sequencing results.Compared with other immunoassay methods,the multiple fluorescent PCR has good consistency in the detection of influenza A and B virus(Kappa>0.8),but the detection consistency of other pathogens is poor(Kappa<0.4).Conclusion The multiplex fluorescent PCR assay has the advantages of sensitivity,specificity,accuracy,fast and high efficiency,while the colloidal gold method and immunofluorescence method have low sensitivity and strong subjective interpretation of results.Multiple fluorescent PCR technology has good clinical application value for the diagnosis of childhood respiratory infections.