Field evaluation of a multiplex real-time reverse transcription polymerase chain reaction assay for detection of Vesicular stomatitis virus.
Abstract: Sporadic outbreaks of vesicular stomatitis (VS) in the United States result in significant economic losses for the U.S. livestock industries because VS is a reportable disease that clinically mimics foot-and-mouth disease. Rapid and accurate differentiation of these 2 diseases is critical because their consequences and control strategies differ radically. The objective of the current study was to field validate a 1-tube multiplexed real-time reverse transcription polymerase chain reaction (real-time RT-PCR) assay for the rapid detection of Vesicular stomatitis New Jersey virus and Vesicular stomatitis Indiana virus strains occurring in Mexico and North and Central America. A comprehensive collection of 622 vesicular lesion samples obtained from cattle, horses, and swine from throughout Mexico and Central America was tested by the real-time RT-PCR assay and virus isolation. Overall, clinical sensitivity and specificity of the real-time RT-PCR were 83% and 99%, respectively. Interestingly, VS virus isolates originating from a specific region of Costa Rica were not detected by real-time RT-PCR. Sequence comparisons of these viruses with the real-time RT-PCR probe and primers showed mismatches in the probe and forward and reverse primer regions. Additional lineage-specific primers and a probe corrected the lack of detection of the missing genetic lineage. Thus, this assay reliably identified existing Mexican and Central American VS viruses and proved readily adaptable as new VS viruses were encountered. An important secondary result of this research was the collection of hundreds of new VS virus isolates that provide a foundation from which many additional studies can arise.
Publication Date: 2009-03-17 PubMed ID: 19286495DOI: 10.1177/104063870902100201Google Scholar: Lookup
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- Journal Article
- Research Support
- U.S. Gov't
- Non-P.H.S.
- Validation Study
Summary
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The research paper titled “Field evaluation of a multiplex real-time reverse transcription polymerase chain reaction assay for detection of Vesicular stomatitis virus“, discusses a method of rapidly detecting Vesicular stomatitis New Jersey virus and Vesicular stomatitis Indiana virus strains in livestock from Mexico and Central America using a multiplex real-time RT-PCR assay. Despite a few shortcomings, the assay was found to be able to adapt and identify the viruses effectively.
Overview of Research
- The researchers aimed to develop an efficient method to differentiate between foot-and-mouth disease and vesicular stomatitis, two conditions that result in significant economic loss for livestock industries in the US.
- The goal was to quickly identify these viruses, as their impact and treatment strategies differ significantly.
The Study Approach
- The researchers used a collection of 622 vesicular lesion samples from cattle, horses, and swine across Mexico and Central America to conduct their study.
- These samples were tested with both the newly-developed real-time RT-PCR assay and traditional virus isolation techniques to evaluate the real-time RT-PCR assay’s clinical sensitivity and specificity.
Study Findings
- The test showed a clinical sensitivity of 83% and a specificity of 99%.
- However, it could not detect VS viruses originating from a specific region in Costa Rica due to mismatches in the probe and primers with these virus strains.
- Upon further investigation, the researchers designed lineage-specific primers and probes to rectify the lack of detection for these variants.
Significance and Limitations
- The assay held its merit in being able to adapt to new VS viruses as and when they were encountered, suggesting potential in identifying existing VS viruses in Mexico and Central America effectively.
- An unintended, but important, byproduct of this research was the collection of several new VS virus isolates which could be used for future investigations.
- The limitation of this study lied in the initial inability of the assay to detect certain virus strains due to sequence mismatches.
- However, researchers were able to adapt the assay to rectify this.
Cite This Article
APA
Wilson WC, Letchworth GJ, Jiménez C, Herrero MV, Navarro R, Paz P, Cornish TE, Smoliga G, Pauszek SJ, Dornak C, George M, Rodriguez LL.
(2009).
Field evaluation of a multiplex real-time reverse transcription polymerase chain reaction assay for detection of Vesicular stomatitis virus.
J Vet Diagn Invest, 21(2), 179-186.
https://doi.org/10.1177/104063870902100201 Publication
Researcher Affiliations
- Plum Island Animal Disease Center, USDA, Agricultural Research Service, PO Box 848, Greenport, NY 11944, USA.
MeSH Terms
- Animals
- Animals, Domestic / virology
- Central America
- Immunohistochemistry / veterinary
- Mexico
- RNA, Viral / chemistry
- RNA, Viral / genetics
- Reverse Transcriptase Polymerase Chain Reaction / veterinary
- Sensitivity and Specificity
- Sequence Analysis, DNA
- Vesicular Stomatitis / diagnosis
- Vesicular Stomatitis / virology
- Vesicular stomatitis Indiana virus / genetics
- Vesicular stomatitis Indiana virus / isolation & purification
- Vesicular stomatitis New Jersey virus / genetics
- Vesicular stomatitis New Jersey virus / isolation & purification
Citations
This article has been cited 8 times.- Fonseca Júnior AA, Laguardia-Nascimento M, Barbosa AAS, da Silva Gonçalves VL, de Oliveira AM, Rivetti Júnior AV, Camargos MF. Phylodynamics of Alagoas vesiculovirus in Brazil. Braz J Microbiol 2022 Sep;53(3):1691-1699.
- Hole K, Nfon C, Rodriguez LL, Velazquez-Salinas L. A Multiplex Real-Time Reverse Transcription Polymerase Chain Reaction Assay With Enhanced Capacity to Detect Vesicular Stomatitis Viral Lineages of Central American Origin. Front Vet Sci 2021;8:783198.
- Drolet BS, Reeves WK, Bennett KE, Pauszek SJ, Bertram MR, Rodriguez LL. Identical Viral Genetic Sequence Found in Black Flies (Simulium bivittatum) and the Equine Index Case of the 2006 U.S. Vesicular Stomatitis Outbreak. Pathogens 2021 Jul 23;10(8).
- Brister H, Barnum SM, Reedy S, Chambers TM, Pusterla N. Validation of two multiplex real-time PCR assays based on single nucleotide polymorphisms of the HA1 gene of equine influenza A virus in order to differentiate between clade 1 and clade 2 Florida sublineage isolates. J Vet Diagn Invest 2019 Jan;31(1):137-141.
- Velazquez-Salinas L, Naik S, Pauszek SJ, Peng KW, Russell SJ, Rodriguez LL. Oncolytic Recombinant Vesicular Stomatitis Virus (VSV) Is Nonpathogenic and Nontransmissible in Pigs, a Natural Host of VSV. Hum Gene Ther Clin Dev 2017 Jun;28(2):108-115.
- Fan Q, Xie Z, Xie Z, Deng X, Xie L, Huang L, Luo S, Huang J, Zhang Y, Zeng T, Wang S, Liu J, Pang Y. Development of a GeXP-multiplex PCR assay for the simultaneous detection and differentiation of six cattle viruses. PLoS One 2017;12(2):e0171287.
- Tolardo AL, Souza WM, Romeiro MF, Vieira LC, Luna LK, Henriques DA, Araujo Jd, Siqueira CE, Colombo TE, Aquino VH, Fonseca BA, Bronzoni RV, Nogueira ML, Durigon EL, Figueiredo LT. A real-time reverse transcriptase polymerase chain reaction for detection and quantification of Vesiculovirus. Mem Inst Oswaldo Cruz 2016 Jun 3;111(6):385-90.
- Johnson N, Voller K, Phipps LP, Mansfield K, Fooks AR. Rapid molecular detection methods for arboviruses of livestock of importance to northern Europe. J Biomed Biotechnol 2012;2012:719402.
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