A TaqMan-based RT-qPCR assay for serotyping of Southern African territories (SAT) 2 strains of Foot-and-Mouth disease virus (FMDV) in Southern Africa

ObjectiveDetermining the serotype of circulating virus strains is important in implementing effective vaccination. In this study, Foot-and-Mouth Disease (FMD) Southern African territory 2 (SAT2) specific primers and TaqMan probe were designed towards rapid SAT2 detection and serotyping. The primers...

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Bibliographic Details
Published in:BMC research notes 2023-11, Vol.16 (1), p.1-323, Article 323
Main Authors: Kabelo, Tshephang I., Fana, Elliot M., Kesamang, Monamodi, Hyera, Joseph, Lebani, Kebaneilwe
Format: Article
Language:English
Subjects:
Design
Efficiency
Enzyme-linked immunosorbent assay
FMDV
Foot & mouth disease
Genomes
Immunization
Laboratories
Polymerase chain reaction
Primers
Research Note
Reverse transcription
RT-qPCR
SAT2
Serotypes
Serotyping
Strains (organisms)
Temperature
Thermal cycling
Vaccines
VP1 protein
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Summary:ObjectiveDetermining the serotype of circulating virus strains is important in implementing effective vaccination. In this study, Foot-and-Mouth Disease (FMD) Southern African territory 2 (SAT2) specific primers and TaqMan probe were designed towards rapid SAT2 detection and serotyping. The primers were tested by endpoint reverse transcription (RT) polymerase chain reaction (PCR) and quantitative PCR (RT-qPCR) using the vaccine strain SAT2035. The SAT2 serotype-specific RT-qPCR assay was compared with currently used ELISA and VP1 sequencing using Cohen’s kappa statistics.ResultsThe primers yielded amplicons of band size 190 bp during endpoint RT-PCR. When coupled with the probe, the primers reaction efficiency was determined to be 99% with an r2 value of 0.994. The results show that the SAT2 assay has comparable performance to VP1 sequencing (k = 1) and a moderate degree of agreement with ELISA (k = 0.571). The data shows that the newly designed assay could be considered for serotyping of SAT2 strains. However, for this assay to be complete there is a need to design effective SAT1 and SAT3 primers and probes that can be multiplexed to target other serotypes that co-circulate within relevant FMD endemic pools. For future implementation of the assay there is also a need to increase the number of field samples towards validation of the assay.
ISSN:1756-0500
1756-0500
DOI:10.1186/s13104-023-06586-7