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当前位置: 首页 > 文章 > H5亚型禽流感病毒荧光检测试纸条的研制 中国兽医杂志 2019 (6) 32-35,132
Position: Home > Articles > H5亚型禽流感病毒荧光检测试纸条的研制 Chinese Journal of Veterinary Medicine 2019 (6) 32-35,132

H5亚型禽流感病毒荧光检测试纸条的研制

作  者：
孙洁;陈雨;吴峰;王津津;陈兵;郑晓聪;陶虹;刘建利;陈书琨;林庆燕;秦智锋
单  位：
清华大学深圳研究生院;深圳市检验检疫科学研究院;大连海洋大学水产与生命学院;深圳海关动植物检验检疫技术中心
关键词：
H5亚型禽流感病毒;荧光量子点;检测试纸条;快速检测
摘  要：
为了对引起家禽重大经济损失和具有公共卫生意义的H5亚型禽流感病毒(AIV)进行快速超敏检测,本研究研制了针对HA蛋白的4株单克隆抗体(MAb)杂交瘤细胞株,通过MAb的两两配对试验,以荧光量子点作为抗体标记生物材料,基于夹心法原理建立了H5亚型AIV荧光免疫层析检测试纸条.结果显示,该方法能够特异性检测出H5N1、H5N2等H5亚型AIV,而与H1N1、H3N2、H7N9、H9N2、NDV等其他病原无交叉反应;对标准检测抗原H5N1(Re-6)的最低检测量为1 ng/mL,比市售H5亚型AIV胶体金检测试纸条灵敏度高100倍;该方法的变异系数(CV)值在10%以内,与商品化的试剂盒检测结果符合率达99.8%.本文建立的H5亚型AIV荧光检测试纸条快速、敏感,检测结果准确可靠,为H5亚型禽流感的流行病学调查和快速诊断提供了基础.
作  者：
SUN Jie;CHEN Yu;WU Feng;WANG Jin-jin;CHEN Bing;ZHENG Xiao-cong;TAO Hong;LIU Jian-li;CHEN Shu-kun;LIN Qing-yan;QIN Zhi-feng;Animal & Plant Inspection and Quarantine Technology Center of Shenzhen Customs;Shenzhen Academy of Inspection and Quarantine;Dalian Ocean University College of Fisheries and Life Science;Graduate school at Shenzhen,Tsinghua University;
关键词：
H5 Avian influenza virus;;quantum dots;;lateral-flow immunoassay;;rapid detection
摘  要：
In order to detect H5 subtype of Avian influenza virus(AIV) sensitively and specifically on site where H5 AIV causes large loss on avian business and public health, four hybridoma cell strains secreting monoclonal antibodies(MAb) against HA protein were explored, and a fast and ultrasensitive lateral-flow immunoassay strip with quantum dots(QDs) as an novel detection report probe(H5-AIV-QD-LFIA) was established against H5 subtype AIV based on double antibodies sandwich principle. The H5-AIV-QD-LFIA was specific for testing H5 N1、H5 N2,etc, but had no false positive results with H1 N1,H3 N2,H7 N9,H9 N2 and NDV ect. The sensitivity of the H5-AIV-QD-LFIA is 1 ng/mL for standard H5 N1(Re-6)strain and was 100 time higher than colloidal gold strip for H5 virus on sale. The repeatability of the assay was acceptable with CV≤10% and the consistency was 98% with commercial kits. The developed H5-AIV-QD-LFIA was a promising method for monitor and surveillance of H5 subtype virus.

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