苹果茎沟病毒的提纯和检测

PURIFICATION AND DETECTION OF APPLE STEM GROOVING VIRUS

用生物学和血清学方法从苹果分离鉴定出苹果茎沟病毒G-2和TC-3分离物,采用皂土澄清、15％蔗糖垫超离心、10％～40％蔗糖梯度离心等步骤,获得提纯的病毒样品,紫外吸收比值A_(260/280)为1.15。13％SDS-PAGE法测定的病毒外壳蛋白分子量为31000。用提纯的苹果茎沟病毒G-2分离物制备的兔抗血清,PTA-ELISA测定的效价为1/64000,特异性强。用PAS-ELISA可成功地检测出苹果茎沟病毒,此外还试验了DAS-ELISA,并证明其检测苹果茎沟病毒的效果与PAS-ELISA一致。

Apple stem grooving virus (ASGV) G-2 and TC-3 were isolated from apple, and identified by both biological and serological methods. The ASGV G-2 was purified according to the procedure including bentonite clarification, PEG-6000 precipitation, 15% sucrose cushion and 10%-40% sucrose density gradient centrifugation, etc. The purified ASGV solution gave a A260/280 ratio of 1.15. The MW of coat protein of ASGV was 31000 determined by 13% SDS-PAGE. The above-mentioned virus preparation was used to immunize rabbit. The antiserum with high titer of 1 / 64000 tested by PTA-EL1SA was produced, and it displayed the strong specificity to ASGV. The PAS-ELISA has been used successfully to detect ASGV in apple. In addition, the DAS-ELISA showed the same detection efficiency of ASGV in apple.