Detection of African horsesickness virus in infected spleens by a sandwich ELISA using two monoclonal antibodies specific for VP7.
Abstract: A sandwich enzyme-linked immunsorbent assay (ELISA) for rapid detection of African horsesickness virus (AHSV) in infected spleens or cell culture supernatant was developed. This method uses two monoclonal antibodies (MAbs) which recognize two non-overlapping epitopes of the major core protein (VP7) to coat the solid phase, and one labeled with biotin as second antibody. This ELISA was evaluated for its ability to detect AHSV in infected spleens resulting in a sensitivity of 97.4% and a specificity of 100% compared with virus isolation in cell culture, and can be used for the detection of the nine different AHSV serotypes.
Publication Date: 1992-08-01 PubMed ID: 1517353DOI: 10.1016/0166-0934(92)90113-rGoogle Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
- African Horse Sickness
- Antibodies
- Cell Culture
- Diagnosis
- Diagnostic Technique
- Disease
- Disease Diagnosis
- Enzyme-Linked Immunosorbent Assay (ELISA)
- Epidemiology
- Equine Diseases
- Equine Health
- Horses
- Immune System
- Infection
- Infectious Disease
- Laboratory Methods
- Monoclonal Antibodies
- Serotypes
- Veterinary Medicine
- Virology
- Virus
Summary
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The article presents a method for quickly detecting African horsesickness virus (AHSV) in infected spleens or cell cultures using a sandwich enzyme-linked immunsorbent assay (ELISA) technique. The test uses two monoclonal antibodies that interact with two distinct regions of a specific protein, showing high sensitivity and specificity.
Research Overview
- The research focuses on the development of a sandwich enzyme-linked immunsorbent assay (ELISA) test for detecting African horsesickness virus (AHSV) in infected spleens or cell culture supernatants. ELISA tests are commonly used in medical laboratories owing to their sensitivity and specificity in detecting various pathogens.
- The unique aspect of this test is that it uses two monoclonal antibodies (MAbs) that recognize two distinct, non-overlapping epitopes, or antigenic sites, of a major core protein called VP7 in the virus. These antibodies are designed to bind to these specific areas, allowing for a more accurate detection of the virus.
Methodology and Results
- The ELISA procedure involves coating the solid phase—or the surface of the test—with two types of MAbs against the AHSV specific protein (VP7). One of these antibodies is also labeled with biotin as a secondary antibody, which aids in detection.
- Through this approach, the researchers were able to detect AHSV in infected spleens with a sensitivity rate of 97.4%, meaning that it correctly identified positive cases this percentage of the time.
- In terms of specificity, the test scored a perfect 100%, meaning it correctly identified all negative cases without any false positives. Such a high specificity rate is crucial in diagnostics as it ensures that the chances of misdiagnosis are extremely low.
- Moreover, the established ELISA was proven to be efficient for differentiating among the nine different serotypes of AHSV, further enhancing its potential application in diagnostics.
Conclusion and Applications
- This research contributes to the field of veterinary diagnostics by presenting a rapid, accurate, and reliable methodology for detecting AHSV in infected spleens or cell cultures.
- Beyond its academic implications, the research holds practical significance as it can assist in early detection and prevention of the spread of African horsesickness, a highly fatal viral disease in horses.
Cite This Article
APA
Laviada MD, Babín M, Dominguez J, Sánchez-Vizcaíno JM.
(1992).
Detection of African horsesickness virus in infected spleens by a sandwich ELISA using two monoclonal antibodies specific for VP7.
J Virol Methods, 38(2), 229-242.
https://doi.org/10.1016/0166-0934(92)90113-r Publication
Researcher Affiliations
- Instituto Nacional de Investigaciones Agrarias, Departamento de Sanidad Animal, Madrid, Spain.
MeSH Terms
- African Horse Sickness / diagnosis
- African Horse Sickness Virus / isolation & purification
- Animals
- Antibodies, Monoclonal
- Antibodies, Viral
- Antigens, Viral
- Avidin
- Biotin
- Capsid / immunology
- Capsid / isolation & purification
- Capsid Proteins
- Enzyme-Linked Immunosorbent Assay / methods
- Horses
- Sensitivity and Specificity
- Spleen / microbiology
Citations
This article has been cited 8 times.- Fowler VL, Howson ELA, Flannery J, Romito M, Lubisi A, Agüero M, Mertens P, Batten CA, Warren HR, Castillo-Olivares J. Development of a Novel Reverse Transcription Loop-Mediated Isothermal Amplification Assay for the Rapid Detection of African Horse Sickness Virus. Transbound Emerg Dis 2017 Oct;64(5):1579-1588.
- Bachanek-Bankowska K, Maan S, Castillo-Olivares J, Manning NM, Maan NS, Potgieter AC, Di Nardo A, Sutton G, Batten C, Mertens PP. Real time RT-PCR assays for detection and typing of African horse sickness virus. PLoS One 2014;9(4):e93758.
- van Wyngaardt W, Mashau C, Wright I, Fehrsen J. Serotype- and serogroup-specific detection of African horsesickness virus using phage displayed chicken scFvs for indirect double antibody sandwich ELISAs. J Vet Sci 2013;14(1):95-8.
- Aradaib IE, Mohemmed ME, Sarr JA, Idris SH, Ali NO, Majid AA, Karrar AE. A simple and rapid method for detection of African horse sickness virus serogroup in cell cultures using RT-PCR. Vet Res Commun 2006 Apr;30(3):319-24.
- Martínez-Torrecuadrada JL, Díaz-Laviada M, Roy P, Sánchez C, Vela C, Sánchez-Vizcaíno JM, Casal JI. Serologic markers in early stages of African horse sickness virus infection. J Clin Microbiol 1997 Feb;35(2):531-5.
- Stone-Marschat M, Carville A, Skowronek A, Laegreid WW. Detection of African horse sickness virus by reverse transcription-PCR. J Clin Microbiol 1994 Mar;32(3):697-700.
- Saliki JT, House JA, Mebus CA, Dubovi EJ. Comparison of monoclonal antibody-based sandwich enzyme-linked immunosorbent assay and virus isolation for detection of peste des petits ruminants virus in goat tissues and secretions. J Clin Microbiol 1994 May;32(5):1349-53.
- Sakamoto K, Punyahotra R, Mizukoshi N, Ueda S, Imagawa H, Sugiura T, Kamada M, Fukusho A. Rapid detection of African horsesickness virus by the reverse transcriptase polymerase chain reaction (RT-PCR) using the amplimer for segment 3 (VP3 gene). Arch Virol 1994;136(1-2):87-97.
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