Subscribe Now Subscribe Today
Science Alert
 
Blue
   
Curve Top
Plant Pathology Journal
  Year: 2012 | Volume: 11 | Issue: 2 | Page No.: 51-59
DOI: 10.3923/ppj.2012.51.59
 
Facebook Twitter Digg Reddit Linkedin StumbleUpon E-mail

Detection of Kyuri Green Mottle Mosaic Virus from Soil by the Immunocapture Reverse Transcription Loop-mediated Isothermal Amplification Reaction

S. Fukuta, K. Takeyama, M. Suzuki, A. Shichi, K. Ichikawa and H. Nakanishi

Abstract:
Kyuri green mottle mosaic virus (KGMMV) which is a major soil-borne virus, causes severe yield reduction in cucumber in Japan. We developed immunocapture reverse transcription loop-mediated isothermal amplification (Ic/RT-LAMP) method to detect KGMMV in soil. Degenerate primer set from KGMMV C strain (KGMMV-C) and Yodo strain (KGMMV-Yodo) was designed for the detection of both strains. To optimize the reaction condition, the RT-LAMP reaction using the degenerate primer set was performed at different temperature (60-65°C). The reaction at 60°C gave the best results. The result on specificity test using 7 cucumber pathogenic viral isolates indicated that the RT-LAMP assay established in this study had no cross reactions. The detection limit of the RT-LAMP assay was 10 pg of purified KGMMV. The sensitivity of RT-LAMP was 10 times higher than that of RT-PCR. Detection limit of DAS-ELISA and Ic/RT-LAMP was compared by using of the dilution of purified KGMMV-C in soil. Ic/RT-LAMP could detect 0.5 ng of KGMMV from 100 mg of soil sample and the sensitivity of Ic/RT-LAMP was 200 times higher than that of DAS-ELISA. The Ic/RT-LAMP assay could successfully detect KGMMV in soil samples from 10 fields where KGMMV had been detected in a previous cultivation. The present Ic/RT-LAMP method showed to be a rapid, simple and available detection method of KGMMV in soil.
PDF Fulltext XML References Citation Report Citation
 RELATED ARTICLES:
  •    Double Infections with Cucumber Mosaic Virus and Plum Pox Virus (Sharka) in Apricot Trees
  •    Biological, Serological and Molecular Diagnosis of Three Major Potato Viruses in Egypt
  •    Efficacy of Mulches and Methyl Bromide Alternatives on Soilborne Pests and Weeds in Spring Tomato
  •    Molecular Detection and Partial Characterization of Begomovirus Associated with Leaf Curl Disease of Sunflower (Helianthus annuus) in Southern India
How to cite this article:

S. Fukuta, K. Takeyama, M. Suzuki, A. Shichi, K. Ichikawa and H. Nakanishi, 2012. Detection of Kyuri Green Mottle Mosaic Virus from Soil by the Immunocapture Reverse Transcription Loop-mediated Isothermal Amplification Reaction. Plant Pathology Journal, 11: 51-59.

DOI: 10.3923/ppj.2012.51.59

URL: https://scialert.net/abstract/?doi=ppj.2012.51.59

COMMENT ON THIS PAPER
 
 
 

 

 
 
 
 
 
 
 
 
 

 
 
 
 
 

Curve Bottom