[1]夏成静,李宝花,郭燕妳,等.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[J].现代检验医学杂志,2024,39(01):126-130.[doi:10.3969/j.issn.1671-7414.2024.01.023]
 XIA Chengjing,LI Baohua,GUO Yanni,et al.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[J].Journal of Modern Laboratory Medicine,2024,39(01):126-130.[doi:10.3969/j.issn.1671-7414.2024.01.023]
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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()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第39卷
期数:
2024年01期
页码:
126-130
栏目:
研究简报·实验技术
出版日期:
2024-01-15

文章信息/Info

Title:
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
文章编号:
1671-7414(2024)01-126-05
作者:
夏成静1李宝花1郭燕妳1周小合1张润玲1牛英波2
[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]
Author(s):
XIA Chengjing1 LI Baohua1 GUO Yanni1 ZHOU Xiaohe1 ZHANG Runling1 NIU Yingbo2
[1. Department of Clinical Laboratory, Shenzhen Guangming District People’s Hospital (West Wing), Guangdong Shenzhen 518106, China;2. Nanjing Vazyme Biotech Co. Ltd, Nanjing 210033, China]
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;R446.62
DOI:
10.3969/j.issn.1671-7414.2024.01.023
文献标志码:
A
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-timePCR,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 ~ 250 000 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。
Abstract:
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 ~ 250 000 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.

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备注/Memo

备注/Memo:
基金项目:深圳光明区软科学研究项目(项目批号2021R01097)。
作者简介:夏成静(1976–),男,硕士学位,主管技师,临床检验诊断学,临检专业,E-mail:cjxia1976@163.com。
更新日期/Last Update: 2024-01-15