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BioMed research international2016; 2016; 8543204; doi: 10.1155/2016/8543204

A Quantitative Real-Time RT-PCR Assay for the Detection of Venezuelan equine encephalitis virus Utilizing a Universal Alphavirus Control RNA.

Abstract: (VEEV) is an from the family that causes epizootic outbreaks in equids and humans in Central and South America. So far, most studies use conventional reverse transcriptase PCR assays for the detection of the different VEEV subtypes. Here we describe the development of a TaqMan quantitative real-time reverse transcriptase PCR assay for the specific detection and quantitation of all VEEV subtypes which uses in parallel a universal equine encephalitis virus control RNA carrying target sequences of the three equine encephalitis viruses. The control RNA was used to generate standard curves for the calculation of copy numbers of viral genome of (EEEV), (WEEV), and VEEV. The new assay provides a reliable high-throughput method for the detection and quantitation of VEEV RNA in clinical and field samples and allows a rapid differentiation from potentially cocirculating EEEV and WEEV strains. The capability to detect all known VEEV variants was experimentally demonstrated and makes this assay suitable especially for the surveillance of VEEV.
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Publication Date: 2016-11-29 PubMed ID: 28042576PubMed Central: PMC5153510DOI: 10.1155/2016/8543204Google 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 presents the development of a new method called TaqMan quantitative real-time reverse transcriptase PCR assay for detecting and quantifying all subtypes of Venezuelan equine encephalitis virus (VEEV) that can affect equids and humans in Central and South America. This method also instantly differentiates VEEV from other similar viruses.

Development of the TaqMan Quantitative Real-Time RT-PCR Assay

  • The paper offers a new technique designed specifically to detect and quantify all subtypes of Venezuelan equine encephalitis virus (VEEV). This virus has been largely studied using conventional reverse transcriptase PCR assays.
  • The new technique, called TaqMan quantitative real-time reverse transcriptase PCR assay, brings in an innovative approach utilizing a universal equine encephalitis virus control RNA. This RNA carries target sequences of three equine encephalitis viruses: Eastern equine encephalitis virus (EEEV), Western equine encephalitis virus (WEEV), and VEEV.
  • The control RNA was deployed to create standard graphs or curves for calculating the copy numbers of viral genome of EEEV, WEEV, and VEEV.

Advantages of the New Assay

  • It enables a fast and reliable high-throughput method for the detection and quantitation of VEEV RNA, making it a significant game-changer in diagnosing VEEV infections.
  • The assay allows for quick differentiation from potentially concurrent strains of EEEV and WEEV.
  • The ability to detect all the known VEEV variants was perseveringly validated, proving that this assay may be particularly suited for the surveillance of VEEV.
  • The assay can be conveniently applied to clinical and field samples, facilitating quick and efficient diagnosis and tracking of VEEV.

Significance of the Study

  • The development and implementation of this assay could significantly improve how VEEV infections are diagnosed and monitored.
  • This breakthrough has strong potential to enhance our understanding of VEEV distribution and variation, leading to better prevention strategies and treatment options.

Cite This Article

APA
Vina-Rodriguez A, Eiden M, Keller M, Hinrichs W, Groschup MH. (2016). A Quantitative Real-Time RT-PCR Assay for the Detection of Venezuelan equine encephalitis virus Utilizing a Universal Alphavirus Control RNA. Biomed Res Int, 2016, 8543204. https://doi.org/10.1155/2016/8543204

Publication

BioMed research international
ISSN: 2314-6141
NlmUniqueID: 101600173
Country: United States
Language: English
Volume: 2016
Pages: 8543204
PII: 8543204

Researcher Affiliations

Vina-Rodriguez, Ariel
  • Institute for Novel and Emerging Infectious Diseases, Friedrich-Loeffler-Institut, Greifswald, Insel Riems, Germany.
Eiden, Martin
  • Institute for Novel and Emerging Infectious Diseases, Friedrich-Loeffler-Institut, Greifswald, Insel Riems, Germany.
Keller, Markus
  • Institute for Novel and Emerging Infectious Diseases, Friedrich-Loeffler-Institut, Greifswald, Insel Riems, Germany.
Hinrichs, Winfried
  • Department of Molecular Structural Biology, Institute for Biochemistry, University of Greifswald, Greifswald, Germany.
Groschup, Martin H
  • Institute for Novel and Emerging Infectious Diseases, Friedrich-Loeffler-Institut, Greifswald, Insel Riems, Germany.

MeSH Terms

  • Alphavirus / genetics
  • Animals
  • Encephalitis Virus, Eastern Equine / genetics
  • Encephalitis Virus, Eastern Equine / isolation & purification
  • Encephalitis Virus, Venezuelan Equine / genetics
  • Encephalitis Virus, Venezuelan Equine / isolation & purification
  • Encephalitis Virus, Western Equine / genetics
  • Encephalitis Virus, Western Equine / isolation & purification
  • Encephalomyelitis, Venezuelan Equine / diagnosis
  • Encephalomyelitis, Venezuelan Equine / genetics
  • Encephalomyelitis, Venezuelan Equine / virology
  • Horses / virology
  • Humans
  • RNA, Viral / genetics
  • RNA, Viral / isolation & purification
  • Real-Time Polymerase Chain Reaction
  • South America

Conflict of Interest Statement

The authors declare that they have no competing interests.

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Citations

This article has been cited 7 times.
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