一种快速简便检测马铃薯病毒的方法——病毒细菌协同凝集法(VBA)的研究

 在国内、外首次选用金黄色葡萄球菌(Staphylococc aureus)的No.180菌株用于病毒细菌协同凝集试验(Virobacterial agglutination test简称VBA)检测了来自马铃薯的4种不同形态粒子的病毒:马铃薯X病毒(PVX)、马铃薯丫病毒(PVY)、烟草花叶病毒(TMV)和马铃薯卷叶病毒(PLRV),其灵敏度达2.7~6.1ng/ml,检出病汁液的最大稀释度达10⁴~10⁵(PLRV1:500),较国内、外采用的Cown I菌株的灵敏度提高5~10倍。与血清学方法相比,VBA在2~3分种内就可获得结果,无假阳性反应,其灵敏度显著高于间接酶联法和免疫电镜,而接近于A蛋白酶联法。经抗血清致敏的菌体在4℃下保存4个月,对VBA检测的灵敏度没有影响。用VBA对采自田间的93个马铃薯病样进行检测,它们大多受2~3种病毒(PVX、PVY及PLRV)的复合侵染;和用间接酶联法及鉴别寄主检测的结果趋向一致。室内和田间试验均表明VBA灵敏度高、特异性强、快速简便和经济,尤其适合在基层单位中推广应用。

VIROBACTERIAL AGGLUTINATION TEST(VBA)——A SIMPLE AND RAPID METHOD FOR DETECTING POTATO VIRUSES

Strain No. 1800 of Staphylococcus aureus was first used in Virobacterial agglutination test (VBA) to have detected four types of morphological potato viruses (PVX,PVY,TMV and PLRV). The sensitivities are 5-10 times higher than those from using strain Cown I and Walkey modified method of bacteria cell suspension prepartion, which can be reached 2.7-6.1ng/ml for purified viruses and 10⁴~10⁵ for infected saps except PLRV (1:500). The storage period of cell suspension was not effected on the sensitivities of the VBA and which were to be the highest among IM-ELISA.IEM,MPT,CIEP and IDD, almost equal to SPA-ELISA. Also, our results have proved that the specificity of the VBA was more high than those of the others. By using VBA and IM-ELISA, 93 infected potato samples form fields were detected and most of the samples were infected by 2-3 viruses (PVX, PVY and PLRV). The results from the indicator plant method were consistent with those from VBA and IM-ELISA. All of the results have proved that the VBA is a sensitive, specific, rapid, economic, simple and applicable method for detecting viruses of potato or of other plants, especially for the use in basic units.