In the present study, two isothermal molecular assays viz. reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) and reverse transcriptase recombinase amplification (RT-RPA) were developed to detect the cardamom vein clearing virus (CdVCV) infecting cardamom. Assays were optimized for parameters like duration, temperature and concentration of magnesium sulfate, and betaine in the case of RT-LAMP and magnesium acetate in the case of RT-RPA. Detection limits of both assays were determined and compared with conventional RT-PCR and SYBR Green-based real-time RT-PCR. RT-LAMP was found 10,000 times additional sensitive than RT-PCR and one-tenth that of real-time RT-PCR. RT-RPA was found 1000 times additional sensitive than RT-PCR and one-hundredth that of real-time RT-PCR. Both assays were specific, rapid, and sensitive for detecting CdVCV. Compared to real-time RT-PCR, these assays are economical and can be employed in large scale screening of cardamom plants against CdVCV for the selection of virus-free plants.

中文翻译：

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逆转录酶环介导的等温扩增和逆转录酶重组酶扩增测定法可快速灵敏地检测豆蔻静脉清洁病毒。

在本研究中，两个等温分子测定法。开发了逆转录酶环介导的等温扩增（RT-LAMP）和逆转录酶重组酶扩增（RT-RPA），以检测感染豆蔻的豆蔻静脉清除病毒（CdVCV）。对于时间，温度和浓度（如硫酸镁和甜菜碱）（如使用RT-LAMP）和醋酸镁（如使用RT-RPA），对测定进行了优化。确定了两种测定的检出限，并与常规RT-PCR和基于SYBR Green的实时RT-PCR进行了比较。发现RT-LAMP的灵敏度是RT-PCR的10,000倍，是实时RT-PCR的十分之一。发现RT-RPA的敏感性是RT-PCR的1000倍，是实时RT-PCR的一百倍。两种检测都是特异性，快速，对检测CdVCV敏感。与实时RT-PCR相比，这些测定是经济的，可用于针对CdVCV的豆蔻植物的大规模筛选，以选择无病毒的植物。