【作者】 戴荣四；

【导师】 刘毅；

【作者基本信息】 湖南农业大学 ， 基础兽医学， 2004， 硕士

【摘要】 鸡新城疫是鸡的一种高度感染和致死性疾病，是由一种侵害呼吸道、胃肠道以及中枢神经系统的新城疫病毒(NDV)所引起的。病毒存在于病鸡的血液、脑、内脏、粪便和口鼻中，国内外对本病的控制仍没有完全解决，鸡新城疫对养鸡业有着巨大的威胁。所以有必要建立一种快速、简便、准确的胶体金免疫层析法(GICA)用于鸡新城疫的诊断。胶体会技术最早是Faulk等于1971年报道的应用于电镜检查技术的一种示踪技术。与传统方法比较，由于胶体金具有肉眼可观察的红色，所以以这种技术为基础开发的检测试剂盒有快捷、准确、高特异性、高敏感性的特点，灵敏度可达0.5～1ng/mL，适用于各级医院、卫生所、家庭个人诊断、保健、体检。目前以此方法为基础发展起来的检测试剂盒涉及范围相当广泛，包括HCG等激素、肿瘤标志物、心血管标志物、传染病的抗原与抗体的检测等。采用柠檬酸钠还原法制备胶体金颗粒，标记纯化的鸡新城疫抗体，并包被在玻璃纤维素膜上，另外将纯化的鸡新城疫抗体和纯化的兔抗鸡抗体包被在硝酸纤维素膜上，组装成鸡新城疫快速诊断试纸条。当检样中含有鸡新城疫病毒抗原时，试纸条上出现两条红线，为阳性；当检样中不含有鸡新城疫病毒抗原时，试纸条出现一条红线，为阴性。用本试验研制的鸡新城疫快速诊断试纸条检测收集的20份样品，其结果与HA结果相对应，符合率100％。当病毒HA效价在1：32以上时，试纸条均能检测为阳性。鸡新城疫快速诊断试纸条与已知的禽流感病料和鸡传染性支气管炎病料的上清夜无交叉反应出现。进行热稳定性试验稳定性好。试验证明，本研究建立的胶体金免疫层析法(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，也可用于鸡新城疫的流行病学调查，具有推广价值。更多还原

【Abstract】 Developing a rapid kit for the diagnostic of Chicken Newcastle disease Chicken Newcastle Disease is a highly infectious and deadly disease, it is caused by NDV (Newcastle disease virus) that violate the respiratory tract the digestive tract and the centrencephalic system. The viruses are in blood, brain, internal organs excrement mouth and nose, there is no complete solution for this disease at home and abroad, so it is a big menace for chicken cultivating industry. So it is necessary to establish a rapid, simple,self-performing GICA(gold immunochromatographic assay) for the diagnosis of Chicken Newcastle Disease. Colloidal gold technique firstly was a kind of the tracking technique , which was reported to apply to electron microscopy(IEM) detecting technique by persons like Faulk in 1971. By comparison with the traditional technique, since the colloidal gold embodies the property of red to be seen by nake eyes, so the detective test box developed basically on this technique has rapid, sensitive, special, sensitive features, colloidal golds were coupled with Chicken Newcastle Disease to prepare an GICA apparatus, marking the purified antibody of Chicken Newcastle Disease, wrapping them onto nitrocellose strips, and then wrapping themand the purified rabbit antibody for chicken onto nitrocellose, assembled a rapid kit for the diagnostic of Chicken Newcastle Disease. If the sample contains virus antigen of Chicken Newcastle Disease and the kit appears two red lines, it is positive; if the sample contains no virus antigen of Chicken Newcastle Disease and the kit appears one red line, it is negative. When using the rapid diagnosis of kits for Chicken Newcastle Disease, which are developed in this experiment, to detect the accumulated 20 samples, their results correspond the result of HA, and the corresponding rate is 100%, When virus HA titer is above 1:32, the kits are all positive. The rapid diagnosis of kit for Chicken Newcastle Disease appears no intercross reaction with the clear liquid in Avian Ingluenza(AI) and Chicken Infectious Bronchitis(CIB). It was confirmed that the GICA(gold immunochromatographic assay) established in this experiment is a kind of rapid, senstive, and special antigen test method, it can be used to test Chicken Newcastle Disease, and to investigate epidemiology of Chicken Newcastle Disease, and has the value to be generalized.更多还原