Back to Search
Start Over
Rapid and sensitive detection of infectious spleen and kidney necrosis virus by loop-mediated isothermal amplification combined with a lateral flow dipstick.
- Source :
-
Archives of virology [Arch Virol] 2010 Mar; Vol. 155 (3), pp. 385-9. Date of Electronic Publication: 2010 Jan 27. - Publication Year :
- 2010
-
Abstract
- Loop-mediated isothermal amplification (LAMP) allows rapid amplification of nucleic acids under isothermal conditions. In this report, a 20-min LAMP amplification of the DPOL gene of infectious spleen and kidney necrosis virus (ISKNV) using a biotin-labeled primer was combined with lateral flow dipstick (LFD) chromatography for rapid and simple visual detection of ISKNV-specific amplicons. The LFD process involves a 5-min specific hybridization with an FITC-labeled DNA probe to confirm the presence of complement ISKNV amplicons that were biotinated in LAMP. The resulting DNA duplexes, consisting of labeled probes and amplicons, migrate along the LFD strip by chromatography for 5 min and are trapped at the test line and visualized by biotin labeling. The detection limit of ISKNV by LAMP-LFD was 10 copies. The results show that the LAMP-LFD method has the advantages of better sensitivity and speed and less dependence on equipment than the standard PCR for specifically detecting low levels of ISKNV DNA, and this can be useful in the field as a routine diagnostic tool.
- Subjects :
- Animals
DNA Primers genetics
DNA Virus Infections diagnosis
DNA Virus Infections virology
DNA, Viral genetics
Fish Diseases virology
Iridoviridae genetics
Molecular Sequence Data
Sensitivity and Specificity
Sequence Analysis, DNA
Time Factors
DNA Virus Infections veterinary
Fish Diseases diagnosis
Iridoviridae isolation & purification
Molecular Diagnostic Techniques methods
Nucleic Acid Amplification Techniques methods
Subjects
Details
- Language :
- English
- ISSN :
- 1432-8798
- Volume :
- 155
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- Archives of virology
- Publication Type :
- Academic Journal
- Accession number :
- 20107846
- Full Text :
- https://doi.org/10.1007/s00705-010-0593-4