[1]江地科,尹清清,项明源,等.检测猪瘟病毒胶体金和量子点试纸条的初步研制[J].江苏农业学报,2020,(01):116-121.[doi:doi:10.3969/j.issn.1000-4440.2020.01.016]
 JIANG Di-ke,YIN Qing-qing,XIANG Ming-yuan,et al.Development of colloidal gold and quantum dot test strip for detection of classical swine fever virus[J].,2020,(01):116-121.[doi:doi:10.3969/j.issn.1000-4440.2020.01.016]
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检测猪瘟病毒胶体金和量子点试纸条的初步研制()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2020年01期
页码:
116-121
栏目:
畜牧兽医·水产养殖
出版日期:
2020-02-29

文章信息/Info

Title:
Development of colloidal gold and quantum dot test strip for detection of classical swine fever virus
作者:
江地科12尹清清12项明源12王印12姚学萍1罗燕1杨泽晓1张鹏飞12张保海12周丽军12
(1.四川农业大学动物医学院,四川成都611130;2.动物疫病与人类健康四川省重点实验室,四川成都611130)
Author(s):
JIANG Di-ke12YIN Qing-qing12XIANG Ming-yuan12WANG-Yin12YAO Xue-ping1LUO-Yan1YANG Ze-xiao1ZHANG Peng-fei12ZHANG Bao-hai12ZHOU Li-jun12</
(1.College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China;2.Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu 611130, China)
关键词:
猪瘟病毒E2蛋白胶体金量子点
Keywords:
classical swine fever virusE2 proteincolloidal goldquantum dots
分类号:
S852.65+1
DOI:
doi:10.3969/j.issn.1000-4440.2020.01.016
文献标志码:
A
摘要:
为建立快速、简便检测猪瘟病毒(CSFV)抗体的方法,本研究采用胶体金和量子点免疫层析技术,将纯化的重组CSFV E2蛋白作为捕捉抗原,以纯化的抗CSFV E2蛋白多克隆抗体和HRP兔抗猪IgG分别包被于硝酸纤维素膜上,作为质控线(C线)和检测线(T线),优化反应条件,制备免疫层析试纸条,用于临床检测。结果表明,制备的2种试纸条都可用于检测CSFV标准阳性血清,使用不同批次试纸条进行检测,结果无差异。2种试纸条与CSFV阳性血清反应呈阳性,与伪狂犬病毒(PRV)、猪乙型脑炎病毒(JEV)、猪繁殖与呼吸综合征病毒(PRRSV)和圆环病毒2型(PCV2)反应呈阴性;胶体金试纸条在临床样品中阳性检出率为84.38%(81/96),与CSFV抗体检测ELISA试剂盒检测阳性符合率为92.05%(81/88);量子点试纸条阳性检出率为87.50%(84/96),与ELISA试剂盒检测阳性符合率为95.45%(84/88)。本研究制备的2种试纸条都具有较好的特异性、敏感性、重复性和稳定性。
Abstract:
Two test strips for rapid detection of the classical swine fever virus(CSFV) antibody were developed based on colloidal gold and quantum dots immunochromatographic assay. The recombinant CSFV E2 protein was incubated on test strip as capture antigen, and the rabbit anti-pig IgG was immobilized on the test line as detection antibody. In addition, anti-CSFV E2 protein polyclonal antibody was blotted on the control line of the nitrocellulose membrane. The results showed that the two test strips could be used to detect the CSFV positive serum. The test strips did not capture unspecific virus proteins which derived from pseudorabies virus(PRV), Japanese encephalitis virus(JEV), porcine reproductive and respiratory syndrome virus(PRRSV) and porcine circovirus type 2(PCV2). In the detection of clinical samples, the positive detection rate of colloidal gold immunochromatographic strip was 84.38% (81/96), and the coincidence rate was 92.05%(81/88) compared with CSFV antibody IDEXX kit (ELISA). The positive detection rate of quantum dot immunochromatographic strip was 87.50%(84/96), and the coincidence rate was 95.45%(84/88)compared with CSFV antibody IDEXX kit (ELISA). The two test strips prepared in this study have good specificity, sensitivity, repeatability and stability.

参考文献/References:

[1]YUAN J, ZHU M, DENG S, et al. Classical swine fever virus induces pyroptosis in the peripheral lymphoid organs of infected pigs[J]. Virus Research, 2018,250(5):37-42.
[2]陈姝承,孙慧敏,李素,等.针对猪瘟病毒E2蛋白的嵌合猪源化单克隆抗体的表达及抗病毒活性鉴定[J].生物工程学报,2017,33(8):1235-1243.
[3]XU X G, TONG D W, LIU H W. Baculovirus surface display of E0E1E2 envelope glycoprotein of CSFV and immunogenicity of the displayed proteins in mouse model[J]. Journal of Northwest A & F University,2009,37(12):73-79.
[4]GONG Y, TROWBRIDGE R, MACNAUGHTON T B, et al. Characterization of RNA synthesis during a one-step growth curve and of the replication mechanism of bovine viral diarrhoea virus[J]. Journal of General Virology, 1996, 77(11):2729-2736.
[5]孙雪霏. 猪弓形虫抗体间接ELISA方法的建立及胶体金试纸条的制备[D].长春:吉林农业大学, 2016.
[6]傅秋玲,程龙飞,万春和,等.半番鸭胚中鹅细小病毒的分离鉴定及其基因组分子特征分析[J].畜牧兽医学报, 2017, 48(11):2148-2156.
[7]KOBAYASHI H, HAMA Y, KOYAMA Y, et al. Simultaneous multicolor imaging of five different lymphatic basins using quantum dots[J]. Nano Letters, 2007, 7(6):1711-1716.
[8]QIU W, XU H, TAKALKAR S, et al. Carbon nanotube-based lateral flow biosensor for sensitive and rapid detection of DNA sequence[J].Biosensors and Bioelectronics,2015, 64(2):367-372.
[9]LIU C Y, JIA Q J, YANG C H, et al. Lateral flow immunochromatographic assay for sensitive pesticide detection by using Fe3O4 nanoparticle aggregates as color reagents[J]. Analytical Chemistry,2011,83(17):6778-6784.
[10]王向鹏,孙元,杨增岐,等.检测猪瘟病毒野毒株胶体金免疫层析方法的建立[J].中国预防兽医学报,2010,32(6):441-445.
[11]魏后军,范志宇,王芳,等.检测兔出血症病毒抗体胶体金试纸条的研制及初步应用[J].畜牧兽医学报,2018,49(3):614-619.
[12]TARRADAS J, EUGENIA D L T M, ROSELL R, et al. The impact of CSFV on the immune response to control infection[J].Virus Research,2014,185(6):82-91.
[13]韩焘,张宁宁,齐鲁,等.猪瘟退出强制免疫后的若干问题[J].中国动物检疫, 2017(5):60-63.
[14]袁莉,杨顺利,尚佑军,等.猪瘟病毒结构蛋白抗体间接ELISA检测方法的建立及评价[J].西北农业学报,2018,27(9):43-49.
[15]蔺辉星,周红,童泽鑫,等.猪瘟病毒血清抗体间接ELISA检测方法的建立[J]. 中国兽医科学,2017(1):9-15.
[16]张杰,胡军,刘杨,等.肺炎支原体快速检测方法的建立及应用评价[J].国际检验医学杂志,2016,37(17):2379-2381.
[17]樊淑华,王永立.胶体金免疫层析技术应用研究进展[J].动物医学进展,2014,35(10): 99-103.
[18]胡纪文,张大准,熊建辉,等.肺炎支原体抗原量子点免疫层析法的建立及初步应用[J].国际检验医学杂志,2018,39(2):220-223.
[19]尹梅.猪瘟病毒抗体胶体金免疫层析快速检测试纸条的研制及初步应用[D].南京:南京农业大学,2013.

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备注/Memo

备注/Memo:
收稿日期:2019-05-28基金项目:国家农业产业技术体系四川兽药创新团队专项(CARS-SVDIP)作者简介:江地科(1993-),男,四川乐山人,硕士研究生,主要从事动物传染病病原微生物研究。(E-mail) 1192828928@qq.com。尹清清为共同第一作者。通讯作者:王印,( E-mail)yaanwangyin@tom.com
更新日期/Last Update: 2020-03-13