2019年1月至2023年4月河南省南阳市甲型流感病毒流行变迁及实验室检测方法学差异分析

Epidemic changes of influenza A virus and analysis of differences in laboratory detection methods in Nanyang,Henan from September 2019 to April 2023

目的探讨河南省南阳市甲型流感病毒(IAVs)的流行变迁及多中心实验室不同检测方法学检测IAVs的差异分析。方法收集河南省南阳市中心医院、南阳市第一人民医院、南阳康圣医学检验所和郑州金域临床检验中心有限公司2019年1月至2023年4月期间,经临床医生诊疗疑似IAVs感染发热患者外周静脉血、咽拭子和肺泡灌洗液等标本,采用间接免疫荧光法(IFA)、磁微粒化学发光法(CLIA)、胶体金法、一代测序法、反转录-聚合酶链式反应(RT-PCR)及靶向高通量测序技术(tNGS)分别进行检测,分析各中心实验室不同检测方法学优劣及关联,以及分析IAVs流行变迁。结果发热门诊送检IAVs标本检测量较大,IAVs患者分布于各年龄段,各人群均易感。2020—2022年期间IAVs处于低流行状态,2019年和2023年至今,IAVs出现大流行。tNGS、一代测序与RT-PCR检测IAV-RNA阳性率比较,三者之间差异无统计学意义(χ^(2)=1.449、0.838,P=0.229、0.360),IAV-IgM(欧蒙)对IAVs检出阳性率与上述方法学之间差异无统计学意义(χ^(2)=2.447,P=0.118),IAV-IgM(西班牙)、IAV-IgM(CLIA)对IAVs检出阳性率较低。tNGS与一代测序可以对IAVs进行分型,检测IAVs及亚型IAV-H3N2阳性率之间差异具有统计学意义(u=4.335、4.213,P<0.001)。当IAV-RNA的Ct值<25时,患者咽拭子中IAV-Ag敏感度高达88/90=97.78%,当Ct值≥30时,IAV-Ag阳性检出率仍保持在40.00%以上,金标法检测IAV-Ag敏感度可达77.22%。一代测序和tNGS显示出检测IAV-RNA及其亚型的优势,IFA和CLIA显示出检测肺炎支原体IgM的优势。结论河南省南阳市IAVs常年流行,冬季为流行季。不同中心实验室不同检测方法对IAVs阳性检出率不同,选择IAVRNA联合IAV-IgM检测可为临床提供准确的IAVs检测结果。

Objective To understand the epidemiological characteristics and method specific detection of Influenza A viruses in Nanyang,Henan province.Methods Peripheral venous blood,pharyngeal swabs and alveolar lavage fluid samples of fever patients with suspected influenza A virus infection were collected from Nanyang Central Hospital,Nanyang First People's Hospital,Nanyang Kangsheng Medical Laboratory Corporation and Zhengzhou Jinyu Clinical Testing Center Corporation from September 2019 to April 2023.Indirect immunofluorescence(IFA),magnetic particle chemiluminescence(CLIA),colloidal gold method,first generation sequencing,reverse transcription-polymerase chain reaction(RT-PCR)and Target next generation sequencin(tNGS)were used for the sample detection.The advantages,disadvantages,and correlations of different detection methods were analyzed,and then the spread of influenza A virus was analyzed.Results A large number of the samples were sent from fever clinics,the influenza A virus infection cases were distributed in all age groups,indicating that all populations were susceptible to influenza A virus,The spread level of influenza A virus was low during 2020-2022,but was high in 2019 and 2023.There were no significant differences in the positive rate of RNA of influenza A virus detected by tNGS,first-generation sequencing,and RT-PCR(χ^(2)=1.449,0.838,P=0.229,0.360),including IAV-IgM detection(EUROIMMUN).tNGS and the first-generation sequencing can be used to classify influenza A virus,but the differences in detecting influenza A virus and influenza A(H3N2)virus were significant(u=4.335,4.213,P<0.001).When the Ct value of RNA of influenza A virus was less than 25,the detection rate of glAV-A in the patient's throat swabs was relatively stable,with a sensitivity of 97.78%,when the Ct value was≥30,the detection rate of IAV-Ag remained above 40.00%,and the total sensitivity of the gold standard method for detecting IAV-Ag could reach 77.22%.The first generation sequencing and tNGS showed advantage in detecting RNA of influenza A virus,but iFA and CLIA showed advantage in detecting IgM of Mycoplasma pneumoniae Conclusion Influenza A virus spread all the year round in Nanyang,but mainly in winter.The detection rate of influenza A virus varied with different methods in different laboratories.The co-detection of RNA and IgM of influenza A virus could provide accurate influenza A virus detection results in clinical practices.