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Peer-reviewed veterinary case report

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

Journal:
Virus research
Year:
2014
Authors:
Liu, Xuemei et al.
Affiliation:
College of Veterinary Medicine · China

Plain-English summary

Researchers in China have noticed an increase in cases of a virus called hemorrhagic enteritis virus (HEV) affecting yellow chickens. They developed a new test called the RealAmp assay that can quickly diagnose this virus in just 15 minutes using a portable device. This test is very sensitive and only detects HEV, making it reliable for both field and lab use. When compared to another testing method called nested PCR, the results matched perfectly. Overall, the RealAmp assay is a promising tool for identifying this virus in chickens.

Abstract

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 15min, using a simple and portable device, the ESE-Quant Tube Scanner. The detection limit of DNA was 1fg/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.

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Original publication: https://pubmed.ncbi.nlm.nih.gov/24487182/