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Probe-based real-time reverse transcription loop-mediated isothermal amplification (RRT-LAMP) assay for rapid and specific detection of foot-and-mouth disease virus
- Source :
- Transboundary and emerging diseasesREFERENCES. 67(6)
- Publication Year :
- 2019
-
Abstract
- Rapid and specific detection of foot-and-mouth disease virus (FMDV) is a key factor for promoting prompt control of FMD outbreaks. In this study, a real-time reverse transcription loop-mediated isothermal amplification (RRT-LAMP) assay with high sensitivity, rapidity and reliability was developed using a targeted gene-specific assimilating probe for real-time detection of seven FMDV serotypes. Positive assay signals were generated within 15 min for the lowest concentration of a standard RNA sample at 62°C; this was substantially faster than that achieved by the OIE (World Organisation for Animal Health)-recommended real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) assay. The new assay specifically amplified the 3D gene of all seven FMDV serotypes and did not amplify other viral nucleic acids. The detection limit of the assay was 102 copies/µl which is comparable to that achieved by qRT-PCR. Furthermore, using clinical samples, the results of the RRT-LAMP assay were largely in agreement with those from the qRT-PCR assay with a kappa value (95% confidence interval [CI]) of 0.94 (0.86-1.02). The established RRT-LAMP assay that features assimilating probes is an advanced molecular diagnostic tool that is easily applicable to a wide range of circumstances and has high potential for use as an on-site diagnostic assay for rapid, specific, and reliable detection of FMDVs in clinical samples.
- Subjects :
- 040301 veterinary sciences
Loop-mediated isothermal amplification
Real-Time Polymerase Chain Reaction
Sensitivity and Specificity
Virus
0403 veterinary science
03 medical and health sciences
Animals
Reverse Transcription Loop-mediated Isothermal Amplification
030304 developmental biology
Detection limit
0303 health sciences
General Veterinary
General Immunology and Microbiology
biology
Chemistry
Reverse Transcriptase Polymerase Chain Reaction
Reproducibility of Results
04 agricultural and veterinary sciences
General Medicine
biology.organism_classification
Virology
Reverse transcriptase
Reverse transcription polymerase chain reaction
Molecular Diagnostic Techniques
Foot-and-Mouth Disease Virus
Foot-and-Mouth Disease
Nucleic acid
Foot-and-mouth disease virus
Nucleic Acid Amplification Techniques
Subjects
Details
- ISSN :
- 18651682
- Volume :
- 67
- Issue :
- 6
- Database :
- OpenAIRE
- Journal :
- Transboundary and emerging diseasesREFERENCES
- Accession number :
- edsair.doi.dedup.....9300f8f9770ac07eaf7d56721a7b8da2