3种ELISA试剂盒检测不同亚型外源性鸡白血病病毒的比较

Comparision of Dynamic Detection with Three ELISA Kits to Different Subgroups of Exosomic ALVs in DF1 Cells

为建立一种稳定、快速从感染鸡体内检测或分离外源性鸡白血病病毒(ALV)的简易方法,作者使用A亚型(ALV-A)、C亚型(ALV-C)以及2株J亚型(ALV-J-PY和ALV-J-WS)鸡白血病病毒(ALV)按高、中、低(即100、10、1μL)3种接种量人工接种DF1细胞,在接种后不同时间用A、B、C 3种ELISA试剂盒检测ALV抗原(P27)。结果表明,对ALV-A和ALV-J-PY,高剂量接种时,用A试剂盒在接种后第3天即可检测出;低剂量接种时,第7天可检出。使用B试剂盒,2株病毒的检出时间延长(高剂量组分别为第7天和第5天;低剂量组分别为第15天和第13天);使用C试剂盒,所需检出时间最长(接种后第13天)。对ALV-C和ALV-J-WS毒株,A试剂盒对高剂量组,分别能够在接种后第5天和第9天检出,其它中、低接种量组15 d内均没有检出;而B、C2个试剂盒对3个接种剂量组均没有检测出。从以上结果看出,3种ELISA试剂盒对3种亚型ALV毒株的检出时间存在明显不同,A试剂盒灵敏度最高,能最早作出检测;B试剂盒灵敏度次之。同时,无论使用哪种试剂盒ALVs的检出时间与病毒接种量间存在很大的相关性。本研究结果为外源性ALV的检测及病毒分离研究提供一定的科学依据。

DF1 cells were inoculated with three different doses （100, 10, 1 μL） of ALV-A, ALV- C, ALV-J-PY and ALV-J-WS separated from different chickens to evaluate three kinds of A, B, C ELISA kits. The samples of DF1 cells supernatant were obtained at different days post inocula- tion （dpi） and detected with three kinds of kits after freezing and thawing once. The results indicated that the samples inoculated with ALV-A or ALV-J-PY were both detected positively firstly at the 3rd dpi with A ELISA kit for 100μL inoculation doses and at the 7th dpi for 1μL dose; If with B ELISA kit, two ALVs of subgroup were detected positively within some days post inoculation, but the detectable time were longer than that of A ELISA kit; If with C ELISA kit, the detectable time of viruses were the longest. ALV-C and ALV-J-WS were detected positively with A ELISA kit at the 5th and 9th dpi for 100 μL doses, respectively; but for the other doses, no positive samples were detected at whole observed days. If with B and C ELISA kits, none of inoculated cells with three doses of inoculation was detected positively. It can be concluded that it is dramatically different between A, B and C ELISA kits in the detection of three exosomic sub groups of ALVs The A ELISA kit is more sensitive than other two ELISA kits （B and C）. Simultaneously, the positively detected days to ALVs are related to the doses of inoculation in DF1 cells. The results can be helpful for the application of three ELISA kits to the detection or isolation of exosomic ALVS.