IFA和ELISA检测J亚群禽白血病病毒的比较研究

Comparison of IFA and ELISA for Detection of Avain Leukosis Virus Subgroup J in DF1 Cell Cultures

为比较3种禽白血病病毒（ALV）抗原检测ELISA试剂盒的特异性和灵敏度,将J亚群禽白血病病毒（ALV-J）传染性克隆rNX0101株以病毒原液（9×103 TCID50）、10倍病毒稀释液（9×102 TCID50）、100倍病毒稀释液（90 TCID50）接种培养于6孔板的DF1细胞,每孔均提前放入灭菌的细胞爬片,于接种后第1～6天每天取上清用3种ELISA试剂盒（A、B、C）检测ALV p27抗原,同时在接种后的第3天和第6天取出细胞爬片进行间接免疫荧光法（IFA）检测.结果表明,IFA方法对高、中、低剂量接种3d后均可检出ALV-J的感染,而ELISA试剂盒中只有A试剂盒在高剂量接种时才能检出,试剂盒B和试剂盒C最早要在第4天才可捡出;中、低剂量接种,3种试剂盒在第4～5天才能检出;第6天时,由于病毒的明显复制,无论IFA方法还是3种ELISA试剂盒都可检出ALV-J的感染.本研究结果为正确选择商品化禽白血病抗原检测试剂盒提供了借鉴.

To evaluate three ELISA kits for avain leukosis virus （ALV）,an infectious clone rNX0101 of ALV subgroup J （ALV-J）was inoculated into DF1 ceils with three different doses （9×103,9×102,90 TCIDs0 respectively）and the supernatants of DF1 cell cultures were obtained at 1 to 6 days post inoculation and tested with A,B,C kits. The infected cells were detected by indirect immunofluorescence assay （IFA）at 3 and 6 days post inoculation respectively.The results showed that infected cells at 3 days and 6 days post inoculation all could be identified by IFA after inoculated with different dose,while only ELISA kit A could detect p27 antigen at 3 days post inoculation with the high dose and none of inoculated ceils was detected positively with B and C ELISA kits at the same time. All samples infected by different doses showed positive to ALV-J with both ELISA kits and IFA 6 days post inoculation. The results indicated that sensitivity among A, B and C ELISA kits was different in the detection of ALV-J with three doses of inoculation and IFA was more sensitive than commercial ELISA kits.