Development of a highly sensitive and rapid detection method for Pea enation mosaic virus using loop-mediated isothermal amplification assay

Abstract

The Pea enation mosaic virus (PEMV) has infected plants in the family Leguminosae such as pea, chickpea, faba bean, and lentil plants worldwide that the virus can be transmitted by sap, aphids, and seeds. Among the damages that PEMV disease cause in plants are reduced crop productivity, severely misshapen pods, wart-like outgrowths or proliferation on the surface. Previously, enzyme-linked immunosorbent assay (ELISA), reverse transcription (RT)-nested polymerase chain reaction (PCR), and real-time PCR had been used to detect PEMV. However, these methods are time-consuming and require specific equipments. For this reason, the development of a highly specific and sensitive detection method has become necessary. In this study, a new method for PEMV-1 using the loop-mediated isothermal amplification (LAMP) assay has been developed with specific primer sets as inner- and outer primers. Results showed PEMV-1 has been successfully detected that LAMP could confirm a diluted PEMV-1 up to 10−6 cDNA. LAMP is about 10,000 times more sensitive than the RT-nested PCR and/or real-time PCR. Moreover, the processing time of the LAMP was decreased 3 h than RT-nested PCR. Although future validation will be required to confirm enablement in the field area, this study provides a valuable method to identify PEMV-1 that could offer some advantages including rapid detection, high specificity and high sensitivity than others.