Peer-reviewed veterinary case report
Rapid detection of BoHV-1 genomic DNA by loop-mediated isothermal amplification assay.
- Journal:
- Journal of virological methods
- Year:
- 2014
- Authors:
- El-Kholy, Alaa A et al.
- Affiliation:
- Veterinary Serum & Vaccine Research Institute
Plain-English summary
Bovine herpes virus-1 (BoHV-1) is a serious virus that affects both domestic and wild cattle. Researchers have created a new test called the BoHV-1-LAMP assay, which can quickly and accurately detect this virus in samples. This test works by using a special enzyme and a set of six designed primers to amplify the virus's DNA at a specific temperature for an hour. It has been shown to be very sensitive, able to detect even tiny amounts of the virus, and can distinguish between different types of BoHV-1. Overall, this new test is a promising tool for better managing and controlling BoHV-1 infections in cattle.
Abstract
Bovine herpes virus-1 (BoHV-1) is a serious viral pathogen of domestic and wild cattle. Herein, we report development of a new molecular diagnostic assay for rapid and sensitive detection of BoHV-1 utilizing the loop-mediated isothermal amplification (LAMP) technique. BoHV-1-LAMP assay was optimized to amplify the target DNA by incubation the Bst-DNA polymerase enzyme with a set of specially constructed six primers, based on the gE-gene of BoHV-1 virus, at 65°C for 60min. BoHV-1-LAMP products were detected by visual inspection using SYBR Green-I stain and had a ladder-like appearance by gel electrophoresis analysis. Negative results obtained with DNA from other tested fish viruses confirmed the specificity of the assay. The analytical sensitivity of the BoHV-1-LAMP assay was 1fg of BoHV-1 DNA (dilution of 10(6)). The developed assay could successfully detect BoVH-1 DNA from clinical samples. Results of this study indicate that the developed BoHV-1-LAMP is rapid and highly sensitive assay not only for detection of BoHV-1 in clinical samples, but also for differentiation between wild-type (gE-positive) and gE-negative BoHV-1 viruses, which will improve the control programs of BoHV-1 in Egypt.
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Search related cases →Original publication: https://pubmed.ncbi.nlm.nih.gov/24769199/