In the present study, a panel of polymerase chain reactions (PCRs) was compared to conventional diagnostic assays [virus isolation( VI) and indirect immunofluorescence (IFA)] used for detection of bovine herpesvirus-1 (BoHV-1) in nasal swabs collected from calves vaccinated with BoHV-1 gE negative live vaccine (n=18) and from clinically suspected calves (n=159) at different localities of Egypt (El-Sharquia, El-Salhia, El-Qalubeia and El-Behera). PCR assays were carried out utilizing specific primer sets, each targeting one of the viral genes encoding for glycoproteins D (gD), gE and gB (first round and nested gB-based PCR) as well as the thymidine kinase (tk) gene of BoHV-1. Specimens from both control and live BoHV-1 gE negative vaccinated calves were negative for all assays. Whereas, specimens from clinically suspected calves gave variable results as follows: 6/159 (3.7%), 5/159 (3.1%), 15/159 (9.4%), 15/159 (9.4%), 15/159 (9.4%), 18/159 (11.3%), and 16/159 (10.0%) were BoHV-1 positive using VI, IFA, gD-based PCR, tk-based PCR, gE-based PCR, gB- first round PCR and gB- nested PCR, respectively. In conclusion, PCR assays were more sensitive and independent of sample quality than VI or IFA. Moreover, the gE- based PCR was proved reliable for distinction between BoHV-1 gE negative-vaccinated and infected animals. Obtained data emphasized the utility of the BoHV-1 gB nested-PCR as a sensitive, discriminative and rapid diagnostic tool for epidemiological studies and control programs of BoHV-1 infections. Accordingly, it is recommended for routine screening of not only local but also imported animals and biologics for BoHV-1 contamination.