[1]杨柳,任春梅,缪倩,等.黄瓜绿斑驳花叶病毒的双抗夹心ELISA检测[J].江苏农业学报,2018,(04):769-774.[doi:doi:10.3969/j.issn.1000-4440.2018.04.008]
　YANG Liu,REN Chun-mei,MIAO Qian,et al.DAS-ELISA detection against cucumber green mottle mosaic virus[J].,2018,(04):769-774.[doi:doi:10.3969/j.issn.1000-4440.2018.04.008]
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黄瓜绿斑驳花叶病毒的双抗夹心ELISA检测()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2018年04期
页码:
769-774
栏目:
植物保护
出版日期:
2018-08-25

文章信息/Info

Title:
DAS-ELISA detection against cucumber green mottle mosaic virus
作者:
杨柳; 任春梅; 缪倩; 陆芳; 程兆榜
（江苏省农业科学院植物保护研究所，江苏南京210014）
Author(s):
YANG Liu; REN Chun-mei; MIAO Qian; LU Fang; CHENG Zhao-bang
(Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China)
关键词:
黄瓜绿斑驳花叶病毒（CGMMV）; 单克隆抗体;  双抗夹心ELISA
Keywords:
cucumber green mottle mosaic virus (CGMMV); monoclonal antibody; DAS-ELISA
分类号:
S41-30
DOI:
doi:10.3969/j.issn.1000-4440.2018.04.008
文献标志码:
A
摘要:
为建立快速、灵敏、准确且易操作的黄瓜绿斑驳花叶病毒（CGMMV）检测方法以实现早期诊断与控制病害，制备了8株效价大于 1×10-7的单克隆抗体，配对筛选结果表明经辣根过氧化物酶标记的A8H与单抗A3配对检测的灵敏度最高，并据此创制了双抗夹心ELISA试剂盒。该试剂盒对CGMMV具有特异反应，灵敏度达 1∶12 800（g∶ml），准确度与RT-PCR结果吻合，检测周期小于5 h，可用于生产上病害发生初期的快速诊断与鉴定。
Abstract:
To establish a cucumber green mottle mosaic virus (CGMMV) detection method which was rapid, sensitive, accurate and easy to operate for diagnosing and controlling the disease in the early stage, eight hybridoma cells lines were obtained, which could secrete anti CGMMV monoclonal antibody. The titer of the monoclonal antibody was more than 1×10-7. Monoclonal antibody A3 and A8H which labeled by horseradish peroxidase were used to establish DAS-ELISA kit because of their high sensitivity. The kit had a specific response to CGMMV and the accuracy was consistent with the RT-PCR results. The detection sensitivity of leaves was 1∶12 800 (g∶ml), and the whole process was within five hours. The DAS-ELISA detection could be used for rapid diagnosis at the early stage of production.

参考文献/References:

[1]任春梅,程兆榜,缪倩,等. 江苏黄瓜绿斑驳花叶病毒的鉴定[J]. 江苏农业学报, 2013, 29(1):65-70.
[2]程兆榜,任春梅,缪倩,等. 江苏黄瓜绿斑驳花叶病毒的发生和防治[J]. 江苏农业科学, 2013, 41(2):114-117.
[3]李俊香,古勤生. 黄瓜绿斑驳花叶病毒传播方式的研究进展[J]. 中国蔬菜, 2015, 1(1):13-18.
[4]AINSWORTH G C. Mosaic disease of cucumber[J]. Annals of Applied Biology, 2008, 22(1):55-67.
[5]秦碧霞,蔡健和,刘志明,等. 侵染观赏南瓜的黄瓜绿斑驳花叶病毒的初步鉴定[J]. 植物检疫, 2005, 19(4):198-200.
[6]陈红运,赵文军,程毅,等. 辽中地区西瓜花叶病病原的分子鉴定[J]. 植物病理学报, 2006, 36(4):306-309.
[7]SLAVOKHOTOVA A A, ANDREEVA E N, SHIJAN A N, et al. Specifics of the coat protein gene in Russian strains of the cucumber green mottle mosaic virus[J]. Genetika, 2007, 43(11):1221-1226.
[8]LING K S, LI R, ZHANG W. First report of cucumber green mottle mosaic virus infecting greenhouse cucumber in Canada[J]. Plant Disease, 2014, 98(5):701-701.
[9]蔡明,李明福,江东. 日本、韩国黄瓜绿斑驳花叶病毒发生及防控策略[J]. 植物检疫, 2010, 24(4):65-68.
[10]任春梅,程兆榜,朱慧,等. 3种麦类土传花叶病毒的多抗制备及检测应用[J]. 江苏农业科学, 2014, 42(11):143-146.
[11]WU J, NI Y, LIU H, et al. Monoclonal antibody-based serological assays and immunocapture-RT-PCR for detecting Rice dwarf virus in field rice plants and leafhopper vectors[J]. Journal of Virological Methods, 2014, 195(1):134-140.
[12]SHANG H, YAN X, ZHOU X, et al. Monoclonal antibody-based serological methods for detection of cucumber green mottle mosaic virus[J]. Virology Journal, 2011, 8(1):228.
[13]陈浙,宋革,周雪平,等. 西瓜花叶病毒(WMV)单克隆抗体的制备及其应用[J]. 中国农业科学, 2016, 49(14):2711-2724.
[14]宋革,郭玉双,饶黎霞,等. 马铃薯Y病毒单克隆抗体的制备及其检测应用[J]. 浙江大学学报(农业与生命科学版), 2016, 42(5):517-526.
[15]宋西娇,刘欢,周雪平,等. BBTV单克隆抗体的制备及其检测应用[J]. 农业生物技术学报, 2016, 24(2):288-294.
[16]陈红运,赵文军,白静,等. 黄瓜绿斑驳花叶病毒cp基因的原核表达及抗血清制备[J]. 植物病理学报, 2007, 37(5):467-471.
[17]李桂芬,马洁,陈红运,等. 黄瓜绿斑驳花叶病毒多克隆抗体制备及检测应用[J]. 河南农业科学, 2007, 36(11):76-78.
[18]尚海丽,周雪平,陈青,等. 黄瓜绿斑驳花叶病毒单克隆抗体的制备[J]. 热带作物学报, 2010, 31(7):1162-1166.

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备注/Memo

备注/Memo:
收稿日期：2017-12-14 基金项目：国家自然科学基金项目（31501610）;公益性行业（农业）科研专项（201303028） 作者简介：杨柳（1984-），女，江苏徐州人，博士，助理研究员，研究方向为蔬菜病毒病。（E-mail）suiyangy@126.com 通讯作者：程兆榜，（E-mail）onlyone8501@126.com
更新日期/Last Update: 2018-09-04