LAMP PCR for Detection of Rice Tungro Virus

Abstract

Routine monitoring of tungro virus on rice field in several regions in Indonesia is necessary as an important part of disease control strategy. Therefore, a fast, accurate, and simple method to detect tungro virus is required. A research was conducted to develop loop-mediated isothermal amplification polymerase chain reaction (LAMP PCR) method as a detection approach for tungro virus. Rice leaf samples infected by tungro virus from Garut (West Java), Sidrap (South Sulawesi), and Pesisir Selatan (West Sumatera) were collected and used for this experiment. Conventional PCR was conducted as comparison to LAMP PCR. Specific DNA fragments of tungro virus (Rice tungro bacilliform virus/RTBV) was successfully amplified by conventional PCR as well as LAMP PCR. Sensitivity of LAMP PCR was higher than those of conventional PCR. Better result for detection of RTBV using LAMP PCR was achieved with sample incubation condition at 63°C for 60 min and termination reaction at 80°C for 10 min. The result of LAMP PCR can be visualized obviously using HNB dye without UV light. LAMP PCR should be recommended for routine rice tungro virus detection method due to its simplicity with accurate and high sensitivity result.