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Home / Equine Research Bank / Vet Rec / Real-time quantitative RT-PCR and PCR assays for a novel Eur...
The Veterinary record2007; 160(18); 611-618; doi: 10.1136/vr.160.18.611

Real-time quantitative RT-PCR and PCR assays for a novel European field isolate of equine infectious anaemia virus based on sequence determination of the gag gene.

Abstract: In 2006, an outbreak of equine infectious anaemia (EIA) occurred in Ireland. The initial source of the outbreak is believed to have been contaminated plasma imported from Italy. This paper presents the nucleotide sequence of the gag gene of the virus identified in Ireland (EIAV(Ire)), the first for a European strain of EIAV. Comparison of the gag gene with North American and Asian strains of the virus showed that the gag gene is less well conserved than previously believed, and that EIAV strains can have similar phenotypes despite considerable variations in genotype. On the basis of the deduced sequence of the EIAV(Ire) gag gene, highly sensitive, specific and quantitative RT-PCR and PCR assays were developed, and used to quantify the EIAV nucleic acid in postmortem tissues, plasma and secretions of infected horses. This is the first report of the detection and quantification of EIAV in nasal, buccal and genital swabs by RT-PCR.
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Publication Date: 2007-05-08 PubMed ID: 17483378DOI: 10.1136/vr.160.18.611Google Scholar: Lookup
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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 research paper details the creation and utilization of real-time quantitative RT-PCR and PCR assays for a new form of the equine infectious anaemia (EIA) virus, found in Ireland and identified through the sequencing of the virus’s gag gene.

Introduction

  • This study was instigated by an outbreak of EIA in Ireland in 2006, hypothesized to have originated from contaminated plasma imported from Italy.
  • The research focused on a particular gene of the EIA virus identified in Ireland, known as the gag gene. This was the first time a gag gene was sequenced in a European strain of EIA virus.

Findings

  • When compared to North American and Asian strains of the EIA virus, it was discovered that the gag gene is not as well conserved as previously thought. This implies that EIA virus strains can have similar disease manifestations despite considerable variations in their genetic makeup or genotype.
  • Based on the newly deduced sequence of the Irish EIA virus gag gene, precise and quantitative RT-PCR and PCR assays were developed.

Application of RT-PCR and PCR Assays

  • These newly crafted assays were thereafter utilized to quantify the EIA virus’s nucleic acid within postmortem tissues, plasma, and secretions from infected horses.
  • Excitingly, this study marks the first report of EIA virus detection and quantification in nasal, buccal, and genital swabs using RT-PCR. This pioneering step may provide a quicker, more targeted approach to diagnosing EIA in horses.

Conclusion

  • Ultimately, the isolation and sequencing of the EIA virus’s gag gene has led to the development of a more sensitive, specific and quantitative assay to both detect and measure the virus in horses.
  • This in-depth knowledge of the virus’s genetic makeup may improve our understanding of how this virus operates and could potentially aid in the creation of prevention methods or treatments in the future.

Cite This Article

APA
Quinlivan M, Cook RF, Cullinane A. (2007). Real-time quantitative RT-PCR and PCR assays for a novel European field isolate of equine infectious anaemia virus based on sequence determination of the gag gene. Vet Rec, 160(18), 611-618. https://doi.org/10.1136/vr.160.18.611

Publication

The Veterinary record
ISSN: 0042-4900
NlmUniqueID: 0031164
Country: England
Language: English
Volume: 160
Issue: 18
Pages: 611-618

Researcher Affiliations

Quinlivan, M
  • Virology Unit, Irish Equine Centre, Johnstown, Naas, County Kildare, Ireland.
Cook, R F
    Cullinane, A

      MeSH Terms

      • Amino Acid Sequence
      • Animals
      • Base Sequence
      • Disease Outbreaks / veterinary
      • Equine Infectious Anemia / diagnosis
      • Genotype
      • Horses
      • Infectious Anemia Virus, Equine / isolation & purification
      • Ireland / epidemiology
      • Molecular Sequence Data
      • Polymerase Chain Reaction / methods
      • Polymerase Chain Reaction / veterinary
      • Reproducibility of Results
      • Reverse Transcriptase Polymerase Chain Reaction / methods
      • Reverse Transcriptase Polymerase Chain Reaction / veterinary
      • Sensitivity and Specificity
      • Sequence Alignment / veterinary
      • Sequence Homology, Nucleic Acid
      • Species Specificity

      Citations

      This article has been cited 6 times.
      1. 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.
        doi: 10.1007/s00253-023-12504-5pubmed: 37039847google scholar: lookup
      2. Sharav T, Konnai S, Ochirkhuu N, Ts EO, Mekata H, Sakoda Y, Umemura T, Murata S, Chultemdorj T, Ohashi K. Detection and molecular characterization of equine infectious anemia virus in Mongolian horses.. J Vet Med Sci 2017 Nov 17;79(11):1884-1888.
        doi: 10.1292/jvms.17-0202pubmed: 29021424google scholar: lookup
      3. Malik P, Singha H, Goyal SK, Khurana SK, Kumar R, Virmani N, Shanmugasundaram K, Pandey SB, Kant R, Singh BK, Singh RK. Sero-surveillance of equine infectious anemia virus in equines in India during more than a decade (1999-2012).. Indian J Virol 2013 Dec;24(3):386-90.
        doi: 10.1007/s13337-013-0142-3pubmed: 24426302google scholar: lookup
      4. Singha H, Goyal SK, Malik P, Khurana SK, Singh RK. Development, evaluation, and laboratory validation of immunoassays for the diagnosis of equine infectious anemia (EIA) using recombinant protein produced from a synthetic p26 gene of EIA virus.. Indian J Virol 2013 Dec;24(3):349-56.
        doi: 10.1007/s13337-013-0149-9pubmed: 24426297google scholar: lookup
      5. Issel CJ, Scicluna MT, Cook SJ, Cook RF, Caprioli A, Ricci I, Rosone F, Craigo JK, Montelaro RC, Autorino GL. Challenges and proposed solutions for more accurate serological diagnosis of equine infectious anaemia.. Vet Rec 2013 Feb 23;172(8):210.
        doi: 10.1136/vr-2012-100735pubmed: 23161812google scholar: lookup
      6. Cappelli K, Capomaccio S, Cook FR, Felicetti M, Marenzoni ML, Coppola G, Verini-Supplizi A, Coletti M, Passamonti F. Molecular detection, epidemiology, and genetic characterization of novel European field isolates of equine infectious anemia virus.. J Clin Microbiol 2011 Jan;49(1):27-33.
        doi: 10.1128/JCM.01311-10pubmed: 21084503google scholar: lookup
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