纳米酶免疫层析试纸条法检测玉米褪绿斑驳病毒

doi:10.6048/j.issn.1001-4330.2021.06.021

新疆农业科学 ›› 2021, Vol. 58 ›› Issue (6): 1154-1159.DOI: 10.6048/j.issn.1001-4330.2021.06.021

• 园艺特产·贮藏保鲜加工·植物保护 • 上一篇下一篇

纳米酶免疫层析试纸条法检测玉米褪绿斑驳病毒

符娜¹, 向均², 张永江², 张祥林¹

1.乌鲁木齐海关技术中心,乌鲁木齐 830011;
2.中国检验检疫科学研究院,北京 100123

收稿日期:2020-05-11 出版日期:2021-06-20 发布日期:2021-06-15
通信作者: 张祥林(1964-),男,新疆乌鲁木齐人,研究员,研究方向为分子植物病理学,(E-mail)XL6479@163.com
作者简介:符娜(1994-),女,新疆昌吉人,硕士研究生,研究方向为植物检验检疫,(E-mail)1715194402@qq.com
基金资助:
国家重点研发计划(2016YFF0203200)

Study on the Detection of Maize Chlorotic Mottle Virus by Nanoenzyme Immunochromatographic Test Strip Method

FU Na¹, XIANG Jun², ZHANG Yongjiang², ZHANG Xianglin¹

1. Urumqi Customs District, Urumqi 830011,China;
2. China Academy of Inspection and Quarantine,Beijing 100123,China

Received:2020-05-11 Online:2021-06-20 Published:2021-06-15
Correspondence author: ZHANG Xianglin (1964-),Male,Urumqi,Xinjiang,Researcher,Master,the research direction: molecular plant pathology,(E-mail)XL6479@163.com

摘要/Abstract

摘要： 【目的】建立纳米酶免疫层析试纸条(Nanozyme Immuno-chromatographic Strip Assay, NISA)检测玉米褪绿斑驳病毒方法。【方法】采用双抗夹心法,将纳米酶标记的鼠抗体IgG与McmV结合后,再与硝酸纤维素膜质控线上的鼠抗体结合,二氨基联苯胺(Diaminobenzidine, DAB)滴加显色。【结果】采用研制的NISA和反转录PCR法(RT-PCR)对玉米褪绿斑驳病毒梯度稀释的研磨液进行检测,灵敏度分别为10^-7和10^-5;用其他4种病毒作对照进行特异性实验,无交叉反应;用该试纸条对供试样品进行检测,并以RT-PCR方法及基因测序方法对检测结果进行验证,三者检测结果一致。【结论】该试纸条检测McmV的灵敏度高、特异性强,可用于科研、农业生产单位和口岸机构对McmV的检测。

关键词: 玉米褪绿斑驳病毒; 纳米酶; 免疫层析试纸条

Abstract: 【Objective】 To develop a nanozyme immuno-chromatographic strip assay (NISA) to detect Maize chlorotic mottle virus. 【Method】 The double-antibody sandwich method was used to bind the nanozyme-labeled mouse antibody IgG to MCMV, and then to the mouse antibody on the nitrocellulose membrane quality control line.【Results】 The developed NISA and reverse transcription PCR (RT-PCR) were used to detect the grinding dilutions of corn chlorotic mottle virus gradient dilution, the sensitivity was 10^-7 and 10^-5 respectively; the other four viruses were used as controls for specificity. There was no cross reaction in the experiment. The test strip was used to test the test samples, and the test results were verified by RT-PCR and gene sequencing methods. The results showed that the three test results were consistent.【Conclusion】 The test strip has high sensitivity and specificity for detecting MCMV, and can be used to detect MCMV in scientific research, agricultural production units and port institutions.

Key words: Maize Chlorotic mottle virus; naozyme; immunochromatographic test paper

中图分类号:

S435.13

符娜, 向均, 张永江, 张祥林. 纳米酶免疫层析试纸条法检测玉米褪绿斑驳病毒[J]. 新疆农业科学, 2021, 58(6): 1154-1159.

FU Na, XIANG Jun, ZHANG Yongjiang, ZHANG Xianglin. Study on the Detection of Maize Chlorotic Mottle Virus by Nanoenzyme Immunochromatographic Test Strip Method[J]. Xinjiang Agricultural Sciences, 2021, 58(6): 1154-1159.

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纳米酶免疫层析试纸条法检测玉米褪绿斑驳病毒

Study on the Detection of Maize Chlorotic Mottle Virus by Nanoenzyme Immunochromatographic Test Strip Method

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