Rapid and sensitive detection of shrimp yellow head virus using loop-mediated isothermal amplification and a colorogenic nanogold hybridization probe

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• 作者：Wansadaj Jaroenram^a ; ^b ; Narong Arunrut^a ; ^c ; Wansika Kiatpathomchai^a ; ^c ; ^{wansika@biotec.or.th}
• 关键词：YHV ; RT-LAMP ; Gold nanoparticle ; Colorogenic nanogold hybridization
• 刊名：Journal of Virological Methods
• 出版年：2012
• 出版时间：December, 2012
• 年：2012
• 卷：186
• 期：1-2
• 页码：36-42
• 全文大小：1475 K

文摘

Salt-induced self-aggregation of gold nanoparticles (AuNP) carrying unisense ssDNA probes can be prevented specifically by complementary DNA. Loop-mediated isothermal amplification (LAMP) can amplify DNA rapidly. Here, the two techniques were combined to detect yellow head virus (YHV). The LAMP-AuNP method required 60 min for LAMP and 5 min for hybridization of LAMP products to an AuNP-labeled ssDNA probe followed by salt induced probe-particle aggregation to visualize color development. The detection sensitivity of the method was comparable to that of the commercial IQ2000? nested RT-PCR but only required ¡«65 min to produce a result, and did not cross-detect other shrimp viruses. As the LAMP-AuNP protocol only requires a heating block, it offers opportunities for rapid detection of YHV.