Rapid Detection of Infectious Hypodermal and Hematopoietic Necrosis Virus（IHHNV） by Real-time Fluorescence Loop-mediated Isothermal Amplification Assay

Key words: 
• Infectious hypodermal and hematopoietic necrosis virus /
• real-time loop-mediated isothermal amplification assay /
• ESE-Quant tube scanner

Abstract: Infectious hypodermal and hematopoietic necrosis virus（ IHHNV） causes large economic losses to the shrimp aquaculture industry in the world. It is necessary to develop a fast,easy and accurate method to detect IHHNV. In this study,we developed a real-time fluorescence loop-mediated isothermal amplification assay for IHHNV detection on a small ESE-Quant Tube Scanner device and evaluated its sensitivity,specificity and rapidity. This method had a detection limit of 106 dilution template of genomic DNA within 30 min under isothermal condition at 63 C,and 24 copies of plasmid were detected. Furthermore,the approach had no non-specific signal response to other DNAs,including those of 27 species of aquatic animals and penaeid shrimp pathogens. This method had a positive detection rate of 59. 87%,similar to that of the real-time PCR but higher than other detection methods,and hence has great application potential in on-the-spot detection of IHHNV in shrimps.