双重检测马铃薯X和Y病毒试纸条制备技术研究
Crops（2021）
Abstract
为了实现在马铃薯种薯生产田现场同时快速检测马铃薯X (PVX)和Y病毒(PVY),利用胶体金标记和免疫层析技术研制了PVX-PVY双重检测试纸条.采用柠檬酸三钠还原法制备胶体金溶液,在波长526nm处有最大吸收值,金颗粒直径约25nm.同时标记PVX和PVY兔多克隆抗体,将抗体标记物喷射于玻璃纤维纸上,作为金标垫.将PVX和PVY抗体分别划线于硝酸纤维素膜上作为检测线,将羊抗兔抗体划线于硝酸纤维素膜上作为质控线,制备胶体金试纸条.结果 表明,研制的双重试纸条能够在2min之内同时检出PVX和PVY.PVX病汁液检测稀释限度可达106倍(W/V),PVY可达105倍(W/V),其中对PVX检测更灵敏.用其他4种常见病毒(PVS、PLRV、PVA和PVM)检测,未出现交叉反应.在田间采集马铃薯叶片,分别利用试纸条与DAS-ELISA检测,结果一致性非常高.由此可知,该试纸条具有操作简便、反应快捷的特点,特别适用于田间及口岸一线对于马铃薯X和Y病毒的同时检测.
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