Development and evaluation of an antigen-capture ELISA for detection of the UL24 antigen of the duck enteritis virus, based on a polyclonal antibody against the UL24 expression protein

Abstract

An antigen-capture enzyme-linked immunosorbent assay (AC-ELISA) method was developed for the efficient detection of the UL24 antigen of the duck enteritis virus (DEV) using polyclonal antibodies. Ducks and rabbits were immunized, respectively, with expressed UL24 recombinant protein. The IgG antibodies against UL24 from ducks and rabbits were purified and used as the capture antibodies. The specificity of the optimized AC-ELISA was evaluated by use of DEV, duck hepatitis virus (DHV), duck hepatitis B virus (DHBV), gosling plague virus (GPV), Riemerella anatipestifer (R.A.), Escherichia coli (E. coli), Pasteurella multocida (P.M.) and Salmonella Enteritidis (S.E.). Only DEV specimens yielded a specific and strong signal. The limit of the sensitivity of this method for the detection of DEV was 46 ng/100 microl. Compared with PCR and virus isolation, the rate of agreement for the detection of experimentally infected sera was 100%. A comparative test used on clinical specimens between the neutralization test and the AC-ELISA showed that the proportions of true positives and true negatives by the AC-ELISA were 0.90 and 0.67 respectively. These results indicated that the AC-ELISA approach is rapid, sensitive, and reliable for specific detection of DEV antigen.

Two nuclear localization signals regulate intracellular localization of the Duck enteritis virus UL13 protein

“…The experimental operation was carried out as described previously [70][71][72]. Briefly, the full-length UL13 sequence was PCR amplified from DEV chromosomal DNA using the primers F-UL13F and F-UL13R.…”

Two nuclear localization signals regulate intracellular localization of the Duck enteritis virus UL13 protein

“…Putative nuclear localization signals (NLSs) within the UL13 coding sequence were identified using PSORT II Prediction (http://psort.nibb.ac.jp/form2.html) [74]. Sequence analysis using PSORT II predicted that UL13 has two potential NLSs in its arginine-rich regions, namely, RRRR at aa 4 to 7 and PGKRKTK at aa 90 to 96 ( Figure S1).…”

The Role of VP16 in the Life Cycle of Alphaherpesviruses

“…For all herpesviruses, a complete virion consists of four parts: a core that contains a double-stranded DNA genome, a capsid, a tegument, and an envelope (Guiping et al, 2007;Boštíková et al, 2014). The alphaherpesvirinae subfamily includes herpes simplex virus type-1/2 (HSV-1/2), pseudorabies virus (PRV), duck enteritis virus (DEV) (Qi et al, 2008;Zhao et al, 2008;Chang et al, 2009;Guo et al, 2009;Jia et al, 2009), varicella-zoster virus (VZV), equine herpesviruses (EHV), bovine herpesvirus (BHV), canine herpesvirus (CHV), and Marek's disease virus (MDV) (Kobty, 2015;Smith, 2017;You et al, 2017;He et al, 2018;Lefkowitz et al, 2018).…”