多重烟草病毒胶体金检测试纸条的研制及应用

Preparation and application of colloidal gold test paper for the detection of multiple tobacco viruses

摘要: 为建立一种快速、特异、简便的多重烟草病毒检测方法，用胶体金标记和免疫层析技术研制了双重（TMV-CMV、CMV-PVY、TMV-PVY）和三重（TMV-CMV-PVY）病毒试纸条。测试结果表明，三重病毒检测试纸条对目标病毒（TMV、CMV和PVY）的检测具有良好的灵敏性和特异性。检出TMV、CMV和PVY的最低浓度分别在1.02~10.2、1.10~11.0、1.20~12.0 ng/mL之间。对烟草危害严重的TEV、PVA和TSWV均无交叉反应。用三重病毒检测试纸条对苗棚中的烟株进行检测，其结果与RT-PCR检测结果的符合率≥90%。

Abstract: To establish a rapid, specific and simple method for detecting tobacco virus, a colloidal gold labeling technique and immunochromatography were incorporated into paper-based test strips to target two (TMV-CMV, CMV-PVY and TMV-PVY) or three (TMV-CMV-PVY) viruses. The results showed that the test paper for the detection of three viruses had good sensitivity and specificity. The minimum detectable concentrations for TMV, CMV and PVY were 1.02-10.2, 1.10-11.0 and 1.20-12.0 ng/mL, respectively. No cross reaction between TMV, CMV, PVY and TEV, PVA, TSWV were found with the test paper for these harmful tobacco viruses. Tobacco seedlings in greenhouses were also subjected with the test paper, and the coincidence rate was above 90% between the detected results using the test paper and a RT-PCR technique.