Development of a blocking ELISA for detection of serum neutralizing antibodies against porcine circovirus type 2
Development of a blocking ELISA for detection of serum neutralizing antibodies against porcine circovirus type 2
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开发用于检测猪圆环病毒 2 型血清中和抗体的封闭 ELISA
DOI：
10.1016/j.jviromet.2010.09.023
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发表时间：
2011-01-01
影响因子：
3.1
通讯作者：
Liu, Changming
中科院分区：
文献类型：
--
作者：
Huang, Liping;Lu, Yuehua;Liu, Changming
建立了检测猪圆环病毒2型（PCV2）血清中和抗体的单克隆抗体阻断ELISA方法。用PCV2重组衣壳蛋白免疫昆虫细胞制备的具有中和活性的单克隆抗体作为检测抗体。与血清中和试验进行比较，其灵敏度和特异性分别为98.8%和88.5%。阻断ELISA与血清中和试验结果呈显著正相关（r = 0.9381）。通过检测实验和商业猪血清验证了该测定法。纵向抗体谱显示，接种后2周检测到血清中和抗体，4周时检出率达到100%。血清中和抗体谱显示从4周龄到13周龄的降低，并且在来自商业猪场的猪中在13周龄后血清转化。对9个商品猪场的703份血清进行检测，阳性检出率为73%。该方法简便、特异、灵敏、简便，可用于PCV2血清中和抗体的流行病学调查和评价。(C)2010 Elsevier B.V.保留所有权利。
A monoclonal antibody (Mab)-based blocking ELISA was developed for the detection of serum neutralizing antibodies to porcine circovirus type 2 (PCV2). The Mab with neutralizing activity, which was produced by immunizing a recombinant capsid protein of PCV2 expressed in insect cells, was used as the detector antibody. The assay was evaluated in comparison with a serum neutralization assay, and its sensitivity and specificity were determined to be 98.8% and 88.5%, respectively. A significant positive correlation was found between results of the blocking ELISA and the serum neutralization assay (r = 0.9381). The assay was verified by testing experimental and commercial pig sera. A longitudinal antibody profile showed that serum neutralizing antibodies were detected 2 weeks after vaccination and that the detection rate reached 100% at 4 weeks. The serum neutralizing antibody profile showed a decrease from the age of 4 to 13 weeks, and seroconversion after 13 weeks in pigs from a commercial pig farm. Additionally, the positive detection rate in 703 sera collected from nine commercial pig farms was 73%. This report demonstrates that the assay is a simple, specific, sensitive and convenient method for epidemiological surveys and evaluations of serum neutralizing antibodies against PCV2. (C) 2010 Elsevier B.V. All rights reserved.