Development and evaluation of a one-step loop-mediated isothermal amplification for detection of spring viraemia of carp virus
| Autor: | Yong Teng, Congxin Xie, Y. Jiang, H. Liu, Zongxiao Liu, L. Gao, F. Tian, J. Q. Lv, Z. Chu, Haotian Li, Xiayang Xie |
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| Rok vydání: | 2008 |
| Předmět: |
Detection limit
Genes Viral biology Serial dilution Reverse Transcriptase Polymerase Chain Reaction Loop-mediated isothermal amplification Aquaculture Vesiculovirus General Medicine biology.organism_classification Sensitivity and Specificity Applied Microbiology and Biotechnology Virology Reverse transcriptase Virus Fish Diseases Rhabdoviridae Infections Specific primers Animals Viremia Aquaculture industry Carp Nucleic Acid Amplification Techniques Biotechnology |
| Zdroj: | Journal of Applied Microbiology. 105:1220-1226 |
| ISSN: | 1365-2672 1364-5072 |
| DOI: | 10.1111/j.1365-2672.2008.03858.x |
| Popis: | Aim: Spring viraemia of carp virus (SVCV) is the causative agent of SVC disease. The main aim of our study was to develop a one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for rapid, sensitive and effective detection of SVCV. Methods and Results: A set of four specific primers, two outer and two inner primers were designed based on the SVCV M gene for RT-LAMP assay. The sensitivity and specificity of RT-LAMP were determined and clinical test was performed under optimized amplification conditions (64°C, 60 min). The results showed that the assay has a high specificity and the detection limit was 80 copies using 10-fold series dilutions of SVCV RNA, 10 times more sensitive than nest reverse transcription-polymerase chain reaction. In the detection of 472 fish samples, this assay showed excellent agreement with the standard virus isolation method (κ = 0·807). Conclusions: A sensitive and specific RT-LAMP assay was successfully developed to monitor and detect SVCV. Significance and Impact of the Study: This work provides a robust method for evaluating the risk of SVCV. Given the advantages of LAMP in the detection of SVCV, this method can be applied to diagnose other viruses, which pose serious threats to the aquaculture industry. |
| Databáze: | OpenAIRE |
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