Development of transcription recombinase polymerase based isothermal amplification coupled with lateral flow immunochromatographic assay for visual detection of citrus tatter leaf virus

J Virol Methods. 2022 Nov:309:114593. doi: 10.1016/j.jviromet.2022.114593. Epub 2022 Jul 28.

Abstract

The citrus tatter leaf virus (CTLV) is one of the most destructive citrus viral diseases worldwide. In this study, reverse transcription-recombinase polymerase amplification combined with a lateral flow dipstick (RT-RPA-LFD) assay for rapid visual detection of CTLV was established. The assay was performed at 35 ℃ in 27 min without specialised equipment. The RT-RPA-LFD assay showed high specificity to CTLV, and the sensitivity to CTLV was the same as that of quantitative RT-PCR at 3 × 103 copies/μL CTLV RNA transcripts. A total of 45 field tangor samples were tested using RT-RPA-LFD, RT-PCR, and RT-qPCR, and the results were consistent. The results demonstrated that the RT-RPA-LFD assay is a promising tool for rapid on-site CTLV detection.

Keywords: Citrus tatter leaf virus (CTLV); Rapid visual detection; Recombinase polymerase amplification-lateral-flow dipstick test strip.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Flexiviridae
  • Immunoassay
  • Nucleic Acid Amplification Techniques / methods
  • RNA*
  • Recombinases*
  • Sensitivity and Specificity

Substances

  • Recombinases
  • RNA

Supplementary concepts

  • Apple stem grooving virus