Genetic targets for the detection and identification of Venezuelan equine encephalitis viruses.
Abstract: Rt-PCR probes targeted to different gene sequences of VEE (Venezuelan equine encephalitis) virus strain TC-83 were assessed for their sensitivity, specificity and non-specific cross-reactivity. A generic VEE virus amplimer (VNSP4F2/VNSP4R2), targeted against nsP4 was identified, which was sensitive (detected at least 10 pfu) and robust (worked over a wide range of salt concentrations and annealing temperatures). An E2 amplimer designed against TC-83, (VE2F/VE2R), identified VEE strains TRD (1AB), P676 (1C), 3880 (1D) Everglades (2) vRNA whilst a second E2 primer pair designed against strain 68U201, (68UF/68UR), identified all the remaining VEE viruses in the sero-complex. This would suggest that the VEE virus E2 gene can be sub-divided at the genetic level into two separate groups making it a useful target for differentiation of serosubtypes 1 and 2 from the other VEE virus subtypes. Using a panel of amplimers targeted to different VEE genes and strains it was possible to distinguish between most of the serotypes, but most importantly, we were able to detect the epizootic strains TRD and P676 as well as other VEE viruses implicated in human disease (sero-subtypes 1D and 1E).
Publication Date: 1998-06-25 PubMed ID: 9638144DOI: 10.1007/s007050050326Google Scholar: Lookup
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- Journal Article
Summary
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This research work focuses on employing Rt-PCR probes targeted at various genetic sequences of the Venezuelan equine encephalitis (VEE) virus to detect and identify different strains. It highlighted an effective method for distinguishing between various VEE strains and implicates the potential to detect those involved in human illnesses.
Research Details
- Time-reverse transcription polymerase chain reaction (Rt-PCR) probes focused on different genetic sequences from the Venezuelan equine encephalitis (VEE) virus, a strain known as TC-83, were evaluated with the aim of reviewing their sensitivity, specificity, and level of non-specific cross-reactivity.
- A particular VEE virus amplimer known as VNSP4F2/VNSP4R2, targeted against nsP4, was identified as being both sensitive (it was able to detect at least 10 plaque-forming units (pfu)) and robust (it functioned efficiently despite varying salt concentrations and annealing temperatures).
- An E2 amplimer that was designed against TC-83 effectively identified a variety of VEE strains. Conversely, there was also another E2 primer pair designed against a different strain (68U201) that identified all remaining VEE viruses in the sero-complex.
Findings and Implications
- This research suggested that the VEE virus E2 gene can be genetically split into two separate groups, which makes it a valuable target for distinguishing between serosubtypes 1 and 2 from other VEE virus subtypes.
- By using a panel of amplimers targeted at different VEE genes and strains, differentiation between most of the serotypes was possible. More importantly, the researchers could detect the epizootic strains known as TRD and P676. These strains, along with others (sero-subtypes 1D and 1E), have been associated with human disease, thereby underscoring the importance of this research.
- The study’s findings could lead to enhanced strategies for the diagnosis and management of this virus, which represent potential hazards to human health.
Cite This Article
APA
Brightwell G, Brown JM, Coates DM.
(1998).
Genetic targets for the detection and identification of Venezuelan equine encephalitis viruses.
Arch Virol, 143(4), 731-742.
https://doi.org/10.1007/s007050050326 Publication
Researcher Affiliations
- CBD Porton Down, Salisbury, Wiltshire, U.K.
MeSH Terms
- Animals
- Chlorocebus aethiops
- DNA Probes
- Encephalitis Virus, Venezuelan Equine / genetics
- Encephalitis Virus, Venezuelan Equine / isolation & purification
- Evaluation Studies as Topic
- Genes, Viral
- Humans
- Polymerase Chain Reaction / methods
- Sensitivity and Specificity
- Vero Cells
Citations
This article has been cited 3 times.- de Morais Bronzoni RV, Baleotti FG, Ribeiro Nogueira RM, Nunes M, Moraes Figueiredo LT. Duplex reverse transcription-PCR followed by nested PCR assays for detection and identification of Brazilian alphaviruses and flaviviruses.. J Clin Microbiol 2005 Feb;43(2):696-702.
- Linssen B, Kinney RM, Aguilar P, Russell KL, Watts DM, Kaaden OR, Pfeffer M. Development of reverse transcription-PCR assays specific for detection of equine encephalitis viruses.. J Clin Microbiol 2000 Apr;38(4):1527-35.
- Meissner JD, Huang CY, Pfeffer M, Kinney RM. Sequencing of prototype viruses in the Venezuelan equine encephalitis antigenic complex.. Virus Res 1999 Oct;64(1):43-59.
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