Abstract
Optimization of Recombinant Glycoprotein D (gD) based Indirect ELISA for Detection of Antibodies against Bovine Herpesvirus-1
Highlights
Bovine herpesvirus-1 (BoHV-1) primarily infects cattle and buffaloes and causes infectious bovine rhinotracheitis (IBR) and infectious pustular vulvovaginitis (IPV) along with other complications such as conjunctivitis, abortion, encephalitis, enteritis, and generalized disease in newborn (Muylkens et al, 2007)
The immunogenic region of glycoprotein D (gD) gene was amplified by PCR using gene specific primers, which resulted in 834bp product (Figure 1)
The expression conditions were optimized to 8h of induction with 0.15% Rhamnose at 32°C (Supplementary Figure S3).The protein was expressed in insoluble form as inclusion bodies, purified under denaturation condition by Ni-NTA purification method followed by refolding with decreasing concentration of urea and the purified expressed protein was obtained
Summary
Bovine herpesvirus-1 (BoHV-1) primarily infects cattle and buffaloes and causes infectious bovine rhinotracheitis (IBR) and infectious pustular vulvovaginitis (IPV) along with other complications such as conjunctivitis, abortion, encephalitis, enteritis, and generalized disease in newborn (Muylkens et al, 2007). The BoHV-1 is a member of the subfamily Alphaherpesvirinae which includes Human herpesvirus-1 (HHV-1), a prototype virus for Alphaherpesvirinae (Misra et al, 1981; Mayank et al, 2018). The IBR and IPV are among the endemic diseases of cattle in India as a result of crossbreeding. The virus was isolated and reported for the first time by Mehrotra et al (1976) from crossbred calves at an organized cattle herd in Uttar Pradesh, who had a history of keratoconjunctivitis. The disease has been reported in most of the states of India
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