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An improved reverse transcription loop-mediated isothermal amplification assay for sensitive and specific detection of serotype O foot-and-mouth disease virus
- Source :
- Journal of virological methods. 260
- Publication Year :
- 2018
-
Abstract
- A sensitive and specific swarm primer-based reverse transcription loop-mediated isothermal amplification (sRT-LAMP) assay for the detection of serotype O foot-and-mouth disease virus (FMDV) was developed and evaluated. The assay specifically amplified the VP3 gene of serotype O FMDV, but did not amplify the VP3 gene of other serotype FMDVs or any other viruses. The limit of detection of the assay was 102 TCID50/mL or 103 RNA copies/μL, which is 100 times lower than that of the RT-LAMP assay without swarm primers. The new assay is 10 times more sensitive than reverse transcription-polymerase chain reaction (RT-PCR) and is comparable to the sensitivity of real time RT-PCR (qRT-PCR). Evaluation of the assay using different serotypes of FMDV strains showed 100% agreement with the RT-PCR results. The previously reported serotype O FMDV-specific RT-LAMP assay did not detect 20 out of 22 strains of serotype O FMDVs, probably due to multiple mismatches between the primer and template sequences, showing that it is not suitable for detecting the serotype O FMDVs circulating in Pool 1 region countries, including Korea. In contrast, the developed sRT-LAMP assay with improved primers can rapidly and accurately diagnose serotype O FMDVs circulating in Pool 1 region countries and will be a useful alternative to RT-PCR and qRT-PCR.
- Subjects :
- 0301 basic medicine
Serotype
040301 veterinary sciences
Base Pair Mismatch
viruses
Loop-mediated isothermal amplification
Biology
Serogroup
Sensitivity and Specificity
Virus
0403 veterinary science
03 medical and health sciences
Limit of Detection
Virology
Republic of Korea
Animals
Reverse Transcription Loop-mediated Isothermal Amplification
Gene
DNA Primers
Reverse Transcriptase Polymerase Chain Reaction
04 agricultural and veterinary sciences
Reverse Transcription
biology.organism_classification
Reverse transcriptase
030104 developmental biology
Foot-and-Mouth Disease Virus
Foot-and-Mouth Disease
Capsid Proteins
Primer (molecular biology)
Foot-and-mouth disease virus
Nucleic Acid Amplification Techniques
Subjects
Details
- ISSN :
- 18790984
- Volume :
- 260
- Database :
- OpenAIRE
- Journal :
- Journal of virological methods
- Accession number :
- edsair.doi.dedup.....94c744045fe304a52abf412038de8ce4