| Detection of shrimp Taura syndrome virus by loop-mediated isothermal amplification using a designed portable multi-channel turbidimeter. | |
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MedLine Citation:
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PMID: 21619895 Owner: NLM Status: Publisher |
Abstract/OtherAbstract:
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In this study, a portable turbidimetric end-point detection method was devised and tested for the detection of Taura syndrome virus (TSV) using spectroscopic measurement of a loop-mediated isothermal amplification (LAMP) by-product: magnesium pyrophosphate (Mg(2)P(2)O(7)). The device incorporated a heating block that maintained an optimal temperature of 63°C for the duration of the RT-LAMP reaction. Turbidity of the RT-LAMP by-product was measured when light from a light-emitting diode (LED) passed through the tube to reach a light dependent resistance (LDR) detector. Results revealed that turbidity measurement of the RT-LAMP reactions using this device provided the same detection sensitivity as the agarose gel electrophoresis detection of RT-LAMP and nested RT-PCR (IQ2000™) products. Cross reactions with other shrimp viruses were not found, indicating that the RT-LAMP-turbidity measurement was highly specific to TSV. The combination of 10min for rapid RNA preparation with 30min for RT-LAMP amplification followed by turbidity measurement resulted in a total assay time of less than 1h compared to 4-8h for the nested RT-PCR method. RT-LAMP plus turbidity measurement constitutes a platform for the development of more rapid and user-friendly detection of TSV in the field. |
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Authors:
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Assawapong Sappat; Wansadaj Jaroenram; Teeranart Puthawibool; Tanom Lomas; Adisorn Tuantranont; Wansika Kiatpathomchai |
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Publication Detail:
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Type: JOURNAL ARTICLE Date: 2011-5-18 |
Journal Detail:
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Title: Journal of virological methods Volume: - ISSN: 1879-0984 ISO Abbreviation: - Publication Date: 2011 May |
Date Detail:
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Created Date: 2011-5-30 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 8005839 Medline TA: J Virol Methods Country: - |
Other Details:
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Languages: ENG Pagination: - Citation Subset: - |
Copyright Information:
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Copyright © 2011. Published by Elsevier B.V. |
Affiliation:
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National Electronics and Computer Technology Center, 112 Pahol Yothin Rd., Klong Luang, Pathumthani 12120, Thailand. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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