Rapid detection of duck hepatitis virus type-1 by reverse transcription loop-mediated isothermal amplification

A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for the detection of duck hepatitis virus type-1 (DHV-1) was established. Using primers specific to the highly conserved 3D gene of DHV-1, the developed RT-LAMP assay detected the viral RNA of DHV-1 extracted from both allantoic fluid and liver samples of infected ducks. The assay is as sensitive as RT-PCR, and shows no cross-reaction with other common avian viral and bacterial pathogens. In addition to detection via ethidium bromide staining following gel electrophoresis, naked-eye observation after staining with SYBR Green I dye can be used to detect RT-LAMP products; this enables field application of this assay. The findings demonstrate that RT-LAMP can serve as a helpful tool for the detection and surveillance of DHV-1 in the poultry industry.

► RT-LAMP for duck hepatitis virus serotype 1 was established and could detect virus from allantoic fluid and liver samples.
► RT-LAMP with 1.45 pg of viral RNA resulted in clear, specific amplification.
► RT-LAMP products could be detected by naked-eye observation which enables field application of this assay.
► An incubation of 40 min yielded clear, specific products.