Development of recombinant capsid antigen/transmembrane epitope fusion proteins for serological diagnosis of animal lentivirus infections.
Abstract: Among animal lentiviruses, Feline immunodeficiency virus (FIV), Equine infectious anaemia virus (EIAV) and Small ruminant lentiviruses (SRLV) are important pathogens associated with a variety of clinical pictures including immunodeficiency, anaemia, arthritis, pneumonia. The detection of viral antibody response represents a practical diagnostic approach in all lentivirus infections since they remain detectable long life. Capsid antigen (CA) is the major viral core protein and specific antibodies against this antigen are usually first recognised in infected sheep, goat and horse, remaining detectable for long period. Transmembrane (TM) domain of envelope glycoprotein contains a well conserved motif known to form an immunodominant epitope in several lentiviruses. In this study a simple strategy was developed to express the entire CA and the TM epitope in a single fusion protein from equine, feline and small ruminant lentiviruses in prokaryotic system and evaluated the diagnostic utility of a purified preparation in an indirect ELISA for each of the three infections. Results demonstrate that, for FIV and SRLV infections, the combination of CA and TM fractions increases the sensitivity of diagnostic tests based only on CA. The corresponding CA/TM antigen from EIAV showed excellent agreement with Coggins test.
Publication Date: 2004-09-08 PubMed ID: 15350735DOI: 10.1016/j.jviromet.2004.06.001Google Scholar: Lookup
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- Evaluation Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The study focuses on developing recombinant antigen fusion proteins for diagnosing animal lentivirus infections, including Feline immunodeficiency virus (FIV), Equine infectious anaemia virus (EIAV), and Small ruminant lentiviruses (SRLV). The researchers used a simple strategy to express the entire capsid antigen and the transmembrane epitope in a single fusion protein and then evaluated the diagnostic utility of this purified preparation in an indirect ELISA for each of the three infections.
Objective of the Study
- The research paper’s goal was to design and evaluate the diagnostic utility of a single fusion protein made up of an entire capsid antigen and the transmembrane epitope for three major animal lentivirus infections.
Significance of Determining Antibodies
- The presence of viral antibodies in animals’ blood serves as a crucial diagnostic method for all lentivirus infections.
- Antibodies against the capsid antigen (CA), the primary viral core protein, are the first to be identified in infected animals and can be detected over a long period.
Creation of Fusion Proteins
- The researchers developed a straightforward technique to express both the entire CA and the transmembrane (TM) epitope in a single fusion protein derived from the lentiviruses that affect horses, cats, and small ruminants.
- This fusion protein was then purified and tested in an indirect ELISA (Enzyme-linked immunosorbent assay).
Finding Results
- The result of this study showed that the combination of CA and TM fractions increases the sensitivity of diagnostic tests based only on CA for FIV and SRLV infections.
- The corresponding CA/TM antigen from EIAV showed excellent agreement with the Coggins test, a specific test used to detect the presence of EIAV.
Conclusion
- This approach provides a practical and effective means of diagnosing important lentivirus infections in animals and may lead to more rapid and accurate diagnostic procedures.
Cite This Article
APA
Rosati S, Profiti M, Lorenzetti R, Bandecchi P, Mannelli A, Ortoffi M, Tolari F, Ciabatti IM.
(2004).
Development of recombinant capsid antigen/transmembrane epitope fusion proteins for serological diagnosis of animal lentivirus infections.
J Virol Methods, 121(1), 73-78.
https://doi.org/10.1016/j.jviromet.2004.06.001 Publication
Researcher Affiliations
- Dipartimento di Produzioni Animali, Epidemiologia ed Ecologia, Facoltà di Medicina Veterinaria, Università di Torino, Via Leonardo da Vinci 44, 10095 Grugliasco (TO), Italy. sergio.rosati@unito.it
MeSH Terms
- Amino Acid Motifs
- Animals
- Antibodies, Viral / blood
- Antigens, Viral / genetics
- Antigens, Viral / immunology
- Capsid Proteins / genetics
- Capsid Proteins / immunology
- Cats
- Enzyme-Linked Immunosorbent Assay
- Epitopes / genetics
- Epitopes / immunology
- Equine Infectious Anemia / diagnosis
- Feline Acquired Immunodeficiency Syndrome / diagnosis
- Goat Diseases / diagnosis
- Goats
- Horses
- Immunodeficiency Virus, Feline / isolation & purification
- Infectious Anemia Virus, Equine / isolation & purification
- Lentivirus / immunology
- Lentivirus / isolation & purification
- Lentivirus Infections / diagnosis
- Lentivirus Infections / veterinary
- Lentivirus Infections / virology
- Lentiviruses, Ovine-Caprine / isolation & purification
- Protein Structure, Tertiary
- Recombinant Fusion Proteins / immunology
- Viral Envelope Proteins / chemistry
- Viral Envelope Proteins / genetics
- Viral Envelope Proteins / immunology
Citations
This article has been cited 9 times.- Hu Z, Guo K, Du C, Sun J, Naletoski I, Chu X, Lin Y, Wang X, Barrandeguy M, Samuel M, Wang W, Lau PI, Wernery U, Raghavan R, Wang X. Development and evaluation of a blocking ELISA for serological diagnosis of equine infectious anemia.. Appl Microbiol Biotechnol 2023 May;107(10):3305-3317.
- He J, Zhu S, Zhou J, Jiang W, Yin L, Su L, Zhang X, Chen Q, Li X. Rapid detection of SARS-CoV-2: The gradual boom of lateral flow immunoassay.. Front Bioeng Biotechnol 2022;10:1090281.
- Michiels R, Roels S, Vereecke N, Mathijs E, Mostin L, De Regge N. Species-Specific Humoral Immune Responses in Sheep and Goats upon Small Ruminant Lentivirus Infections Inversely Correlate with Protection against Virus Replication and Pathological Lesions.. Int J Mol Sci 2021 Sep 11;22(18).
- Cavalera S, Colitti B, Rosati S, Ferrara G, Bertolotti L, Nogarol C, Guiotto C, Cagnazzo C, Denina M, Fagioli F, Di Nardo F, Chiarello M, Baggiani C, Anfossi L. A multi-target lateral flow immunoassay enabling the specific and sensitive detection of total antibodies to SARS COV-2.. Talanta 2021 Feb 1;223(Pt 1):121737.
- Molaee V, Otarod V, Abdollahi D, Lühken G. Lentivirus Susceptibility in Iranian and German Sheep Assessed by Determination of TMEM154 E35K.. Animals (Basel) 2019 Sep 15;9(9).
- Teixeira BM, Taniwaki SA, Menezes PMM, Rodrigues AKPP, Mouta AN, Arcebispo TLM, Braz GF, da Cruz JCM, Brandão PE, Heinemann MB, Silva MX, Hosie MJ. Feline immunodeficiency virus in Northern Ceará, Brazil.. JFMS Open Rep 2019 Jul-Dec;5(2):2055116919859112.
- Li Y, Zhou F, Li X, Wang J, Zhao X, Huang J. Development of TaqMan-based qPCR method for detection of caprine arthritis-encephalitis virus (CAEV) infection.. Arch Virol 2013 Oct;158(10):2135-41.
- Grego E, Lacerenza D, Arias RR, Profiti M, Rosati S. Serological characterization of the new genotype E of small ruminant lentivirus in Roccaverano goat flocks.. Vet Res Commun 2009 Sep;33 Suppl 1:137-40.
- Brinkhof J, van Maanen C. Evaluation of five enzyme-linked immunosorbent assays and an agar gel immunodiffusion test for detection of antibodies to small ruminant lentiviruses.. Clin Vaccine Immunol 2007 Sep;14(9):1210-4.
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