Recombinant Salmonella enterica OmpX protein expression and its potential for serologically diagnosing Salmonella abortion in mares.
Abstract: abortion in mares is caused by subspecies serovar infection and is characterized by premature (abortion) or non-viable fetus birth. Although all horses are susceptible to infection, the condition is more often clinically manifested in pregnant mares, with most abortions recorded in young females. In addition, nonspecific clinical disease signs and poorly sensitive and effective bacteriological diagnostic methods hinder rapid and reliable infection diagnoses. Immunochemical methods such as enzyme-linked immunosorbent assay (ELISA) and immunochromatography assays can facilitate effective and rapid diagnoses. However, they require highly specific and active antigens and antibodies. This study aimed to generate a recombinant outer membrane protein X (OmpX) and evaluate its suitability for serological diagnosis of abortion in mares Unassigned: Outer membrane protein X from the antigen was synthesized and expressed in using the pET28 vector. Transformed cells were cultured under different conditions to detect recombinant OmpX (rOmpX) expression, and rOmpX purification and refolding were both conducted using metal affinity chromatography. Refolded and purified rOmpX was characterized by western blotting, liquid chromatography with tandem mass spectrometry, and ELISA. Unassigned: After optimized rOmpX expression, a 23 kDa molecular weight protein was identified. Amino acid sequence analysis using Mascot program suggested that these peptides were the OmpX protein from . High specificity and diagnostic efficiency were recorded when rOmpX was used in ELISA against 89 serum samples from aborted and contact mares. Unassigned: Recombinant outer membrane protein, in comparison to the O antigen, demonstrated better diagnostic characteristics against sera from mares who aborted and contact horses.
Copyright: © Borovikov, et al.
Publication Date: 2023-09-13 PubMed ID: 37859952PubMed Central: PMC10583879DOI: 10.14202/vetworld.2023.1790-1795Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
This study explores a more effective and quick method to diagnose a disease caused by the bacteria Salmonella enterica that provokes abortions in mares. The researchers generated a protein model from the bacteria and tested it via serological methods proving it to be a promising diagnostic tool.
Creation of Recombinant Protein
- Researchers began by creating a synthetic version of the outer membrane protein X, commonly referred to as OmpX, found in the Salmonella enterica bacterium.
- To express this protein, they inserted the synthetic gene into the E. coli bacteria using the pET28 vector, a device commonly used in bioengineering to produce proteins.
- The bacteria were then grown under various conditions to see how well OmpX was expressed.
- The expressed OmpX was then extracted and purified using a process called metal affinity chromatography which helps separate and collect individual protein molecules.
Evaluation of Recombinant Protein
- The researchers validated the success of the extraction and purification process by using western blotting, an analytical technique that helps detect specific proteins, and liquid chromatography with tandem mass spectrometry, which identifies specific compounds in a sample.
- They also utilized ELISA, a commonly used antibody test, to confirm the presence of OmpX.
Testing the Recombinant Protein for Diagnostic Efficiency
- After successfully expressing and purifying the recombinant OmpX, the researchers used this protein in an ELISA test on 89 serum samples taken from aborted and contact mares to ascertain its diagnostic efficiency.
- Analysis of the results showed that the new protein model has high specificity and diagnostic efficiency and performed better than the pre-existing antigen.
Conclusion
- The rOmpX protein presented promising diagnostic capacities and could serve as a new, effective tool for early detection and diagnosis of Salmonella abortion in mares.
- This not only allows for prompt medical intervention but also helps in limiting the spread of the disease.
Cite This Article
APA
Borovikov S, Ryskeldina A, Tursunov K, Syzdykova A, Akibekov O.
(2023).
Recombinant Salmonella enterica OmpX protein expression and its potential for serologically diagnosing Salmonella abortion in mares.
Vet World, 16(9), 1790-1795.
https://doi.org/10.14202/vetworld.2023.1790-1795 Publication
Researcher Affiliations
- Department of Microbiology and Biotechnology, Faculty of Veterinary and Animal Husbandry Technology, S. Seifullin Kazakh Agrotechnical Research University, 010000, Astana, Kazakhstan.
- Department of Veterinary Medicine, Faculty of Veterinary and Animal Husbandry Technology, S. Seifullin Kazakh Agrotechnical Research University, 010000, Astana, Kazakhstan.
- Laboratory of Immunochemistry and Immunobiotechnology, National Center for Biotechnology, 010000, Astana, Kazakhstan.
- Research Platform of Agricultural Biotechnology, S. Seifullin Kazakh Agrotechnical Research University, 010011, Astana, Kazakhstan.
- Department of Microbiology and Biotechnology, Faculty of Veterinary and Animal Husbandry Technology, S. Seifullin Kazakh Agrotechnical Research University, 010000, Astana, Kazakhstan.
Conflict of Interest Statement
The authors declare that they have no competing interests.
References
This article includes 21 references
- Uzzau S, Brown D.J, Wallis T, Rubino S, Leori G, Bernard S, Casadesús J, Platt D.J, Olsen J.E. Host adapted serotypes of Salmonella enterica.. Epidemiol. Infect. 2000;125(2):229–255.
- Grandolfo E, Parisi A, Ricci A, Lorusso E, de Siena R, Trotta A, Buonavoglia D, Martella V, Corrente M. High mortality in foals associated with Salmonella enterica subsp. enterica Abortusequi infection in Italy.. J. Vet. Diagn. Invest. 2018;30(3):483–485.
- . OIE Terrestrial Manual-Salmonellosis. Chapter 3.10.7.. 2022:1–7.
- Wang J, Guo K, Li S, Liu D, Chu X, Wang Y, Guo W, Du C, Wang X, Hu Z. Development and application of real-time PCR assay for detection of Salmonella abortusequi.. J. Clin. Microbiol. 2023;61(3):e0137522.
- Ren Y, Wei J, Wang Y, Wang P, Ji Y, Liu B, Wang J, González-Sapienza G, Wang Y. Development of a streptavidin-bridged enhanced sandwich ELISA based on self-paired nanobodies for monitoring multiplex Salmonella serogroups.. Anal. Chim. Acta. 2022;1203:339705.
- Burgess B.A, Noyes N.R, Bolte D.S, Hyatt D.R, van Metre D.C, Morley P.S. Rapid Salmonella detection in experimentally inoculated equine faecal and veterinary hospital environmental samples using commercially available lateral flow immunoassays.. Equine Vet. J. 2015;47(1):119–122.
- Sotnikov D.V, Barshevskaya L.V, Zherdev A.V, Eskendirova S.Z, Mukanov K.K, Mukantayev K.K, Ramankulov Y.M, Dzantiev B.B. Immunochromatographic system for serodiagnostics of cattle brucellosis using gold nanoparticles and signal amplification with quantum dots.. Appl. Sci. 2020;10(3):738.
- Minicozzi J, Sanchez S, Lee M.D, Holt P.S, Hofacre C.L, Maurer J.J. Development of recombinant flagellar antigens for serological detection of Salmonella enterica Serotypes Enteritidis, Hadar, Heidelberg andTyphimurium in poultry.. Agriculture. 2013;3(3):381–397.
- Makvandi M, Teimoori A, Parsa N.M, Khodadadi A, Cheshmeh M.G.D, Zandi M. Expression of Salmonella Typhimurium and Escherichia coli flagellin protein and its functional characterization as an adjuvant.. Microb. Pathog. 2018;118:87–90.
- Dehghani B, Rasooli I, Gargari S.L.M, Nadooshanb M.R.J, Owliac P, Nazarian S. Immunogenicity of Salmonella enterica serovar Enteritidis virulence protein, InvH, and cross-reactivity of its antisera with Salmonella strains.. Microbiol. Res. 2013;168(2):84–90.
- Li M, Fang H, Fan X, Zhang J, Yan J, Sun A. Distribution of Salmonella paratyphi A outer membrane protein X gene and immune-protective effect related to its recombinant expressed products.. Chin. J. Epidemiol. 2013;34(12):1219–1222.
- Ferrer-Navarro M, Ballesté-Delpierre C, Vila J, Fàbrega A. Characterization of the outer membrane subproteome of the virulent strain Salmonella Typhimurium SL1344.. J. Proteomics. 2016;146:141–147.
- Laemmli U.K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.. Nature. 1970;227(5259):680–685.
- Borovikov S, Tursunov K, Syzdykova A, Begenova A, Zhakhina A. Expression of recombinant Omp18 and MOMP of Campylobacter jejuni and the determination of their suitability as antigens for serological diagnosis of campylobacteriosis in animals.. Vet. World. 2023;16(1):222–228.
- Müller-Loennies S, Brade L, Brade H. Neutralizing and cross-reactive antibodies against enterobacterial lipopolysaccharide.. Int. J. Med. Microbiol. 2007;297(5):321–340.
- Hajam I.A, Dar P.A, Sekar S.C, Nanda R, Kishore S, Bhanuprakash V, Ganesh K. Expression, purification, and functional characterisation of flagellin, a TLR5-ligand.. Vet. Ital. 2013;49(2):181–186.
- Mirhosseini S.A, Fooladi A.A.I, Amani J, Sedighian H. Production of recombinant flagellin to develop ELISA-based detection of Salmonella enteritidis.. Braz. J. Microbiol. 2017;48(4):774–781.
- Fitzgerald G.A, Komarov A, Kaznadzey A, Mazo I, Kireeva M.L. Expression of SARS-CoV-2 surface glycoprotein fragment 319–640 in E. coli, and its refolding and purification.. Protein Expr. Purif. 2021;183:105861.
- Zhansaya A, Malika N, Boris D, Kanat T, Kanatbek M, Yerlan R, Kasym M. Expression of recombinant CTLA-4 and PD-L1 proteins fused with thioredoxin, and determination of their ligand-binding activities.. Rep. Biochem. Mol. Biol. 2022;11(2):310–319.
- Meyer T, Stratmann-Selke J, Meens J, Schirrmann T, Gerlach G.F, Frank R, Dübel S, Strutzberg-Minder K, Hust M. Isolation of scFv fragments specific to OmpD of Salmonella Typhimurium.. Vet. Microbiol. 2011;147(1–2):162–169.
- Li Q, Zhu Y, Yin K, Xu L, Yin C, Li Y, Ren J, Yuan Y, Jiao X. Purification of recombinant IpaJ to develop an indirect ELISA-based method for detecting Salmonella enterica serovar Pullorum infections in chickens.. BMC Vet. Res. 2019;15(1):3.
Citations
This article has been cited 0 times.Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
