Real-time fluorescence loop-mediated isothermal amplification for the diagnosis of hemorrhagic enteritis virus

•A one-step, economical, RealAmp assay for the rapid diagnosis of HEV was developed.•RealAmp assay and nested PCR assay were performed on the field and experimental samples.•The coincidence rate of the two methods was 100%.

Suspected cases of hemorrhagic enteritis associated with hemorrhagic enteritis virus (HEV) are becoming more frequent among yellow chickens in the Guangdong Province of China. In this study, we have developed a one-step, ecumenical, real-time fluorescence loop-mediated isothermal amplification (RealAmp) assay for the rapid diagnosis of HEV. The RealAmp assay was performed at 63 °C and reduced the assay time to 15 min, using a simple and portable device, the ESE-Quant Tube Scanner. The detection limit of DNA was 1 fg/μl, and the detection was specific only to HEV. We also used nested PCR to evaluate the application of the RealAmp assay. The coincidence rate of the two methods was 100%. Our data indicated that the RealAmp assay provides a sensitive, specific, and user-friendly diagnostic tool for the identification and quantification of HEV for field diagnosis and in laboratory research.