WEI Mei-sheng,YOU Jia,MA Jie,et al.Colloidal Gold Immunochromatographic Strip for Rapid Detection of Lettuce mosaic virus[J].HEILONGJIANG AGRICULTURAL SCIENCES,2014,(11):74-77.
莴苣花叶病毒胶体金免疫层析试纸条的研制
- Title:
- Colloidal Gold Immunochromatographic Strip for Rapid Detection of Lettuce mosaic virus
- 文章编号:
- 1002-2767(2014)11-0074-04
- 分类号:
- S432.4+1
- 文献标志码:
- A
- 摘要:
- 为建立莴苣花叶病毒的现场快速检测方法,采用胶体金标记莴苣花叶病毒的兔多克隆抗体,在硝酸纤维素膜上喷涂检测T线和质控C线,制作胶体金免疫层析试纸条。该试纸条检测莴苣花叶病毒提纯病毒的灵敏度为1 μg?mL-1,检测昆诺阿藜病汁液可稀释1 000倍(重量/体积)。试纸条可特异性检测到莴苣花叶病毒的3个分离物,与马铃薯X等8种病毒无交叉反应。田间采集的莴苣样品试纸条和酶联检测的结果一致。该试纸条适用于疑似莴苣花叶病毒感染的莴苣叶片的快速诊断,并在10 min内获得检测结果。
- Abstract:
-
In order to establish a rapidly diagnostic method for Lettuce mosaic virus(LMV)in the field,collioidal gold immunochromatographic strip(ICS)was developed by coupling colloidal golds with polyclonal rabbit antibody against LMV.Test line and control line on nitrocellulose membrane were spread by Biojet Quanti 3 000 dispenser on XYZ dispensing platform.The sensitivity of immunostrip was 1 μg·mL-1 for purified LMV.Extract from LMV infected Chenopodium quinoa leaf diluted 1 000 times could be used to detect(W/V).The immunostrip was specific to LMV and detected three isolated viruses.There was no cross reaction with other eight viruses.Immunostrip test result was confirmed by ELISA for lettuce leaves collecting from field.The immunostrip was suitable for rapid diagnosis of LMV in suspicious lettuce leaf samples within 10 minutes.
参考文献/References:
[1]Le Gall O.Lettuce mosaic virus.Descriptions of plant viruses[EB/OL].[2014-06-01]]http://www.dpvweb.net/dpv/,2003:No.399.
[2]夏俊强.国外莴苣花叶病毒的研究[J].莱阳农学院学报,1985(1):133-138.
[3]Naidu,R A,Karthikeyan G,Jarugula S,et al.First report of the natural infection of Coreopsis auriculata‘Nana’ with Lettuce mosaic virus in the United States[J].Plant Disease,2008,92(3):486.
[4]夏俊强,严敦余,王清和,等.莴苣花叶病毒的研究Ⅰ.病原鉴定[J].植物病理学报,1984,14(4):241-246.
[5]魏宁生,贾长盛.莴苣病毒病病毒的分离及鉴定[J].病毒学报,1987,3(1):102-103.
[6]钱荣田,骆江洪,龚志强.侵染莴苣的莴苣花叶病毒(LMV)研究[J].浙江农业大学学报,1996,22(3):304-307.
[7]夏俊强,严敦余,王清和,等.莴苣花叶病毒病的研究Ⅱ病害的分布、损失、寄主和传播[J].植物病理学报,1986,16(1):37-40.
[8]Revers F,Lot H,Souche S,et al.Biological and molecular variability of lettuce mosaic virus isolates[J].Phytopathology,1997,87:397-403.
[9]郑滔,陈炯,陈剑平.莴苣花叶病毒浙江余杭分离物基因组全序列及其结构分析[J].病毒学报,2002,18(1):66-70.
[10]尚巧霞,向海英,韩成贵,等.莴苣花叶病毒北京分离物基因组3`末端序列分析[J].农业生物技术科学,2007,23(7):88-90.
[11]Falk,B W,Purcifull,D E.Development and application of an enzymelinked immunosorbent assay(ELISA)test to index lettuce seeds for lettuce mosaic virus in Florida[J].Plant Disease,1983,67:413-416.
[12]Peypelut M,KrauseSakate R,Guiraud T,et al.Specific detection of lettuce mosaic virus isolates belonging to the “Most” type\[J\].Journal of Virological Methods,2004,121(1):119-24. [13]周雪平,濮祖芹.侵染豌豆的莴苣花叶病毒(LMV)研究[J].江苏农业学报,1991,7(2):4447.
[14]魏梅生,刘洪义,李桂芬,等.马铃薯X病毒和马铃薯Y病毒胶体金免疫层析试纸条的研制[J].植物保护,2006,32(6):139-141.
备注/Memo
收稿日期:2014-06-10
基金项目:质检公益性行业科研专项资助项目(201310071)
第一作者简介:魏梅生(1965-),男,安徽省巢湖市人,硕士,研究员,从事植物病毒病害的检疫检测研究。E-mail:xsms2013@163.com。