生物技术通报 ›› 2019, Vol. 35 ›› Issue (12): 184-188.doi: 10.13560/j.cnki.biotech.bull.1985.2019-0287

• 技术与方法 • 上一篇    下一篇

马铃薯S病毒胶体金免疫层析试纸条的研制

张微, 李志新, 付春江, 刘卫平   

  1. 黑龙江省农业科学院克山分院,齐齐哈尔 161600
  • 收稿日期:2019-04-08 出版日期:2019-12-26 发布日期:2019-12-03
  • 作者简介:张微,硕士,研究方向:马铃薯病毒检测技术与马铃薯病害防控;E-mail:zw13936823026@126.com
  • 基金资助:
    黑龙江省现代农业马铃薯产业技术协同创新体系项目(HNWJZTX201701)

Development of a Colloidal Gold Immunochromatographic Test Strip for the Detection of Potato Virus S

ZHANG Wei, LI Zhi-xin, FU Chun-jiang, LIU Wei-ping   

  1. Keshan Branch of Heilongjiang Academy of Agricultural Sciences,Qiqihar 161600
  • Received:2019-04-08 Published:2019-12-26 Online:2019-12-03

摘要: 为了研制便于马铃薯(Solanum tuberosum L.)种薯生产田现场快速检测马铃薯S病毒(Potato virus S,PVS)的胶体金免疫层析试纸条。采用柠檬酸三钠还原法制备胶体金,在波长524 nm处有最大吸收值,金颗粒大小20-30 nm。将PVS的羊源多克隆抗体与胶体金进行标记,喷射于玻璃纤维上作为金标垫。将PVS抗体和兔抗羊抗体划线于硝酸纤维素膜上作为检测线和质控线,制备胶体金试纸条。试纸条检测结果在2 min之内通过肉眼即可判断。当带PVS样本汁液稀释10 4倍(W/V)时,仍可检测出。用其他5种常见病毒阳性样品进行检测,未出现交叉反应。田间采集的马铃薯叶片用试纸条检测和酶联免疫检测结果相吻合。本试纸条在密封、干燥、低温的条件下保存,有效期可达180 d。使用方便,操作简单,检测快捷,能够现场检测PVS,无需特殊设备,适用性广,实用性强。

关键词: 马铃薯S病毒, 胶体金免疫层析法, 快速检测, 试纸条

Abstract: This work aims to develop a colloidal gold immunochromatographic test strip for on-site rapid detection of potato(Solanum tuberosum L.)virus S(PVS). Colloidal gold solution was prepared by trisodium citrate reduction method,its maximum absorption value was at the wavelength of 524 nm and the particle size was 20-30 nm. In the strip test,colloidal gold-labeled polyclonal sheep antibody against PVS was sprayed on the glass fiber as the detection antibody,and rabbit anti-goat IgG at the control line and anti-PVS at the test line on the nitrocellulose membrane of the test strip was served as the capture antibody. The positive result was easily judged by the presence of a red test line with naked eye within 2 min. The detection was still feasible when the samples was diluted by 104 W/V,and test strip showed no cross effect when tested by other common 5 viruses(PVX,PVY,PLRV,PVA,and PVM)samples. The detection of potato leaves from field by the test strip was consistent with that by ELISA. Test strip is valid for 180 d while stored in dry sealed low-temperature 4℃. The test strip is convenient,simple and rapid in the field detection of PVS,no additional equipment is required,thus applicability and practicability are wide.

Key words: potato virus S, colloidal gold immunochromatography assay, rapid detection, test strip