论文部分内容阅读
报道了应用酶联免疫测定法中的点免疫结合法 (Dot immunoblots ,Dot ELISA)和组织印迹法(Tissueblots ELISA)检测烟草组织中烟草花叶病毒 (TMV)和黄瓜花叶病毒 (CMV)的方法及效果。Dot ELISA检测提纯TMV ,兔抗TMV血清稀释 10 0 0倍 ,最低检出率可达 1.2 5ng/ml;检测TMV病叶汁液 ,最低检出稀释度为 1∶2 560。检测CMV病叶汁液 ,兔抗CMV血清稀释 50 0倍 ,最低检出稀释度为 1∶2 560。Dot ELISA检测低稀释度健康烟叶粗汁液时存在假阳性 ,提高稀释度可消除假阳性。应用Dot ELISA ,选择反应强度高、健康烟叶粗汁液无假阳性的抗血清稀释 10 0 0倍 (TMV)、50 0倍 (CMV) ,样品粗汁液稀释30 0倍 ,以及应用组织印迹法对采自云南曲靖市的花叶样品进行了TMV和CMV检测 ,并与电镜负染法的检测结果验证。表明两种方法都可以用来检测烟叶样品。组织印迹法比Dot ELISA法的操作简单 ,灵敏度高 ,适宜大量样品的常规检测
Reported the detection of tobacco mosaic virus (TMV) and cucumber mosaic virus (CMV) in tobacco tissues by dot immunoblots (Dot ELISA) and Tissueblots ELISA in ELISA And the effect. Dot ELISA detection of purified TMV, rabbit anti-TMV serum diluted 100 times, the lowest detection rate of up to 1.2 5ng / ml; TMV disease leaf juice detection, the lowest detected dilution of 1: 560. CMV diseased leaf juice was tested. Rabbit anti-CMV serum was diluted 50 times and the lowest detected dilution was 1: 560. Dot ELISA detects false-positives in low-dilution healthy juices, and increases the dilution to eliminate false-positives. Dot ELISA was used to select TMV, 50 0 times (CMV) and 300 times dilution of crude juice of healthy tobacco leaves with no false positive, TMV and CMV were detected from the mosaic samples of Qujing City, Yunnan Province, and the results were verified by electron microscopy negative staining. This shows that both methods can be used to detect tobacco samples. Tissue blotting method is simpler and more sensitive than Dot ELISA method, suitable for routine testing of a large number of samples