Development of Colorimetric Loop-Mediated Isothermal Amplification (LAMP) Technique for Rapid, Sensitive and Convenient Detection of Chrysanthemum Chlorotic Mottle Viroid (CChMVd) in Chrysanthemum

Chrysanthemum chlorotic mottle viroid (CChMVd) is a serious pathogen infecting chrysanthemum worldwide. To improve and enhance the detection procedure, colorimetric loop-mediated isothermal amplifi cation (LAMP) technique was developed. Six LAMP primers were newly designed tested optimal conditions using recombinant plasmid of CChMVd as DNA template. The for incubation at 65°C 60 min. At these conditions, ladder-liked pattern products detected along with change color from pink to yellow in positive reactions. sensitivity up 2.18 × 103 copies concentration which 104-fold greater than that polymerase chain reaction (PCR). developed not cross reacted other viruse s viroid. Effi ciency reverse transcription (RT)-LAMP detecting plantlets obtained meristem tip culture evaluated verify ensure CChMVd-free production. results showed 18 free contamination. Therefore, RT-LAMP can be used detect routinely due its accuracy, rapidness sensitivity.