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Rapid and visual detection of novel astroviruses causing fatal gout in goslings using one-step reverse transcription loop-mediated isothermal amplification
- Source :
- Poultry Science, Vol 99, Iss 9, Pp 4259-4264 (2020), Poultry Science
- Publication Year :
- 2020
- Publisher :
- Elsevier, 2020.
-
Abstract
- To visually and rapidly detect a novel goose astrovirus (N-GoAstV) causing fatal gout in goslings, an isothermal detection method based on one-step reverse transcription loop-mediated isothermal amplification (one-step RT-LAMP) was established. The one-step RT-LAMP assay for N-GoAstV detection, using Bst 3.0 DNA polymerase with strong reverse transcription activity and primer sets targeting the opening reading frame 1b (ORF1b) of N-GoAstV, could be completed in 30 min using a water bath at 61°C; the detection results could be visually observed by adding a pH-sensitive dye containing phenol red and cresol red. The detection limit of the one-step RT-LAMP assay was 57.8 copies/ÎĽL, which was similar to that of reverse transcription–quantitative polymerase chain reaction. The assay specifically detected N-GoAstV without any cross-reaction with other reference viruses, and this was further confirmed using enzyme digestion. These results indicated that the newly established RT-LAMP assay could accomplish reverse transcription, amplification, and visual result determination in one step, and the results obtained via this rapid and cost-effective method could be used to support disease control on farms in terms of N-GoAstV infection.
- Subjects :
- Gout
DNA polymerase
Loop-mediated isothermal amplification
N-GoAstV
Sensitivity and Specificity
Article
law.invention
03 medical and health sciences
chemistry.chemical_compound
law
Astroviridae Infections
Geese
Animals
Reverse Transcription Loop-mediated Isothermal Amplification
Polymerase chain reaction
030304 developmental biology
lcsh:SF1-1100
Detection limit
Phenol red
0303 health sciences
biology
0402 animal and dairy science
one-step RT-LAMP
04 agricultural and veterinary sciences
General Medicine
visible detection
040201 dairy & animal science
Molecular biology
Reverse transcriptase
rapidity
chemistry
Molecular Diagnostic Techniques
biology.protein
Astroviridae
Animal Science and Zoology
lcsh:Animal culture
Primer (molecular biology)
Nucleic Acid Amplification Techniques
Subjects
Details
- Language :
- English
- ISSN :
- 00325791
- Volume :
- 99
- Issue :
- 9
- Database :
- OpenAIRE
- Journal :
- Poultry Science
- Accession number :
- edsair.doi.dedup.....318301d38df81aa71a51493bf9bff2a1