期刊
LETTERS IN APPLIED MICROBIOLOGY
卷 56, 期 6, 页码 428-435出版社
OXFORD UNIV PRESS
DOI: 10.1111/lam.12065
关键词
colorimetric; gold nanoparticle; loop-mediated isothermal amplification; Penaeus vannamei nodavirus
资金
- National Research Council of Thailand
- National Center for Genetic Engineering and Biotechnology (BIOTEC)
- Office of the Higher Education Commission (HEC)
- Mahidol University under the National Research University Initiatives
Penaeus vannamei nodavirus (PvNV) is an emerging viral infection that has caused muscle necrosis or a white tail disease in cultivated whiteleg shrimp Penaeus (Litopenaeus) vannamei, leading to loss in the productions. Rapid detection of PvNV is essential for further control disease. A combination between reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay and colorimetric gold nanoparticle (AuNP) probe was developed for rapid, sensitive and inexpensive detection of PvNV in this study. RT-LAMP reaction successfully detected PvNV at 63 degrees C for 45 and 5min for hybridization of LAMP amplified product, followed by salt-induced AuNP-labelled ssDNA probe aggregation to visual colour development. This method showed identical results to LAMP followed by gel electrophoresis and spectrophotometric detection, and it was 10 times sensitive more than conventional nested RT-PCR. The new method revealed negative results with other shrimp pathogens. This study provides the direct visualization of LAMP reaction by AuNP probe hybridization, that significantly reduces the time and cost required for the molecular diagnostic of infectious diseases and offers to use in aquaculture health management.
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