关键词: 黄瓜花叶病毒, 蔬菜, 外壳蛋白基因, 反转录环介导等温扩增技术, 检测

Abstract:

In order to develop a reverse transcription loop-mediated isothermal amplification（RT-LAMP）assay for rapid and sensitive detection of Cucumber mosaic virus（CMV）from different kind of vegetables，four primers were designed according to the coat protein（CP）gene of CMV. The reaction conditions of RT-LAMP，including the concentrations of dNTPs，Mg²⁺，reaction temperature and reaction time were optimized. The specificity and sensitivity of RT-LAMP were testified. The optimum conditions for RT-LAMP can carried out under the concentrations of 1.2 mmol · L^-1 dNTPs and 6 mmol · L^-1 Mg²⁺. RT-LAMP reaction was carried out at 61 ℃ for 40 min. The sensitivity of the RT-LAMP assay was 100-folder than that of a standard RT-PCR method. Furthermore，the products amplified by RT-LAMP could be visibly detected by the precipitate of the tube after fast centrifugation or by reacting with SYBR GreenⅠ. The RT-LAMP is highly specific，sensitive and economical，which makes it a quick method for detecting CMV from several kinds of infected vegetables and a useful technique for CMV detection in field condition.

Key words: Cucumber mosaic virus, vegetable, CP, RT-LAMP, detection