应用A蛋白酶联免疫吸附法检测烟草花叶病毒

Detection of Tabacco Mosaic Virus (TMV) with Stephylococal Protein A-ELISA:

A蛋白酶联免疫吸附分析间接法(ELISA-SPA)被成功地用于烟草花叶病毒(TMV)的检测。本方法两次将A蛋白用于抗体-抗原-抗体双夹心层。第1次是将A蛋白包被于反应板,加双抗体夹心后,第2次将酶联A蛋白包被抗体中IgG的Fc段,阻止了非特异性反应。本方法检测TMV的灵敏度:提纯抗原为0.5ng/ml,病叶澄清液为稀释10~8。此法比双抗体夹心法(ELISA-DAS)和异种动物双抗体夹心法(ELISA-DSM)均高,并在8小时内就可获得结果,适用于大量样品的检测。

A nevel form of indirect enzyme-linked immunosorbent assay (ELISA-SPA) has been used for the detection of TMV in purified preparation and infected tomato sap. With two application of protein A to be sandwiched between antibody-antigen-antibody layers, the method was done into two steps. The first step is to attach protein A on the reacting plate as solid coating layer; after adding double antibody, the following step is to use the conjugated enzyme with protein A to coat Fc segment of IgG immunoglobulin preventing the nonspecific reaction. The sensitivity of the method for detecting TMV is that the antigen should be purified up to 0.5ng/ml and the dilution of infected sap to 10~8, It is showed thay the method is more sensitive than ELISA-DAS and ELISA-DSM, which requires only 8 hours to complete the operation and fits to detect the large sample.