摘要
目的 比较甲型流感病毒抗原即时检验(POCT)免疫荧光法、乳胶免疫层析法和免疫胶体金法的临床诊断效能.方法 以qRT-PCR为诊断标准,比较3种甲型流感病毒抗原POCT对甲型流感病毒的检出限、分析特异性和在临床样本检测中的诊断灵敏度和诊断特异性.结果 免疫荧光法、乳胶免疫层析法和免疫胶体金法对H1N1pdm09 的检出限分别为 1∶10000、1∶1 000 和 1∶100,对季节性流感 H1N1、H1N1(PR8 株)、H3N2(Aichi 株)、禽流感病毒H6N2和H7N3的检出限分别为1∶100 000、1∶10 000和1∶1 000.检出限从低到高依次为免疫荧光法、乳胶免疫层析法和免疫胶体金法.3种甲型流感病毒抗原POCT对除甲流外的25种临床呼吸道感染常见病原体均无交叉反应.与qRT-PCR相比,免疫荧光法、乳胶免疫层析法、免疫胶体金法的诊断灵敏度分别为74.63％、56.72％和40.30％,免疫荧光法的诊断灵敏度高于乳胶免疫层析法,差异有统计学意义(x2=46.13,P＜0.05),免疫荧光法的诊断灵敏度高于免疫胶体金法,差异有统计学意义(x2=71.94,P＜0.05),乳胶免疫层析法的诊断灵敏度高于免疫胶体金法,差异有统计学意义(x2=103.54,P＜0.05).免疫荧光法、乳胶免疫层析法、免疫胶体金法的诊断特异性分别为96.41％、100.00％、100.00％,受试者工作特征(ROC)曲线下面积(AUC)分别为0.86、0.78和0.70.结论 免疫荧光法在3种甲型流感病毒抗原POCT中表现最佳,具有最低的检出限、最高的诊断灵敏度和AUC,以及较好的分析特异性和诊断特异性,可满足临床对甲型流感病毒抗原的快速检测需求.
Abstract
Objective To compare the clinical diagnostic performance of influenza A virus antigen point-of-care testing(POCT)by immunofluorescence assay,latex immunochromatographic assay,and immunochromatographic colloidal gold assay.Methods qRT-PCR was used as the diagnostic standard;the detection limit,analytical specificity,and diagnostic sensitivity and specificity of three influenza A virus antigen POCT methods were compared in clinical samples.Results The detection limits of immunofluorescence assay,latex immunochromatographic assay,and immunochromatographic colloidal gold assay for H1N1pdm09 were 1∶10 000,1∶1 000,and 1:100,respectively,and for seasonal influenza H1N1,H1N1(PR8 strain),H3N2(Aichi strain),avian influenza virus H6N2,and H7N3 were 1∶100 000,1∶10 000,and 1∶1 000,respectively.The detection limits were in the order of immunofluorescence assay,latex immunochromatographic assay,and immunochromatographic colloidal gold assay from low to high.None of the three influenza A virus antigen POCT methods showed cross-reaction with 25 common pathogens of clinical respiratory infections other than influenza.Compared with qRT-PCR,the diagnostic sensitivity of immunofluorescence assay,latex immunochromatographic assay,and immunochromatographic colloidal gold assay were 74.63％,56.72％,and 40.30％,respectively.The diagnostic sensitivity of immunofluorescence assay was significantly higher than that of latex immunochromatographic assay and immunochromatographic colloidal gold assay(x2=46.13,71.94;all P＜0.05),and the diagnostic sensitivity of latex immunochromatographic assay was significantly higher than that of immunochromatographic colloidal gold assay(x2=103.54,P＜0.05).The diagnostic specificity of immunofluorescence assay,latex immunochromatographic assay,and immunochromatographic colloidal gold assay were 96.41％,100.00％,and 100.00％,respectively.The area under the curve(AUC)of receiver operating characteristic curve(ROC)of immunofluorescence assay,latex immunochromatographic assay,and immunochromatographic colloidal gold assay were 0.86,0.78 and 0.70,respectively.Conclusions Immunofluorescence assay showed the best performance among the three influenza A virus antigen POCT methods,with the lowest detection limit,the highest diagnostic sensitivity and AUC,and good analytical specificity and diagnostic specificity.It can meet the clinical demand for rapid detection of influenza A virus antigen.
基金项目
广州市科技局基础研究计划(202102100003)
广东省本科高等学校教学质量与教学改革工程建设医学检验创新联合实验室项目(粤教高函[2021]29号)
NETL
NSTL
万方数据