Skip to main content

Development of an enzyme-linked immunosorbent assay using recombinant protein antigen for the diagnosis of Chikungunya virus.

We describe here the development of in-house enzyme linked immunosorbent assay (ELISA) for diagnostic Chikungunya virus (CHIKV) infection using a recombinant protein of CHIKV. Recombinant protein gene was designed based on 154 sequences and we used computational methods to predict the structure and antigenic potential.


 For the prediction of confirmation, the Monkey Recombinant Proteins gene coding for a recombinant protein CHIKV (rCHIKVp) has been synthesized and expressed in a prokaryotic system. Subsequently, the protein was purified by affinity chromatography and used as an antigen in an indirect ELISA. We present data on production optimization and preparation of the recombinant antigen ELISA to detect IgG against CHIKV in human sera.
Development of an enzyme-linked immunosorbent assay using recombinant protein antigen for the diagnosis of Chikungunya virus.

Development of an enzyme-linked immunosorbent assay for viral infections in horses latex using recombinant E2 protein as an antigen.


Gum virus causes fever, skin eruptions, and limb edema in horses. For high-throughput and time-saving method for serodiagnosis, we explored Other Recombinant Proteins the immunogenic viral antigen sap, and establish an enzyme-linked immunosorbent assay (ELISA) using recombinant proteins. Western blot analysis using serum from infected horses show a strong reaction with the viral antigen of about 46 kDa in accordance with E1 or E2 glycoproteins.

 Recombinant E2 (RE2) proteins reacted more strongly with sera infected horses than re1 protein in both Western blotting and ELISA. In ELISA using protein Re2 (RE2-ELISA), for all horses experimentally infected with the virus latex (n = 7), the optical density (OD) exceeds the cutoff value at 14 days post-infection. BPO in five of the nine horses vaccinated also slightly exceeded the cutoff value after vaccination.

 Among the naturally infected horses (n = 28), 24 seronegative in acute sera, which turned out to be seropositive in sera recovered. During four seropositive horses in acute sera, serial dilution method endpoint with paired sera detect ≥4-fold increase in titer. In conclusion, we established RE2-ELISA can detect antibodies to the virus after infection experimental latex horse and nature; This should be useful in the diagnosis and surveillance of viral infections sap.
Labels:

Comments

Post a Comment

Popular posts from this blog

Optimization of recombinant Zika virus NS1 protein secretion from HEK293 cells.

Sensitive, accurate and cost-effective diagnostic tests are needed to detect the Zika virus (ZIKV) infection. Nonstructural 1 (NS1) glycoprotein is an excellent diagnostic marker because it was released in a hexameric conformation of the infected cells into the bloodstream of patients in the early course of infection.  We established a stable system Fungal Recombinant Proteins  of his rZNS1-expression in HEK293 cells through lentiviral transduction. New optimization approach to improve his rZNS1 secretion of proteins in mammalian expression systems is achieved through incubation of 50 nM rapamycin followed by incubation in serum-free media for 9 days, attaining the protein from ~ 10 mg / l of culture medium.  His rZNS1 purified hexamer recognized by anti-NS1 antibodies in serum ZIKV patients, and demonstrated the ability to induce a humoral response in mice immunized. recombinant proteins obtained are reliable biological tool that can potentially be applied in the development of diagn
Post a Comment
Read more

Recombinant Lactococcus Lactis Expressing M1-HA2 Fusion Protein Provides Protective Mucosal Immunity Against H9N2 Avian Influenza Virus in Chickens.

H9N2 subtype avian influenza virus of low pathogenicity (LPAIV) are distributed worldwide and cause enormous economic losses in the poultry industry. Although almost all the chickens immunized with the inactivated vaccine, the disease is still widespread. We speculate that increase mucosal or cellular immune response may contribute to improving the H9N2 virus surveillance.  In this study, we Bovine Recombinant Proteins  constructed a novel Lactococcus lactis (L. lactis) strain expressing a recombinant fusion protein consisting of the M1 and HA2 proteins derived from the H9N2 viral strain endemic conserved antigens. M1-HA2 fusion protein was cloned downstream of the gene encoding the secretory peptide, and we subsequently confirmed that the fusion protein is secreted from L.  lactis by Western blotting. We assess the immunogenicity and protective effects of recombinant L. lactis this tension. Eight chicken 1-day-old were divided into four groups, and the experimental group orally vacci
Post a Comment
Read more