Rapid and simple colorimetric loop-mediated isothermal amplification (LAMP) assay for the detection of Bovine alphaherpesvirus 1

As the causative agent of Infectious Bovine Rhinotracheitis (IBR) and Pustular Vulvovaginitis/Balanoposthitis (IPV/IPB), alphaherpesvirus 1 (BoHV-1) is responsible for high economic losses in cattle industry worldwide. This study aimed to establish a fast, colorimetric loop-mediated isothermal amplification (LAMP) assay detection viral DNA. Phenol red used as pH-sensitive readout, relying on distinct color change from pink yellow case positive reaction. LAMP reactions with different primers were compared newly designed set targeting gene encoding tegument protein V67 provided best results, enabling readout within 8?30 min. showed less cross-reactions other ruminant alphaherpesviruses than qPCR but was 10-fold sensitive. However, still detected down 14 copies. The test performance evaluated using 26 well-characterized nasal swabs respiratory disease. All samples correctly identified when column-extracted Using simple DNA precipitation method, only two weak-positive turned indeterminate. Combining this makeshift water bath heated by gastronomic immersion heater allowed successful application under resource-limited conditions. technique can therefore help managing IBR/IPV outbreaks where sophisticated laboratory equipment unavailable.