Development of a recombinase polymerase amplification assay for the diagnosis of banana bunchy top virus in different banana cultivars

  • Reetika Kapoor
  • Nishant Srivastava
  • Shailender Kumar
  • R. K. Saritha
  • Susheel Kumar Sharma
  • Rakesh Kumar Jain
  • Virendra Kumar Baranwal
Brief Report

Abstract

Recombinase polymerase amplification (RPA) is a rapid, isothermal amplification method with high specificity and sensitivity. In this study, an assay was developed and evaluated for the detection of banana bunchy top virus (BBTV) in infected banana plants. Three oligonucleotide primer pairs were designed from the replicase initiator protein gene sequences of BBTV to function both in RPA as well as in polymerase chain reaction (PCR). A total of 133 symptomatic as well as asymptomatic banana leaf samples from various cultivars were collected from the different regions of India and evaluated for BBTV infection using the RPA assay. BBTV was efficiently detected using crude leaf sap in RPA and the results obtained were consistent with PCR-based detection using purified DNA as template. To our knowledge, this is the first report of reliable diagnosis of BBTV infection by RPA using crude leaf sap as a template.

Supplementary material

705_2017_3399_MOESM1_ESM.pdf (276 kb)
Supplementary material 1 (PDF 276 kb)

Copyright information

© Springer-Verlag Wien 2017

Authors and Affiliations

  • Reetika Kapoor
    • 1
  • Nishant Srivastava
    • 1
  • Shailender Kumar
    • 1
  • R. K. Saritha
    • 1
  • Susheel Kumar Sharma
    • 2
  • Rakesh Kumar Jain
    • 1
  • Virendra Kumar Baranwal
    • 1
  1. 1.Division of Plant Pathology, Advanced Centre for Plant VirologyIndian Agricultural Research InstituteNew DelhiIndia
  2. 2.ICAR Research Complex for NEH RegionManipur CentreImphalIndia

Personalised recommendations