Rapid and sensitive detection of Singapore grouper iridovirus by loop-mediated isothermal amplification
| Autor: | Jun Gong, X.L. Mao, Qiwei Qin, Sheng Zhou, Dan Xu, Huachun Cui |
|---|---|
| Rok vydání: | 2008 |
| Předmět: |
Iridoviridae
food.ingredient DNA polymerase Iridovirus Molecular Sequence Data Loop-mediated isothermal amplification Polymerase Chain Reaction Applied Microbiology and Biotechnology law.invention food Plasmid law Virology Animals Grouper Polymerase chain reaction DNA Primers Base Sequence biology Fishes General Medicine biology.organism_classification Molecular biology DNA Viral Food Microbiology biology.protein Nucleic Acid Amplification Techniques Nested polymerase chain reaction Biotechnology |
| Zdroj: | Journal of Applied Microbiology. 105:389-397 |
| ISSN: | 1365-2672 1364-5072 |
| Popis: | Aims: The aim of this paper was to develop a loop-mediated isothermal amplification (LAMP) method for rapid, sensitive and inexpensive detection of Singapore grouper iridovirus (SGIV) in grouper (GP), Epinephelus sp. Methods and Results: A set of six specific primers was designed by targeting the SGIV ORF-014L. With Bst DNA polymerase large fragment, the target DNA can be amplified as early as 20 min at 65°C in a simple water bath. The detection limit is about 0·02 fg (equivalent to 6·3 copies) of plasmid ORF-014L. LAMP products could be judged with three different methods. There were no cross-reactions with seven other aquatic animal viruses indicating high specificity of the LAMP. The LAMP method was applied to detect SGIV in virus-infected GP cells and GP tissues effectively. Conclusions: The LAMP described in this study is a cheap, sensitive, specific and rapid protocol for the detection of SGIV in cells and in GP tissues. Significance and Impact of the Study: The developed LAMP method can be simply applied both in field condition and in laboratory operation for specific detection of SGIV infection. |
| Databáze: | OpenAIRE |
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