烟草花叶病毒胶体金检测试纸条的制备与性能评价

王春琼; 刘凯; 谢永辉; 张轲; 刘春明; 张晓伟; 孙浩巍; 陈丹

doi:10.12187/2025.01.011

February 2025

Article Contents

WANG Chunqiong, LIU Kai, XIE Yonghui, et al. Preparation and performance evaluation of colloidal gold test strips for tobacco mosaic virus detection[J]. Journal of Light Industry, 2025, 40(1): 90-97. doi: 10.12187/2025.01.011

Citation: WANG Chunqiong, LIU Kai, XIE Yonghui, et al. Preparation and performance evaluation of colloidal gold test strips for tobacco mosaic virus detection[J]. Journal of Light Industry, 2025, 40(1): 90-97. doi: 10.12187/2025.01.011 shu

Preparation and performance evaluation of colloidal gold test strips for tobacco mosaic virus detection

1. Yunnan Tobacco Quality Supervision and Test Station, Kunming 650106, China;
2. Yunnan Tobacco Company Kunming City Company, Kunming 650000, China;
3. Yunnan Tobacco Company Honghe Prefecture, Mile 652300, China

Corresponding author: CHEN Dan, chen.dana@qq.com
Received Date: 2024-01-23
Accepted Date: 2024-03-21

Abstract

To address the problems of the time-consuming and limited application of traditional tobacco mosaic virus (TMV) detection methods, colloidal gold test strips for rapid and qualitative detection of TMV were developed based on the double antibody sandwich method, and its preparation conditions were optimized.In addition, the limit, accuracy, and specificity of the test strip were detected. The results showed that the suitable preparation conditions for the test strip were buffer addition of 4 μL, antibody mass concentration of 10 μg/mL, and NC membrane of CN140 type and sample extraction solution of 0.02 mol/L PBS; The detection limit of this test strip for TMV-CP protein was 0.1 μg/mL, with no cross reactivity with tobacco bush top virus, tomato spot wilt virus, and potato Y virus, and good specificity; Applied to actual sample detection, the total coincidence rate with PCR was 95.0 %, the positive coincidence rate was 93.3 %, and the accuracy was good. This test strip could display the detection results within 10 min and was easy to operate. It could be used for rapid detection of TMVs in tobacco field.
- tobacco mosaic virus,
- monoclonal antibody,
- colloidal gold immunochromatographic technique,
- test strips
References
1. [1]
  吴彦辉,蒲团卫,张生杰,等.烟草花叶病毒实时荧光定量PCR检测体系的构建[J].安徽农业科学,2023,51(12):171-175.
2. [2]
  付一峰.烟草花叶病毒病的研究进展[J].北京农业,2015(12):115-117.
3. [3]
  刘羽丰,徐晨曦,吴泽,等.北京市昌平地区园林植物烟草花叶病毒的检测[J].北京农学院学报,2022,37(1):28-31.
4. [4]
  台莲梅,靳学慧,张亚玲,等.黑龙江烟草病害种类鉴定及发生情况[J].贵州农业科学,2014,42(10):124-126.
5. [5]
  姜瀚林,郭兆奎,刘永中,等.3个黑龙江烟区烟草花叶病毒分离物的全基因组序列测定与分析[J].中国烟草学报,2019,25(1):77-85.
6. [6]
  白静科,何雷,吴彦辉,等.河南烟草主要病毒发生种类及侵染类型分析[J].中国烟草科学,2022,43(1):49-54.
7. [7]
  万晴姣,张晓霞,李欲轲,等.贵州辣椒烟草花叶病毒的分子鉴定[J].贵州农业科学,2015,43(10):181-184.
8. [8]
  蹇军友,刘开兴,耿召良,等.黔产乌头中的生物碱成分及其抗烟草花叶病毒(TMV)活性[J].农药,2017,56(10):767-770
  ,777.
9. [9]
  李丽珠.云南烟草主要病虫害分类及防治技术[J].热带农业工程,2019,43(2):14-16.
10. [10]
  KUMAR S,PRAKASH H S.Detection of tobacco mosaic virus and tomato mosaic virus in pepper seeds by enzyme linked immunosorbent assay (ELISA)[J].Archives of Phytopathology and Plant Protection,2016,49(1/2/3/4):59-63.
11. [11]
  XU T Z,LIN X W,ZHANG X L,et al.Triplex visual detection of tobacco potyviruses using reverse transcription recombinase polymerase amplification assay combined with lateral flow dipstick[J].Crop Protection,2023,174:106397.
12. [12]
  LI Y Y,TAN G L,LAN P X,et al.Detection of tobamoviruses by RT-PCR using a novel pair of degenerate primers[J].Journal of Virological Methods,2018,259:122-128.
13. [13]
  任锡毅,刘永翔,黄永会,等.试管捕捉实时荧光RT-PCR法同时检测烟草花叶病毒病和马铃薯Y病毒病[J].江苏农业科学,2017,45(6):96-100.
14. [14]
  董旭旭,孙威,曹攀,等.胶体金免疫层析试纸条技术在病毒检测领域的应用研究现状[J].生物工程学报,2022,38(9):3243-3254.
15. [15]
  赵少若,黄甜,郝丽影,等.犬轮状病毒单克隆抗体制备及胶体金试纸条检测方法的建立[J].中国畜牧兽医,2022,49(11):4429-4437.
16. [16]
  SHYAM K U,JEONG H N,OH M J,et al.Development of a lateral flow immuno-chromatic strip assay for the detection of nervous necrosis virus (NNV,RGNNV genotype)[J].Aquaculture,2020,520:734944.
17. [17]
  刘国华,蒋永青,陈彤,等.安眠酮胶体金免疫快速检测试纸条的研制[J].国外畜牧学(猪与禽),2023,43(6):64-70.
18. [18]
  岳绪辉,杜斌,林栋,等.草甘膦胶体金免疫层析试纸条的研制[J].食品科技,2021,46(12):301-307.
19. [19]
  韩金治,李亚茹,罗林,等.胶体金免疫层析法快速检测水产品中河豚毒素的研究[J].分析测试学报,2023,42(12):1598-1606.
20. [20]
  姜宁,夏振远,卢灿华,等.烟草番茄环纹斑点病毒快速检测试纸条的研制及应用[J].烟草科技,2023,56(11):10-15
  ,53.
21. [21]
  杨洋,羊国根,杨懿德,等.烟草黑胫病菌胶体金免疫层析快速检测技术的建立与评价[J].安徽农业科学,2022,50(17):128-131
  ,137.
22. [22]
  周立娟,井申荣,苑荣亮,等.夹心ELISA检测新冠病毒及其抗原方法的建立和验证[J].云南大学学报(自然科学版),2022,44(4):870-876.
23. [23]
  刘云翔,高鹏超,詹志来,等.基于酶联免疫分析的中药薏苡仁玉米赤霉烯酮残留快速检测技术研究[J].中国中药杂志,2023,48(11):2919-2924.
24. [24]
  张文劲,李庆妮,唐定,等.恒河猴血清SARS-CoV-2变异株特异性IgG抗体间接ELISA检测方法的建立及验证[J].中国生物制品学杂志,2023,36(11):1341-1346
  ,1352.
25. [25]
  KUANG H,XING C R,HAO C L,et al.Rapid and highly sensitive detection of lead ions in drinking water based on a strip immunosensor[J].Sensors,2013,13(4):4214-4224.
26. [26]
  曹涤非,薛佳莹,崔向红,等.柠檬酸三钠法制备胶体金及其应用[J].化学工程师,2019,33(12):12-14
  ,24.
27. [27]
  刘靖.非洲猪瘟病毒CD2v、p30和pK205R蛋白的表达纯化与p30、pK205R蛋白单克隆抗体的制备[D].北京:中国农业科学院,2021.
28. [28]
  侯浩宇,郑南南,吴宏举,等.非洲猪瘟病毒p30蛋白单克隆抗体制备和鉴定[J].中国畜牧兽医,2023,50(3):1241-1249.
29. [29]
  王志超,马秋敏,叶彦雄,等.番茄Ⅱ型metacaspase蛋白SlMC7抗体效价检测及其在果实中表达特性分析[J].北方园艺,2017(16):50-55.
30. [30]
  孔卫青,凌君,杨金宏.桑花叶卷叶病相关病毒外壳蛋白抗体的制备及利用[J].西北农业学报,2021,30(7):1077-1082.
31. [31]
  HU J,ZHOU S,ZENG L F,et al.Hydrazide mediated oriented coupling of antibodies on quantum dot beads for enhancing detection performance of immunochromatogr-aphic assay[J].Talanta,2021,223(Pt 1):121723.
32. [32]
  王康,余璞,陈孔,等.应用胶体金免疫层析法检测饲料中恩拉霉素残留的方法建立[J].生物工程学报,2024,40(7):2346-2356.
33. [33]
  TANG R H,ALAM N,LI M,et al.Dissolvable sugar barriers to enhance the sensitivity of nitrocellulose membrane lateral flow assay for COVID-19 nucleic acid[J].Carbohydrate Polymers,2021,268:118259.
34. [34]
  柯童炜,骆骄阳,秦家安,等.胶体金免疫层析法快速检测中药材中除草剂扑草净残留[J].药物分析杂志,2024,44(1):174-184.
35. [35]
  刘欣.烟草花叶病毒胶体金检测试纸条的研制[D].郑州:河南农业大学,2009.
36. [36]
  张敏,李建,舒金秀,等.犬瘟热病毒单克隆抗体的研制及胶体金试纸条检测方法的建立[J].中国兽药杂志,2022,56(12):16-24.
37. [37]
  吕琦,吴峰,林洁,等.烟草花叶病毒单克隆抗体的制备和鉴定[J].云南大学学报(自然科学版),2013,35(1):93-98.
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Preparation and performance evaluation of colloidal gold test strips for tobacco mosaic virus detection

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