猪瘟病毒抗体纳米荧光检测试纸条方法建立及性能评价

doi:10.13560/j.cnki.biotech.bull.1985.2019-0986

生物技术通报 ›› 2020, Vol. 36 ›› Issue (5): 74-79.doi: 10.13560/j.cnki.biotech.bull.1985.2019-0986

• 生物分析方法与标准物质专题（专题主编：李亮副研究员） • 上一篇下一篇

猪瘟病毒抗体纳米荧光检测试纸条方法建立及性能评价

吴昊星, 刘百红, 刘雪微, 周景云, 马永缨, 杨欣艳, 李宝春, 陈西钊

北京世纪元亨动物防疫技术有限公司,北京 100089

收稿日期:2019-10-17 出版日期:2020-05-26 发布日期:2020-06-03
作者简介:吴昊星,男,研究方向：动物疫病检测;E-mail：wuhaoxing@anheal.com
基金资助:
国家重点研发计划基金项目（2018YFD500801）

Establishment and Performance Evaluation of Nano-fluorescence Strip for Detecting CSFV Antibody

WU Hao-xing, LIU Bai-hong, LIU Xue-wei, ZHOU Jing-yun, MA Yong-ying, YANG Xin-yan, LI Bao-chun, CHEN Xi-zhao

Beijing Anheal Laboratories Co. Ltd,Beijing 100089

Received:2019-10-17 Published:2020-05-26 Online:2020-06-03

摘要/Abstract

摘要： 基于镧系元素Eu微球标记技术建立了一种猪瘟病毒抗体检测的免疫层析方法。通过对反应体系中包被浓度、复溶浓度以及反应时间等因素进行优化,确定最适的反应条件,建立检测方法,然后通过从敏感性、特异性、重复性、临床评价等方面对其进行性能评价。对反应体系进行优化,最终确定包被浓度为0.1 mg/mL,复溶浓度为6倍稀释,检测时间为15 min。通过对试纸条的性能评价可以得出,猪瘟病毒抗体荧光检测试纸条的敏感性为猪瘟阳性血清国家参考品倍比稀释至1∶128倍仍可以检测到;对常见的猪繁殖与呼吸综合征、猪I型疱疹病毒、猪口蹄疫、猪圆环病毒2型、猪流行性腹泻、牛病毒性腹泻病毒、羊边界病毒等抗体阳性血清无交叉反应;批内和批内变异系数均小于10%;经临床评价,与商品化的猪瘟病毒抗体ELISA检测试剂盒相比阳性符合率为90%,阴性符合率为100%,总符合率为97.7%;与荧光抗体中和试验（FVNT）的阴性符合率为100%,阳性符合率为93%,总符合率为98.5%。综合评定认为本研究建立的猪瘟病毒抗体纳米荧光检测方法,符合各项性能参数,可以快速、经济、方便地对猪个体及群体进行猪瘟病毒抗体检测评估,可广泛应用于猪场管理中。

关键词: 纳米荧光, 猪瘟病毒, 荧光免疫层析

Abstract: An immunochromatography method for the detection of the antibodies against classical swine fever virus（CSFV）was established based on lanthanide europium microsphere labeling technology. By optimizing the coating concentration,resolving concentration and reaction time in the reaction system,the optimal reaction conditions were determined and the detection method was established. Then the performance of the system was evaluated from sensitivity,specificity,precision and clinical evaluation. The optimized reaction system was as that the concentration of coating was at 0.1 mg/mL,the dilution parameter was 6 times,and the detection time was 15 min. From the performance evaluation of the strips,the sensitivity of the strip for the detection of CSFV antibody was that the diluted 128 times of national reference samples of swine fever positive serum were still detected,and no cross-reactions were observed with the positive sera against common CSFV,for example PPRSV,PRV,FMDV,PCV,PEDV,BVDV,and BDV. The coefficient of variation was < 10%. Compared with the commercial ELISA kit of CSFV antibody,the coincidence rate for positive was 90% and that for negative was 100%,and the total coincidence rate was 97.7%. Compared with FVNT,the coincidence rate for positive were 93% and that for negative was 100%,and the total coincidence rate was 98.5%. According to the above comprehensive evaluation,the fluorescence detection method of CSFV antibody established in this study conforms to all the performance parameters. It can be used for rapid,economical and convenient detection of CSFV antibody in pigs,and can be widely applied to pig farm management at the grass-roots units.

Key words: nano-fluorescence, classical swine fever virus, fluorescence immunochromatography

吴昊星, 刘百红, 刘雪微, 周景云, 马永缨, 杨欣艳, 李宝春, 陈西钊. 猪瘟病毒抗体纳米荧光检测试纸条方法建立及性能评价[J]. 生物技术通报, 2020, 36(5): 74-79.

WU Hao-xing, LIU Bai-hong, LIU Xue-wei, ZHOU Jing-yun, MA Yong-ying, YANG Xin-yan, LI Bao-chun, CHEN Xi-zhao. Establishment and Performance Evaluation of Nano-fluorescence Strip for Detecting CSFV Antibody[J]. Biotechnology Bulletin, 2020, 36(5): 74-79.

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猪瘟病毒抗体纳米荧光检测试纸条方法建立及性能评价

Establishment and Performance Evaluation of Nano-fluorescence Strip for Detecting CSFV Antibody

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