Issue Date

2023

Abstract

Abaca bunchy top is a major disease of abaca (Musa textilis Nee) caused by babuvirus species Abaca bunchy top virus (ABTV) and Banana bunchy top virus (BBTV). These viruses have similar biological properties but differ in some molecular characteristics that would allow specific virus detection. The study was conducted to validate primers for specific detection by polymerase chain reaction (PCR), and design single-reaction specific methods for BBTV or ABTV. Validation of published primer pairs using multiple sequence alignment and in silico PCR identified four and ten primer pairs specific to BBTV and ABTV, respectively. Conventional PCR is

limited in requiring separate assays for each virus hence detection methods based on triple-primer PCR (TP- PCR) and rolling circle amplification-restriction fragment length polymorphism (RCA-RFLP) were designed

for single-reaction detection of ABTV and BBTV to circumvent this limitation. TP-PCR using the designed primers AbRF, AbRR1, and AbRR2 specifically detected BBTV and BTV in an in silico single reaction assay producing amplicons of 226 bp and 329 bp, respectively. Furthermore, the set of AgeI, AluI, BspDI, BspHI, and PstI restriction enzymes was identified and validated to yield unique restriction patterns for specific detection of BBTV and ABTV by RCA-RFLP. The specific molecular methods designed for ABTV or BBTV can be applied in efficient disease surveillance and monitoring, virus resistance screening, and indexing of abaca planting materials.

Source or Periodical Title

UP Los Baños Journal

Volume

21

Issue

2

Page

1-25

Document Type

Article

Language

English

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