[1]刁玉梅,宫鹏涛,李巍,等.微小隐孢子虫病毒衣壳蛋白抗体检测ELISA方法的建立[J].中国预防兽医学报,2011,(07):537-540.
 DIAO Yu-mei,GONG Peng-tao,LI Wei,et al.Development of indirect ELISA for detection of the antibody against Cryptosporidium parvum virus[J].Chinese journal of preventive veterinary medicine,2011,(07):537-540.
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微小隐孢子虫病毒衣壳蛋白抗体检测ELISA方法的建立
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2011年07期
页码:
537-540
栏目:
诊断和检测技术
出版日期:
2011-07-15

文章信息/Info

Title:
Development of indirect ELISA for detection of the antibody against Cryptosporidium parvum virus
作者:
刁玉梅;宫鹏涛;李巍;苏利波;黄祥盛;高华义;李赫;胡进平;李建华;张西臣;
吉林大学畜牧兽医学院;
Author(s):
DIAO Yu-mei GONG Peng-tao LI Wei SU Li-bo HUANG Xiang-sheng GAO Hua-yi LI He HU Jin-ping LI Jian-hua ZHANG Xi-chen
College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China
关键词:
微小隐孢子虫dsRNA病毒衣壳蛋白间接ELISA
Keywords:
Cryptosporidium parvum dsRNA virus caspid protein indirect ELISA assay
摘要:
为建立快捷的含病毒隐孢子虫的检测方法,本研究利用已构建的微小隐孢子虫病毒衣壳蛋白重组表达质粒表达并纯化重组蛋白,建立以该重组衣壳蛋白为包被抗原检测抗体的间接ELISA检测方法。经优化后间接ELISA的最佳条件为:抗原包被浓度为0.25μg/孔,被检血清稀释度为1∶200,酶标二抗稀释度为1∶2000,封闭液为含5%脱脂奶粉的PBST溶液。该方法检测安氏隐孢子虫阳性血清,柔嫩艾美尔球虫阳性血清,蓝氏贾第虫阳性血清,弓形虫阳性血清与微小隐孢子虫阳性结果均为阴性。该方法灵敏度为1∶1600,特异性较强且具可重复性,可用于含病毒隐孢子虫的抗体检测及流行病学调查。
Abstract:
To develop an indirect ELISA for detection of the antibody against Cryptosporidium parvum virus(CSpV), recombinant protein of CSpV capsid protein was purified as coating antigen. The optimal antigen concentration for coating was 0.25 μg each well, and the optimal dilutions of the sera of C. parvum and enzyme-labeled the second antibody were 1∶200 and 1∶2 000 respectively, blocking agent was the 5% skim milk PBST by chessboard titration. The developed indirect ELISA had no cross reaction with positive mouse serum against C. parvum, C. andersoni, E. tenella and T. gondii. It showed high sensitivity of 1∶1 600. The indirect ELISA for detection of the antibody against C. parvum virus might be used for the diagnosis and epidemiological investigation of the virus infected Cryptosporidium

参考文献/References:

[1] Hayes M B,et al. Large community outbreak of cryptosporidiosis due to contamination of a filtered public water supply. The New England Journal of Medicine. 1989
[2] MacKenzie WR,Hoxie NJ,Proctor ME,et al. A massive outbreak in Milwaukee of Cryptosporidium infection transmitted through the public water supply. . 1994
[3] D’ Antonio RG,Winn RE,Taylor JP. A waterborne outbreak ofcryptosporidiosis in normal host. Annals of Internal Medicine. 1985
[4] Nikolai V K,Keith M W,Michael V N,et al. Virus-like double stranded RNAs in the parasitic protozoan Cryptosporidium parvum[J]. Molecular Microbiology. 1997
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[6] Kniel K E,Higgins J A,Trout J M,et al.. Characterization and potential use of a Cryptosporidium parvum virus(CPV)antigen for detecting C.parvum oocysts[J].. Journal of Microbiological Methods. 2004
[7] 朱惠丽,张龙现,宁长申,党海亮,李家诚. 人兽共患隐孢子虫种类及基因型[J]. 寄生虫与医学昆虫学报. 2007年01期
[8] 蒋正军,马世东,蔡丽娟,单文鲁,王树双,尹燕博,龚振华,李葳,宣华. 非洲猪瘟间接ELISA诊断试剂盒的研究[J]. 中国预防兽医学报. 2000年S1期

备注/Memo

备注/Memo:
基金:国家科技支撑项目(2007BAD40B05);;吉林省科技发展计划项目(20100160)
更新日期/Last Update: 2011-09-29