The resulting standard curves showed high correlation coefficient values. The specificity of the method was validated by the absence of any cross-reaction with the RNA samples extracted from other shrimp viruses (Taura syndrome virus, white spot syndrome virus, infectious hypodermal and haematopoietic necrosis virus, hepatopancreatic parvovirus) and specific pathogen-free (SPF) shrimp. The detection sensitivity of the rRT-LAMP method was as low as 7 × 10(0) copies per reaction. The LAMP reaction was optimized to contain 8 mmol l(-1) Mg(2+) and 1♴ mmol l(-1) dNTPs, and to be performed at 58☌ for 60 min. A set of six specific primers was successfully designed targeting a conserved region of the YHV genome. In this study, a one-step, real-time reverse-transcription loop-mediated isothermal amplification (rRT-LAMP) assay was developed for better detection of both genotypes of YHV-1 and YHV-8. A new genotype of yellow head virus (YHV), designated as YHV-8, was found in farmed shrimp Fenneropenaeus chinensis suffering suspectedly from EMS/AHPNS (early mortality disease/acute hepatopancreatic necrosis disease) in China in 2012.

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