Direct detection of unamplified spring viraemia of carp virus RNA using unmodified gold nanoparticles
| Autor: | O Schachner, Mona Saleh, Hatem Soliman, Mansour El-Matbouli |
|---|---|
| Rok vydání: | 2012 |
| Předmět: |
Detection limit
Carps biology Oligonucleotide Positive reaction RNA Metal Nanoparticles Vesiculovirus Aquatic Science biology.organism_classification Rapid detection Virology Molecular biology Sensitivity and Specificity Virus Fish Diseases Viral Envelope Proteins Colloidal gold Rhabdoviridae Infections Animals RNA Viral Colorimetry Gold Carp Ecology Evolution Behavior and Systematics |
| Zdroj: | Diseases of aquatic organisms. 100(1) |
| ISSN: | 0177-5103 |
| Popis: | Spring viraemia of carp (SVC) is a viral disease that mainly affects carp Cyprinus carpio and other cyprinid fish, causing severe economic losses. Rapid detection and identification of spring viraemia of carp virus (SVCV) is crucial for effective disease management. Recent advances in nanoscience are having a significant impact on many scientific fields, especially biodiagnostics, where a number of nanoparticle-based assays have been introduced for biomolecular detection. Single- and double-stranded oligonucleotides can be adsorbed on gold nanoparticles (AuNPs) in colloidal solution under certain conditions. We exploited this phenomenon to develop a specific hybridization assay for direct detection of SVCV-RNA without prior amplification. The result of the hybridization process could be detected visually within 1 min when the colour of the reaction mixture changed from red to blue (positive reaction) or remains red (negative). The lower detection limit of the assay was estimated to be 10-3 TCID50 ml-1 SVCV-RNA, and it has the feasibility to detect the target virus-RNA in clinical specimens without previous amplification. In order to obtain an indication of the assay's performance on clinical samples we compared the optimized assay with nested RT-PCR in detection of SVCV-RNA in infected fish samples. The concordance of the 2 methods was defined as 100% when compared to nested RT-PCR positive and negative samples. The SVC-AuNPs assay requires only 15 min, eliminates the need for thermal cycling or detection instruments and is a specific and rapid tool for detection of SVCV-RNA directly from clinical samples. |
| Databáze: | OpenAIRE |
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