›› 2013, Vol. 25 ›› Issue (6): 0-1331.

• 植物保护 •    

Rapid detection of Garlic virus X by reverse transcription loop\|mediated isothermal amplification

ZHAO Tong;LI Chang\|hong;ZHOU Qian\|jin;*;CHEN Xian\|feng;CHEN Jiong;CHEN Jian\|ping   

  1. 1 Laboratory of Biochemistry and Molecular Biology, Ningbo University, Ningbo 315211, China;2 Ningbo Academy of Science & Technology for Inspection & Quarantine, Ningbo 315012, China;3 MOA Key Laboratory for Plant Protection and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2013-11-25 Published:2013-11-25

Abstract: A one step reverse transcription loop\|mediated isothermal amplification (RT\|LAMP) assay was developed for detection of Garlic virus X(GarVX). A set of six primers that recognized eight distinct regions of GarVX were designed based on the ORF4 sequence of the GarVX. Specificity amplification was carried out with the addition of reverse transcriptase and Bst DNA polymerase. The assay was optimized, and could detect GarVX by incubation at 65℃ for only 60 min. Specificity detection showed that GarVX could be specifically detected, and no amplification was observed when the RNA template from Garlic virus A (GarVA), Garlic virus D (GarVD), or Garlic virus E(GarVE) was used. The detection limit of the RT\|LAMP method was 5 pg·μL-1, which is 10 times higher than that of RT\|PCR assay. Results demonstrated that this RT\|LAMP method can detect GarVX quickly with high specificity and sensitivity, and is suitable for field testing.

Key words: Garlic virus X, ORF4 sequence, reverse transcription loop\|mediated isothermal amplification