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首页> 外文期刊>African Journal of Biotechnology >Detection of viruses in seeds of some vegetables by reverse transcriptase polymerase chain reaction (RT-PCR)
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Detection of viruses in seeds of some vegetables by reverse transcriptase polymerase chain reaction (RT-PCR)

机译：通过逆转录聚合酶链反应（RT-PCR）检测某些蔬菜种子中的病毒

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The present study was carried out to detect the presence of Alfalfa mosaic alfamovirus (AMV), cucumber mosaic cucumovirus (CMV), lettuce mosaic potyvirus (LMV), cucumber green mottle mosaic virus (CGMMV), tomato bushy stunt tombusvirus (TBSV), tobacco mosaic tobamovirus (TMV), tomato black ring nepovirus (TBRV) and tomato mosaic tobamovirus (ToMV) in seeds of pepper, tomato, cucumber and lettuce which are essential for human nutrition and seed production in Turkey. 50 seed samples for each vegetable obtained from various foundations and farmers were tested by reverse transcriptase polymerase chain reaction (RT-PCR) and double-antibody sandwich enzyme linked immunosorbent assay (DAS-ELISA) was conducted to compare the results. Besides, two total nucleic acid (TNA) extraction methods and a commercial RNA purification kit was compared to get high quality RNA for optimized RT-PCR. Purified total RNA extracts were used as template for cDNA synthesis, PCR products were analyzed by gel electrophoresis, and visualized by ethidium bromide staining. As a result of all, the optimum TNA extraction method was silica capture which is practical and easy to use for getting viral agents from tested vegetable seeds. RT-PCR was found rapid and sensitive in detecting viruses in seeds of vegetables when compared to DAS-ELISA. This study shows that RT-PCR can be successfully applied in certification programs and quarantine tests of vegetable seeds.
机译：进行本研究以检测是否存在苜蓿花叶苜蓿病毒（AMV），黄瓜花叶Cucumovirus（CMV），莴苣花叶potyvirus（LMV），黄瓜绿斑驳花叶病毒（CGMMV），番茄浓密特技tombusvirus（TBSV），烟草辣椒，番茄，黄瓜和生菜的种子中的花叶烟草花叶病毒（TMV），番茄黑环神经病毒（TBRV）和番茄花叶烟草病毒（ToMV），这对土耳其的人类营养和种子生产至关重要。通过逆转录酶聚合酶链反应（RT-PCR）测试了来自不同地基和农户的每种蔬菜的50个种子样品，并进行了双抗体夹心酶联免疫吸附测定（DAS-ELISA）进行比较。此外，比较了两种总核酸（TNA）提取方法和市售RNA纯化试剂盒，以获得用于优化RT-PCR的高质量RNA。纯化的总RNA提取物用作cDNA合成的模板，PCR产物通过凝胶电泳进行分析，并通过溴化乙锭染色进行可视化。总而言之，最佳的TNA提取方法是二氧化硅捕获，该方法实用且易于用于从测试的蔬菜种子中获取病毒制剂。与DAS-ELISA相比，RT-PCR被发现对检测蔬菜种子中的病毒具有快速，灵敏的作用。这项研究表明，RT-PCR可以成功地应用于蔬菜种子的认证计划和检疫测试。

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