Document Type : Original Articles
Authors
1
Department of Microbiology, Falavarjan Branch, Islamic Azad University, Isfahan, Iran
2
Department of Microbiology, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran
3
Department of Microbiology, falavarjan Branch, Islamic Azad University, Isfahan, Iran
4
Department of Human vaccine , Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran
5
Department of Immunology, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran
10.32592/ARI.2024.79.5.1099
Abstract
Leptospirosis is a disease that threatens the health of humans and animals, which is caused by pathogenic Leptospira. Early detection of pathogenic Leptospires prevents many problems. LipL41-OmpL1, a protected outer membrane protein (OMP) of pathogenic Leptospira, was inserted into E. coli bacteria using different software for the amino acid sequence of OmpL1 and LipL41 to design a recombinant fusion protein and then expressed to investigate immunogenicity. The selected genes were propagated and cloned as a fusion in pET32a+ plasmid vector and expressed by Escherichia coli plays S (DE3) by heat shock method. It was evaluated in the BALB/c mouse laboratory animal model. Recombinant LipL41-OmpL1 protein was confirmed using urea purification method and western blot and its immunogenicity was evaluated by measuring high humoral immune stimulation and antibody secretion in BALB/c mice by ELISA method. The results showed that animals that received both OmpL1 and LipL41 proteins had 85% immunogenicity, while the control group that did not receive the fusion protein had only 25% immunogenicity (P < 0.001). Furthermore, no evidence of infection was found in recipients of the OmpL1-LipL41 fusion protein, suggesting that this protein is safe to use. The protective effects of immunization with OmpL1 and LipL41 were synergistic, as significant levels of protection were not observed in animals immunized with OmpL1 or LipL41 alone. Overall, this study highlights the potential of recombinant OmpL1 and LipL41 fusion protein as a promising research strategy on the development of vaccines and ELISA diagnostic kit for prevention and rapid diagnosis of leptospirosis.
Keywords
Main Subjects