Development of a bead-based assay for detection of three banana-infecting viruses

Cheng-Ping Kuan; Chia-Hsin Tsai; Ching-Shan Tseng; Tso-Chi Yang

doi:10.7717/peerj.13409

Development of a bead-based assay for detection of three banana-infecting viruses

PeerJ. 2022 May 26:10:e13409. doi: 10.7717/peerj.13409. eCollection 2022.

Authors

Cheng-Ping Kuan¹, Chia-Hsin Tsai², Ching-Shan Tseng¹, Tso-Chi Yang¹

Affiliations

¹ Division of Biotechnology, Taiwan Agricultural Research Institute, Taichung, Taiwan.
² Division of Plant Pathology, Taiwan Agricultural Research Institute, Taichung, Taiwan.

Abstract

Background: Banana bunchy top virus (BBTV), cucumber mosaic virus (CMV) and banana streak virus (BSV) are important banana viruses, there are possible infections frequently with several viruses in field. Since the viruses are readily trasmitted in vegetative propagules, which pose a threat to banana production in banana-growing areas.

Methods: A multiplex polymerase chain reaction (PCR) protocol combined with LiquiChip analysis to identify BSV, BBTV, and CMV, with consistent amplification of plant ubiquitin (UBQ), the banana plant messenger RNA used as a procedural control. Multiplex reverse transcription (RT)-PCR amplicons were extended by allele-specific primers, followed by hybridization with carboxylated microspheres containing unique fluorescent oligonucleotides, which were detected using the LiquiChip 200 workstation.

Results: In this study, we aimed to develop a rapid, sensitive, and simultaneous detection method for BSV, BBTV, and CMV using a bead-based multiplex assay that can be applied in routine diagnosis. We demonstrated that this detection system was extremely efficient and highly specialized for differentiating individual in a mixture of viruses while being ten times more sensitive than traditional RT-PCR. The development of this method makes it feasible to detect banana viruses in field collected leaf samples.

Keywords: Banana; Banana bunchy top virus; Banana streak virus; Cucumber mosaic virus; Detection.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Babuvirus*
Cucumovirus* / genetics
Cytomegalovirus Infections*
Multiplex Polymerase Chain Reaction
Musa*
Plant Diseases

Supplementary concepts

Banana streak virus

Grants and funding

This research was supported by grants from the Council of Agriculture (108AS-7.6.2-CI-C1). There was no additional external funding received for this study. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.