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 60 min. 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 1 fg of BoHV-1 DNA (dilution
of 106). 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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