INTERNATIONAL JOURNAL OF AGRICULTURAL TECHNOLOGY

Volume 15, No. 02, Month MARCH, Year 2019, Pages 215 - 228

Enhancing columnea latent viroid detection using reverse transcription loop-mediated isothermal amplification (rtlamp)

Bhuvitarkorn, S., Klinkong, S. and Reanwarakorn, K.

Abstract Download PDF

Columnea latent viroid (CLVd) is an important quarantine plant pathogen, it is of regulatory interest because of its high rate of seed transmission in tomato (Solanum lycopersicum). The detection of CLVd from leaves and seeds of solanaceous and cucurbitaceae species were investigated. A reverse transcription loop-mediated isothermal amplification (RTLAMP) method was developed for detection of CLVd within 1 h. The method is highly sensitive, being 2,000 times and 100 times more sensitive than conventional RT-PCR in detecting CLVd from leaves and seeds, respectively. The results showed that this detection method was species specific to CLVd with no cross reactivity to other viroids in the same Pospiviroid genus. It was successfully applied to detect CLVd from field samples. This appears to be the first application of RT-LAMP for detecting CLVd.

Keywords

Detection, RT-LAMP, viroids, leaves, seeds

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