Summary
Porcine Deltacoronavirus (PDCoV)
is an emergent swine coronavirus that infects epithelia cells from the
small intestine, and inducing watery diarrhea, vomiting and dehydration.
Clinical signs are more aggressive in piglets causing high mortality
rates (>40%) representing serious economic losses. Despite
the importance of PDCoV as an emerging coronavirus, little is known
about the currently prevalence in México. We select from GenBank a group
of 138 sequences and obtained a consensus PDCoV membrane protein
(M-PDCoV) sequence of 216 a.a. A Maximum Likelihood phylogenetic tree
was constructed and evaluate the relationship between the 138 sequences.
Also, a protein tertiary structure analysis was performed to analyze and
compare the topological differences. The phylogeny and the tertiary
structure analysis showed that M-PDCoV is highly conserved and therefore
suitable to use as an antigen in a diagnostic system. Hence, an
expression system performed in E. coli BL21 (DE3) using the
vector pET-SUMO with a His-tag was prepared, resulting in a synthetic M
gene of 654 pb to produce a recombinant M-PDCoV protein
(r M-PDCoV). Western blot test
confirmed
the r M-PDCoV immune
detection in 8 of 17 sera samples. The r M-PDCoV was able to
successfully stimulate immune response in immunized mice to produce
antibodies after day 7 of immunization (P < 0.001). Our
results show that r M-PDCoV is suitable to use in diagnostic
systems like an ELISA.