鸡新城疫免疫胶体金诊断技术的研究

The Research of Gold-Immunochromatography for Diagnosis of Chicken Newcastle Disease

为了建立一种快速、简便的检测鸡新城疫病毒(NDV)的胶体金免疫层析法(GICA),采用柠檬酸钠还原法制备胶体金颗粒,标记纯化的ND抗体,并包被在玻璃纤维素膜上,另外将纯化的ND抗体和纯化的兔抗鸡抗体包被在硝酸纤维素膜上,组装成ND快速检测试纸条.用本试验研制的ND快速检测试纸条检测收集的24份样品,其结果与HA结果一致,符合率100%;当病毒HA效价在1∶40以上时,试纸条均能检测为阳性;ND快速检测试纸条与已知的马立克病(MD)病料和鸡传染性法氏囊病(IBD)病料的上清液无交叉反应出现,且保存6个月后重复性100%.试验证明,建立的GICA是一种快速、灵敏、特异的抗原检测方法,可用于鸡新城疫诊断,具有广泛应用价值.

The objective was to develop a rapid and simple colloidal gold immunochromatographic assay （GICA） for the diagnostic of chicken NDV. Tri-sodium citrate with aqueous gold chloride were warmed up and mixed to make gold sol. Colloidal golds were coupled with the purified anti-ND antibody. The gold-anti-ND were wrapped onto glass fibre membrane, and the purified anti-ND and the purified rabbit-anti-chicken antibody were wrapped onto nitrocellulose membrane to prepare a kit. 24 sample of dejection were detected for ND-Ag by GICA and HA. The results were correspondent and the corresponding ratio was 100%. When the HA titer was above 1 ： 40, the GICA result was positive. Detecting the sample of MD and IBD, the GICA result was negative. The repetition ratio was 100% after the kit had been stored for 6 months. It was confirmed that the method was sensitive, specific, rapid and useful for the detection of ND-Ag.