Development and evaluation of a reverse transcription loop-mediated isothermal amplification assay for rapid typing of serotype O foot-and-mouth disease virus in endemic regions of Tanzania

Abstract

AbstractFoot-and-mouth disease (FMD) caused by FMD virus, is a highly infectious viral disease affecting livestock. Diagnosis plays a vital role in disease control and management during disease outbreaks. Available serotyping approaches are costly, time-consuming, requires complex equipment and trained personnel, warranting development of a diagnostic method that addresses this gap. In this retrospective study, a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for the detection of serotype O FMDV was developed and evaluated using forty-three FMDV isolates from cattle. The specificity of the assay was tested against other FMDV isolates (serotypes A, SAT 1 and SAT 3). Only FMDV serotype O samples could be amplified within 26 min with an anneal derivative temperature of 87.02 degree celsius. Additionally, the assay amplified the viral protein 1 (VP1) gene with a detection limit of 0.0378 ng/μl. This molecular diagnostic approach has potential future application in improving FMDV surveillance as it provides baseline information for controlling FMD outbreaks in Tanzania and other Eastern and Southern African countries.