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- Title
EXPRESSION OF NUCLEOCAPSID PROTEIN OF RABIES VIRUS AND ITS DIAGNOSTIC POTENTIAL.
- Authors
Rather, Mudasir M.; Upmanyu, Vikramaditya; Vasavi, Koppu; Pande, Tripti; Singh, Sudhir; Pawaskar, N. K.; Poloju, Deepa
- Abstract
Rabies virus, belonging to the Lyssavirus genus within the Rhabdoviridae family, continues to pose a significant and widespread threat to both human health and wildlife populations on a global scale. The causative agent of this disease, the rabies virus is notably neurotropic, making it particularly dangerous. Several diagnostic techniques are available, but among them, the Rapid Fluorescent Focus Inhibition Test (RFFIT) stands out as the most reliable and sensitive method. RFFIT plays a pivotal role in the laboratory diagnosis of rabies virus infection. It is an essential technique used to quantify neutralizing antibodies against the rabies virus present in a patient's blood serum, offering high specificity and sensitivity. During the execution of the RFFIT procedure, the use of anti-nucleocapsid FITC Conjugate is indispensable. This component serves as a vital element in the laboratory diagnosis of rabies virus infection. To develop anti-nucleocapsid antibodies, it is essential to undertake cloning and expression of the nucleocapsid protein which was done in the prokaryotic expression system by cloning the N-terminal fragment of the rabies virus N protein which contains major immunodominant epitopes/antigenic determinant incorporated in it. PCR amplified N gene encoding the N protein was cloned into an expression plasmid to produce the recombinant antigen in Escherichia coli cells BL21 (Rosette gami). SDS-PAGE showed the presence of the product with expected molecular weight (35 kDa). Subsequently the immunoreactivity of the expressed protein was checked by western blotting (Anti-histidine, as well as polyclonal serum). The expressed protein can then be administered in an appropriate animal model to elicit an antibody response against this recombinant protein. Subsequently, these antibodies can be conjugated with FITC dye and employed in the RFFIT process, contributing to the accurate diagnosis of rabies virus infection.
- Subjects
RABIES virus; VIRAL proteins; GENE expression; PROTEIN expression; EPITOPES; VIRUS diseases
- Publication
Journal of Experimental Zoology India, 2024, Vol 27, Issue 1, p457
- ISSN
0972-0030
- Publication type
Article
- DOI
10.51470/jez.2024.27.1.457