摘要
为比较不同方法在测定J亚群禽白血病病毒(ALV-J)组织细胞半数感染量(TCID50)上的差异,将ALV-J传染性克隆r NX0101株同一病毒保存液以10倍梯度稀释后按照常规方法分别接种已长满DF-1细胞和鸡胚成纤维细胞(CEF)的96孔板,维持7 d后取所有细胞培养上清以ALV-p27抗原检测试剂盒检测p27抗原,细胞固定后以ALV-J特异性单克隆抗体JE9进行间接免疫荧光(IFA)检测,确定病毒感染孔后按照Reed-Muench法分别计算两种方法在两种细胞上的TCID50。结果显示,ELISA法测定该病毒在DF-1细胞和CEF细胞上的TCID50分别为10-5.4TCID50/0.1 m L和10-4.666TCID50/0.1 m L,而IFA法测定该病毒在DF-1和CEF细胞上的TCID50分别为10-6.333TCID50/0.1 m L和10-5.2TCID50/0.1 m L。结果表明,IFA比ELISA具有更高的灵敏度,并且ALV-J在DF-1细胞上复制能力比CEF更强,在DF-1细胞上测定ALV-J的TCID50更为科学。
To compare the sensitivity and specificity between ELISA and IFA in the test of TCID50 of ALV-J,one ALV-Jstock was diluted from 10-1to 10-9and inoculated into a DF-1 cells culture in 96-well plates and CEF cells in 96-well plates re-spectively.Each dilution was inoculated into 8 wells in the same row and two cell plates were cultured with additional 7 days.Thesupernatant of each well was submitted to ELISA test for p27 antigen and the cells were detected with IFA to insure the amplifica-tion of ALV-J. The TCID50 was calculated in DF-1 and the results indicated that it is 10-5.4TCID50/0.1 m L and 10-4.666TCID50/0.1m L by ELISA and IFA respectively,while it is 10-6.333TCID50/0.1 m L and 10-5.2TCID50/0.1 m L respectively in CEF.Our study con-firmed that IFA is more sensitive than ELISA in the detection of ALV-J and the TCID50 of ALV-J identified in DF-1 is more credi-ble than in CEF.
出处
《中国兽医杂志》
CAS
北大核心
2015年第9期31-33,共3页
Chinese Journal of Veterinary Medicine
基金
公益性行业(农业)专项"种鸡场禽白血病防控与净化技术的集成"(201203055)
