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Title: Detection of shrimp Taura syndrome virus by loop-mediated isothermal amplification using a designed portable multi-channel turbidimeter
Authors: Assawapong Sappat
Wansadaj Jaroenram
Teeranart Puthawibool
Tanom Lomas
Adisorn Tuantranont
Wansika Kiatpathomchai
Thailand National Electronics and Computer Technology Center
Mahidol University
Thailand National Center for Genetic Engineering and Biotechnology
Keywords: Immunology and Microbiology
Issue Date: 1-Aug-2011
Citation: Journal of Virological Methods. Vol.175, No.2 (2011), 141-148
Abstract: 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. © 2011 Elsevier B.V.
ISSN: 18790984
Appears in Collections:Scopus 2011-2015

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