不同检测技术在K亚群禽白血病病毒检测中的应用

Application of Different Detection Techniques in Detecting Subgroup K Avian Leukemia Virus

禽白血病(avian leukosis,AL)以垂直传播为主,被列为我国种禽场必须要实现净化的主要禽病之一,通过1日龄雏鸡胎粪检测淘汰等程序是实施AL净化的主要步骤之一。K亚群禽白血病病毒(ALV-K)是近年来在黄羽肉鸡群中流行的主要亚群。为比较不同检测方法对ALV-K的灵敏度差异,将ALV-K经卵黄囊接种感染SPF鸡胚后构建垂直感染模型,对1日龄雏鸡采集胎粪和抗凝血,接种DF-1细胞后进行病毒分离,维持7 d收取细胞上清。收集胎粪以及胎粪和抗凝血的病毒分离细胞上清等样品构建样品盘,分别以磁微粒化学发光检测试剂盒、3家公司ELISA试剂盒、胶体金检测试纸条等进行检测。结果显示,确定样品盘利用不同检测方法的阳性检出率均为100%,各检测技术检出结果一致;但不同方法的灵敏度有差异,其中磁微粒化学发光技术灵敏度优势明显,可更好地判定ALV垂直传播状态。本研究提出了一种可评估不同检测技术灵敏度的良好模型,为进一步理解不同检测技术用于AL净化的价值提供了参考。

Avian leukemia(AL),mainly with vertical transmission,is listed as one of the major poultry diseases that must be purified in breeding poultry farms in China by key steps including elimination through detection of faeces of 1-day-old chicks.Subgroup K avian leukemia virus(ALV-K)is one of the major subgroups prevalent in yellow-feathered broilers in recent years.In order to compare the sensitivity difference of different detection methods for ALV-K,a vertical infection model was constructed through inoculating ALV-K into yolk sac of SPF chicken embryos,samples of faeces and anticoagulant blood were collected from 1-day-old chicks,and inoculated into DF1 cells for viral isolation,after 7 days,cell supernatant was collected.Samples of faeces as well as the cell supernatant after virus isolation of faeces and anticoagulant blood were collected to construct a sample disk,which were detected by magnetic particle chemiluminescence,ELISA kits made by three manufacturers and colloidal gold test strips,respectively.he results showed that the positive detection rates of different methods were all 100%,indicating consistency,but different sensitivity,magnetic particle chemiluminescence was obviously dominant,and could be used to better determine the vertical transmission status of ALV.In conclusion,a good model was provided to evaluate the sensitivity of different detection techniques,providing a reference for further research on the value of different detection techniques for AL purification.