HUO Yayun,ZHANG Yongjiang,LI Weimin,et al.Development of Nucleic Acid Test Strip for Detection of Cucumber Green Mottle Mosaic Virus[J].Northern Horticulture,2017,41(10):100-103.[doi:10.11937/bfyy.201710023]
黄瓜绿斑驳花叶病毒核酸试纸条检测技术的建立
- Title:
- Development of Nucleic Acid Test Strip for Detection of Cucumber Green Mottle Mosaic Virus
- 文献标志码:
- A
- 摘要:
- 以黄瓜绿斑驳花叶病毒（cucumber green mottle mosaic virus，CGMMV）阳性样品为研究对象，根据其外壳蛋白基因设计3条引物和2条探针，探针5′端分别用生物素和荧光进行标记；利用这套引物和探针在59 ℃进行RT-PCR恒温扩增90 min，扩增产物用核酸试纸条进行检测。用其它5种同样侵染葫芦科作物的植物病毒作为对照进行试验，以期确定试纸条检测方法的特异性。将梯度稀释后的总RNA作为模板进行检测，以确定该方法的灵敏度。结果表明：该试验建立的核酸试纸条检测方法对CGMMV具有特异性，能够有效区分CGMMV及同样侵染葫芦科作物的黄瓜花叶病毒（CMV）、辣椒轻斑驳病毒（PMMoV）、烟草花叶病毒（TMV）、番茄花叶病毒（ToMV）和小西葫芦黄花叶病毒（ZYMV）；同时该方法也具有较高的灵敏度，可检测低至24.300 ng?μL-1的总RNA。因此该试验建立的核酸试纸条法可用于CGMMV的快速检测。
- Abstract:
- Positive sample of CGMMV was used as research object.Three primers and two probes were designed according to the coat protein of CGMMV and probes were labeled with Biotin and FAM at 5′ terminus separately.The nucleic acid strip was used to show the detection result after isothermal RT-PCR at the temperature of 59 ℃ for 90 minutes.The other five viruses that also infect cucurbitaceae crops were used to assay the specificity of this method.Diluted nucleic acid solution with certain concentration was used as template to confirm the sensitivity of this method.The results showed that nucleic acid strip was a special method to distinguish CGMMV from the other five plant virus which also infect cucurbitaceae crops，CMV，PMMoV，TMV，ToMV and ZYMV.And the sensibility of this method was 24.300 ng?μL-1 of total RNA.Thus，nucleic acid strip was a rapid，special and sensitive detection method for CGMMV and can be used in routine testing.
参考文献/References:
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相似文献/References:
[1]宋顺华,吴 萍,宫国义,等.黄瓜绿斑驳花叶病的发生及防控策略[J].北方园艺,2014,38(08):105.
SONG Shun-hua,WU Ping,GONG Guo-yi,et al.Occurrence and Control Measures of Cucumber Green Mottle Mosaic Virus[J].Northern Horticulture,2014,38(10):105.
备注/Memo
第一作者简介：霍亚云（1993-），女，硕士研究生，研究方向为生物化学与分子生物学。E-mail：huoyayuns@163.com.责任作者：张永江（1977-），男，博士，研究员，现主要从事植物检疫等研究工作。E-mail：zhangyjpvi@yeah.net.基金项目：国家“十二五”科技支撑计划资助项目（2015BAD08B01）；国家“十三五”重点研发计划资助项目（2016YFF0203203）。