用于检测抗猪细小病毒抗体的重组病毒粒子酶联免疫吸附测定法的建立与评估

Development and evaluation of the rVP-ELISA for detection of antibodies against porcine parvovirus.

作者信息

Kong Miaomiao, Peng Yonggang, Cui Yuchao, Chang Tiecheng, Wang Xiaoling, Liu Zhaoxia, Liu Yonggang, Zhu Yu, Luo Yakun, Tang Qinghai, Feng Li, Cui Shangjin

机构信息

State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, 427 Maduan Street, Nangang District, Harbin 150001, Heilongjiang, China; Institute of Animal Sciences (IAS), Chinese Academy of Agricultural Sciences, Beijing 100094, China.

出版信息

J Virol Methods. 2014 Sep;206:115-8. doi: 10.1016/j.jviromet.2014.06.008. Epub 2014 Jun 16.

DOI:10.1016/j.jviromet.2014.06.008

PMID:24945904

Abstract

The gene encoding the VP2 protein of porcine parvovirus (PPV) was expressed in an insect-baculovirus system. The recombinant (r) VP2 was similar antigenically/functionally to the native capsid protein as demonstrated by hemagglutination (HA), Western blotting using PPV positive sera. The purified rVP2 proteins were used as coating antigen to establish a rVP-ELISA method for detection of PPV positive and negative sera from pigs. The optimal operating conditions of the rVP-ELISA were: the concentration of rVP2 proteins coated on the wells was 2 μg/mL; the diluted concentration of serum was 1: 150 and that of the enzyme-labeled antibody was 1: 6000. A total of 596 sera were detected by this assay, and the average positive rate was 87%. Compared with France LSI kit, the result showed that the coincidence rate was 96.7%. In conclusion, the rVP2-ELISA is a sensitive and specific method for detecting antibodies against PPV.

摘要

猪细小病毒（PPV）的VP2蛋白编码基因在昆虫-杆状病毒系统中得以表达。重组（r）VP2在抗原性/功能上与天然衣壳蛋白相似，这通过血凝试验（HA）以及使用PPV阳性血清的蛋白质印迹法得以证实。纯化后的rVP2蛋白被用作包被抗原，以建立一种用于检测猪PPV阳性和阴性血清的rVP - ELISA方法。rVP - ELISA的最佳操作条件为：包被于孔中的rVP2蛋白浓度为2μg/mL；血清稀释浓度为1:150，酶标抗体稀释浓度为1:6000。通过该检测方法共检测了596份血清，平均阳性率为87%。与法国LSI试剂盒相比，结果显示符合率为96.7%。综上所述，rVP2 - ELISA是一种检测PPV抗体的灵敏且特异的方法。