化学发光免疫分析法快速检测血清登革热病毒NS1抗原方法的建立及初步应用评价  

Establishment and Preliminary Evaluation of Chemiluminescence Immunoassay Method for Rapid Detection of Dengue Virus NS1 Antigen in Serum

作　　者：陈翠翠 梁焕坤 钟树海 陆嫣红 李来庆 CHEN Cui-cui;LIANG Huan-kun;ZHONG Shu-hai;LU Yan-hong;LI Lai-qing(Guangzhou Youdi Biotechnology Co.Ltd,Guangzhou 510663,China;Jinan Laide Biotechnology Co.Ltd,Jinan 271100,China)

机构地区：[1]广州优迪生物科技股份有限公司,广州510663 [2]济南市来德生物科技有限公司,济南271100

出　　处：《现代检验医学杂志》2023年第3期176-179,194,共5页Journal of Modern Laboratory Medicine

摘　　要：目的建立一种化学发光免疫分析法(chemiluminescence immunoassay,CLIA)用于血清中登革热病毒(dengue virus,DENV)NS1抗原的定量检测。方法先制备NS1单抗与吖啶酯发光剂的偶联物,同时活化磁珠并标记抗NS1的配对单抗,基于双抗体夹心检测方法的技术原理构建定量检测血清中NS1抗原浓度的化学发光检测方法。通过对其灵敏度、特异度和参考区间等试验评估其检测性能。取85例临床确诊的登革热阳性血清样本和20份健康志愿者阴性血清样本,采用美国Cortez公司的登革热NS1快速检测试剂盒和该方法进行临床样本验证。结果该方法对血清中NS1抗原的检测灵敏度为1.12ng/ml,线性范围在0.1~1000ng/ml;与血清中常见干扰物质的交叉反应率均低于2%,检测特异度较好;参考区间的cut off值为3.66ng/ml;对临床样本的检测准确度较高,与Cortez公司NS1检测试剂盒比较无显著性差异(χ^(2)检验P=1.000,一致性检验,Kappa=0.876)。结论建立了一种可定量检测血清中登革热病毒NS1抗原的CLIA法,该方法具有高灵敏度、高准确度、操作简单和方便快捷等优点,有望为登革热临床样品的早期准确筛查提供一种新的选择。Objective To establish a chemiluminescence immunoassay(CLIA)for quantitative detection of dengue virus(DENV)NS1 antigen in serum.Methods The conjugation of NS1 monoclonal antibody and acridine ester luminescent agent was prepared,and the magnetic beads were activated at the same time to label the paired monoclonal antibody against NS1.Based on the technical principle of double antibody sandwich detection method,a CLIA method was constructed to quantitatively detect the concentration of NS1 antigen in serum.The detection performance was evaluated by sensitivity assays,specificity assays and reference interval assays.85 dengue positive serum samples and 20 negative serum samples of healthy volunteers were taken for clinical sample verification with the dengue NS1 rapid detection kit of Cortez and this method.Results The sensitivity of this method to detect NS1 antigen in serum was 1.12 ng/ml,and the linear range was 0.1~1000 ng/ml.The cross reaction rates with common interfering substances in serum were all lower than 2%,and the detection specificity was good.The cut off value of the reference interval was 3.66 ng/ml.The detection accuracy of clinical samples was high,and there was no significant difference compared with Cortez NS1 detection kit(χ^(2) test,P=1.000 and Kappa=0.876).Conclusion A CLIA method for quantifying dengue virus NS1 antigen in serum has been established,which has the advantages of high sensitivity and accuracy,simple operation,convenience and rapidity,and it is expected to provide a new choice for early and accurate screening of dengue clinical samples.

关 键 词：登革热病毒 NS1抗原 化学发光免疫分析法 吖啶酯 磁珠 

分 类 号：R373.33[医药卫生—病原生物学] R446[医药卫生—基础医学]