摘要
为建立一种快速、简便的猪轮状病毒检测方法,采用柠檬酸三钠还原法制备胶体金颗粒,标记纯化的抗猪轮状病毒VP6蛋白单克隆抗体,在硝酸纤维素膜上喷加纯化的抗猪轮状病毒VP6蛋白多克隆抗体和羊抗鼠IgG,制成检测猪轮状病毒抗原的胶体金免疫层析试纸条。结果表明,所制备的试纸条用于检测猪轮状病毒感染的MA104细胞培养物,在检测线处出现红色反应带,未感染病毒的细胞培养物在检测线处未出现反应条带,上述培养物在质控线处均出现红色反应条带;进一步试验表明,试纸条检出病毒培养液(TCID50是10-6.25/0.1 mL)的最低限为1∶32稀释;用不同批次的试纸条重复检测,结果无差异;该试纸条不与相关的腹泻病毒猪传染性胃肠炎病毒、流行性腹泻病毒发生反应,所制备的试纸条具有一定的敏感性和特异性,重复性良好。
To develop a rapid and simple assay for detection of porcine rotavirus （PRV） ,colloidal gold particles labeled with purified monoclonal antibody against PRV VP6 protein were prepared by reduction of sodium citrate. The purified polyclonal antibodies against PRV VP6 protein and goat anti mouse IgG were solidified to nitrocellulose filter to prepare the colloidal gold immunochromatographic strip. MA104 cell cultures infected with PRV and non-infected were detected by the strips, respectively. The results showed that there was a red reaction strip at the test line only when detecting the infected cells, and the red reaction strips all appeared at the control line whatever infected. The test strips detecting virus culture （TCID50：10^-6.25/0.1 mL） had a dilution 1 ： 32,which did not react with transmissible gastroenteritis virus of swine（TGEV） and porcine epidemic diarrhea virus（PEDV）,and the results of different batches of test strips were the same. These results revealed that the test strip has good sensitivity, reproducibility, and specificity.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2008年第11期952-956,共5页
Chinese Veterinary Science
基金
黑龙江省“十一五”科技计划项目(GA06B202-4)
东北农业大学科技创新项目(CX2008)
关键词
猪轮状病毒
胶体金
免疫层析
试纸条
porcine rotavirus
colloidal gold
immunochromatography
test strip
