呼吸道病原体多重核酸扩增在老年患者呼吸系统感染检测中的应用  

Application of multiple RT-PCR assay for respiratory system infection in elderly patients

作　　者：宋国威[1] 潘婧[1] 梁芸[1] 马学军[2] 万玲玲[1] 李玉雪[1] 李立新[1] Song Guowei;Pan Jing;Liang Yun;Ma Xuejun;Wan Lingling;Li Yuxue;Li Lixin(The First Hospital of Shijiazhuang,Shijiazhuang 050000,China;Key Laboratory for Medical Virology,National Health and Family Planning Commission,National Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China)

机构地区：[1]石家庄市第一医院检验科,050000 [2]中国疾病预防控制中心病毒病预防控制所国家卫生健康委员会医学病毒和病毒病重点实验室,北京102206

出　　处：《中华实验和临床病毒学杂志》2020年第3期299-303,共5页Chinese Journal of Experimental and Clinical Virology

基　　金：石家庄市科技技术研究与发展指导计划项目(171462093)。

摘　　要：目的了解多重RT-PCR方法检测13种常见呼吸道感染病原体,在老年患者呼吸系统感染病原体检测中的应用价值。方法317例2016年6月至2017年5月在石家庄市第一医院老年科就诊的具有呼吸系统感染症状的老年患者,年龄60~98岁,采集其抽吸痰液,提取病毒DNA/RNA,利用多重RT-PCR技术扩增,通过毛细管电泳,同时检测13种呼吸道病原体;并以直接测序方法和荧光实时PCR法为标准,评价该试剂的临床性能。结果使用直接测序和荧光实时PCR结果为参照,多重RT-PCR技术的准确性为99.93%(95%CI:99.79%~99.98%),阳性预测值为99.24%(95%CI:97.78%~99.74%),阴性预测值为100%(95%CI:99.9%~100%),Cohen’skappa值为0.9958(95%CI:0.9652~1.026)。在317例老年患者中,呼吸道病原阳性检出率为75.7%(240/317),其中甲型流感病毒最高,为20.2%(64/317),主要为H3亚型(70.3%(45/64);其次为鼻病毒、腺病毒、肺炎支原体和乙型流感病毒,检出率分别为15.5%(49/317)、13.6%(43/317)、11.0%(35/317)和10.1%(32/317)。此外,同时检出2种以及2种以上病原体的老年患者为81例,检出率为25.6%(81/317)。结论多重RT-PCR方法能满足临床对呼吸道病原体检测的需求,并为老年患者呼吸系统感染的防治提供有效的临床资料。Objective To understand application value of pathogen detection in elderly patients with respiratory infection by using multiple RT-PCR to detect 13 common pathogens of respiratory tract infection.Methods This is a retrospective study,which included 317 elderly patients,aged 60 to 98 years with respiratory symptoms,who visited Geriatrics of the First Hospital of Shijiazhuang between June 2016 and May 2017.Sputum of each patient was collected.After the viral DNA/RNA was extracted,13 respiratory pathogens were detected by multiplex RT-PCR and capillary electrophoresis.The clinical performance of the assay was evaluated in comparison to direct sequencing and Real-time PCR.Results Compared with the direct sequencing and Real-time PCR,the accuracy of multiple RT-PCR was 99.93%(95%CI:99.79%-99.98%),positive predictive value was 99.24%(95%CI:97.78%-99.74%),negative predictive value was 100%(95%CI:99.9%-100%),and Cohen’s kappa was 0.9958(95%CI:0.9652-1.026).Among the 317 elderly patients,the positive rate of respiratory infection was 75.7%(240/317).The highest was influenza A,20.2%(64/317),mainly H3N2,70.3%(45/64),followed by rhinovirus,adenovirus,Mycoplasma pneumoniae and influenza B,with the positive rates 15.5%(49/317),13.6%(43/317),11.0%(35/317)and 10.1%(32/317),respectively.In addition,81(25.6%)elderly patients were infected with two or more pathogens.Conclusions Multiple RT-PCR meet the needs of clinical detection of respiratory pathogens and provide information for the prevention and treatment of respiratory system Infection.

关 键 词：呼吸道感染 毛细管电泳 老年患者 逆转录聚合酶链反应 

分 类 号：R440[医药卫生—诊断学] R56[医药卫生—临床医学]