Publication: Rapid and sensitive detection of shrimp infectious myonecrosis virus using a reverse transcription loop-mediated isothermal amplification and visual colorogenic nanogold hybridization probe assay
10
Issued Date
2013-11-01
Resource Type
ISSN
18790984
01660934
01660934
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2-s2.0-84882797843
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Mahidol University
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SCOPUS
Bibliographic Citation
Journal of Virological Methods. Vol.193, No.2 (2013), 542-547
Suggested Citation
Narong Arunrut, Jantana Kampeera, Rungkarn Suebsing, Wansika Kiatpathomchai Rapid and sensitive detection of shrimp infectious myonecrosis virus using a reverse transcription loop-mediated isothermal amplification and visual colorogenic nanogold hybridization probe assay. Journal of Virological Methods. Vol.193, No.2 (2013), 542-547. doi:10.1016/j.jviromet.2013.07.017 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/31842
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Title
Rapid and sensitive detection of shrimp infectious myonecrosis virus using a reverse transcription loop-mediated isothermal amplification and visual colorogenic nanogold hybridization probe assay
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Abstract
This study reports a novel strategy for the detection of reverse transcription loop-mediated isothermal amplification (RT-LAMP) products derived from infectious myonecrosis virus (IMNV), causes a serious myonecrosis in Penaeus (. Litopenaeus) vannamei, by using a ssDNA-labeled with gold nanoparticle (AuNP) probe. This technique relies on a self-aggregation method, when the AuNP aggregation is induced by an increasing of salt concentrations with visual detection. The presence of IMNV-LAMP target prevented an AuNP aggregation and a solution remained as pink color of AuNP, while non-complementary targets cannot prevent AuNP aggregation, resulting in a visible color change to purple color after addition of salt. By using the combination of LAMP and AuNP probe system, the total assay interval required approximately 50. min (exclude RNA preparation). Detection limit was 10. copies of IMNV RNA in vitro transcript that comparable to that of LAMP followed by LFD and nested RT-PCR, but it was 100-times more sensitive than RT-PCR methods. This assay can be adapted easily for rapid detection of other shrimp infectious diseases agents at low-cost with robust reagents and using a simple colorimetric detection method. © 2013 Elsevier B.V.