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

Abstract

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. © 2012 Elsevier B.V.