基于流式量子点微球技术的甲乙型流感病毒抗原检测方法的建立和初步应用分析  

Establishment and Preliminary Application Analysis of A Multiplex Detection Method for Influenza A and B Virus Antigen Based on Quantum Dot-encoded Microsphere Flow Cytometry Technology

作　　者：夏成静 李宝花 郭燕妳 周小合 张润玲 牛英波 XIA Chengjing;LI Baohua;GUO Yanni;ZHOU Xiaohe;ZHANG Runling;NIU Yingbo(Department of Clinical Laboratory,Shenzhen Guangming District People’s Hospital(West Wing),Guangdong Shenzhen 518106,China;Nanjing Vazyme Biotech Co.Ltd,Nanjing 210033,China)

机构地区：[1]深圳光明区人民医院(西)检验科,广东深圳518106 [2]南京诺唯赞医疗科技有限公司,南京210033

出　　处：《现代检验医学杂志》2024年第1期126-130,共5页Journal of Modern Laboratory Medicine

基　　金：深圳光明区软科学研究项目(项目批号2021R01097)。

摘　　要：目的建立基于流式量子点微球技术的甲型流感病毒(FluA)和乙型流感病毒(FluB)抗原联检方法,为常见呼吸道病毒抗原多重检测打下基础。方法分别使用自制的不同量子点编码微球和小鼠抗FluA/FluB核蛋白(NP)单克隆抗体进行偶联,在流式细胞仪上对已知浓度的甲乙型流感病毒抗原分别和同时进行检测,对检测条件进行优化,建立甲乙型流感病毒抗原联检方法,利用此方法对临床样本进行检测,与实时荧光定量PCR法(quantitative real-time PCR,qPCR)进行比对,验证此方法的临床性能。结果建立了甲乙型流感病毒抗原联检方法,该方法的甲乙型流感病毒抗原的检出限分别为26.1 pg/ml和10.7 pg/ml,测量范围均为15.3~250000 pg/ml。对临床样本检测,与qPCR比对一致性良好,阳性符合率为57.4%,阴性符合率为100%,总符合率71.6%,并优于临床常用胶体金试剂(阳性符合率为56.49%,阴性符合率为99.75%)。结论此流式量子点微球多重检测方法可以用于甲乙型流感病毒抗原的联检,其灵敏度较高、特异度好、检测范围广,可以为呼吸道常见病毒多重检测打下良好基础,在临床上具有应用前景。Objective To establish a multiplex assay method for the simultaneous detection of FluA and FluB virus(IBV)antigen based on the flow cytometry(FCM)quantum dot-encoded bead technologies,laying the foundation for the assay of multiple respiratory virus biomarkers.Methods Coupling was performed for FluA and FluB nucleoprotein(NP)monoclonal antibodies using self-made quantum dot-encoded beads,separately.FCM was used to detect known concentrations of FluA and FluB antigens separately and simultaneously,optimize the detection conditions,and establish a joint detection method for FluA and FluB antigens.Compared with the quantitative real-time PCR(qPCR)method,clinical samples were used to evaluate the clinical performance of this joint detection method.Results The joint detection method for FluA and FluB antigens was established,with detection limits of 26.1 pg/ml and 10.7 pg/ml,respectively,and measurement ranges of 15.3~250000 pg/ml.The joint detection method for clinical sample evaluation was well correlated with the qPCR,with a positive coincidence rate of 57.4%,a negative coincidence rate of 100%,and a total coincidence rate of 71.6%.In addition,the joint detection method was superior to colloidal gold immunochromatographic strip assay commonly used in clinical practice(positive coincidence rate of 56.49%,negative coincidence rate of 99.75%).Conclusion The FCM quantum dot-encoded bead multiplex assay can be used for the joint detection of FluA and FluB antigens,which have a high sensitivity,good specificity and wide detection range.It may lay a good foundation for the multiplex detection of common respiratory viruses,and has clinical application prospects.

关 键 词：甲型流感病毒 乙型流感病毒 量子点编码微球 液相蛋白芯片 

分 类 号：R373.13[医药卫生—病原生物学] R446.62[医药卫生—基础医学]