南方菜豆花叶病毒酶联试剂盒的测评

Evaluation of ELISA Kit for southern bean mosaic virus.

南方菜豆花叶病毒酶联检测试剂盒 ,采用直接双抗体夹心法。主要由已包被特异性抗体的酶联板、碱性磷酸酯酶标记的抗体、对硝基苯磷酸盐底物以及浓缩清洗液等几部分组成。该试剂盒的检测灵敏度可达 8ng/ml提纯病毒。检测大豆病叶可达 1 :1 0 0 0 ,检测大豆病种子可达 1 :1 0 0 ,与黄瓜花叶病毒 ,马铃薯Y病毒 ,烟草环斑病毒 ,烟草花叶病毒呈阴性反应 ,同时选用大豆等 5种健康植物作阴性对照 ,OD值读数均较低 ,背景反应好 ,未出现假阳性反应。作者对检测样品及酶标抗体的孵育时间进行摸索 ,建议样品孵育时间不宜超过 40h ,酶标抗体的孵育时间以 4h为好 ,本试剂盒的酶联板上包被有特异性抗体 ,可直接使用 ,在真空密封干燥条件下 4℃保存 ,1年内仍然有效。

Double antibody sandwich ELISA kit detecting for southern bean mosaic virus was developed and composed of specific antibody precoated plate, alkaline phosphatase labeled antibody,4 nitropheny phosphate substrate and wash buffer etc. The detection sensitivity was 8ng/ml for purified virus,1/1000 for infected soybean leaf,and 1/100 for infected soybean seed. The kit reacted negatively with cucumber mosaic virus,potato virus Y, tobacco ring spot virus and tobacco mosaic virus. the plate showed low background signal for healthy soybean plant.The author suggested that the incubation time should not be more than 40 hours for testing samples,4 hours was good for antibody AP conjugate.The kit was stable after stored at 4℃ within one year.