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Combination of an unbiased amplification method and a resequencing microarray for detecting and genotyping equine arteritis virus.
Abstract: This study shows that an unbiased amplification method applied to equine arteritis virus RNA significantly improves the sensitivity of the real-time reverse transcription-quantitative PCR (RT-qPCR) recommended by the World Organization for Animal Health. Twelve viral RNAs amplified using this method were hybridized on a high-density resequencing microarray for effective viral characterization.
Copyright © 2015, American Society for Microbiology. All Rights Reserved.
Publication Date: 2014-10-22 PubMed ID: 25339390PubMed Central: PMC4290937DOI: 10.1128/JCM.01935-14Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
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 study highlights the efficacy of using an unbiased amplification method on equine arteritis virus RNA in heightening the sensitivity of the RT-qPCR. By hybridizing the amplified viral RNAs on a high-density resequencing microarray, accurate viral characterization can be achieved.
Objective and Methodology
- The objective of this study was to explore the impact of an unbiased amplification method on RNA of the equine arteritis virus in terms of enhancing the sensitivity of the RT-qPCR, a method advised by the World Organization for Animal Health for virus detection.
- The research depended on the use of an unbiased amplification method for twelve viral RNAs and the subsequent hybridization of these amplified RNAs on a high-density resequencing microarray.
Unbiased Amplification Method
- The unbiased amplification method provides a way to enhance the detection of an entire range of nucleic acids from a specific sample, thus, maximizing the chance of capturing the viral genetic material, in this case, equine arteritis virus RNA.
Real-Time Reverse Transcription-Quantitative PCR (RT-qPCR)
- This is a diagnostic technique that helps in detecting and quantifying RNA targets. The World Organization for Animal Health, among others, endorses the use of RT-qPCR for detection of various infectious agents, including the equine arteritis virus.
- Raising the sensitivity of RT-qPCR makes for a more efficient diagnostic tool, helping to ensure that even low levels of viral RNA are effectively detected, thus enhancing the reliability of the test.
Introduction of High-density Resequencing Microarray
- A high-density resequencing microarray allows simultaneous detection of multiple genetic markers. It provides an extensive overview of the viral genetic material, thus aiding in detailed characterization of viruses.
- The use of this technology in the study, where amplified viral RNAs were hybridized on the microarray, enabled the effective characterization of the equine arteritis virus.
Implication of Research
- This study underlines the potential improvements to the sensitivity of the RT-qPCR when using an unbiased amplification method and resequencing microarray. This improved method could potentially guide detection and genotyping of various viruses, helping to manage viral infections in equine species more effectively.
Cite This Article
APA
Hans A, Gaudaire D, Manuguerra JC, Leon A, Gessain A, Laugier C, Berthet N, Zientara S.
(2014).
Combination of an unbiased amplification method and a resequencing microarray for detecting and genotyping equine arteritis virus.
J Clin Microbiol, 53(1), 287-291.
https://doi.org/10.1128/JCM.01935-14 Publication
Researcher Affiliations
- ANSES-Dozulé Laboratory for Equine Diseases, Virology Unit, Goustranville, France aymeric.hans@anses.fr.
- ANSES-Dozulé Laboratory for Equine Diseases, Virology Unit, Goustranville, France.
- Institut Pasteur, Unité Environnement et Risques Infectieux, Cellule d'Intervention Biologique d'Urgence (CIBU), Paris, France.
- Frank Duncombe Laboratory, Animal Health Department, IFR146 ICORE, Caen, France.
- Institut Pasteur, Unité d'Epidémiologie et Physiopathologie des Virus Oncogènes, Paris, France Centre National de la Recherche Scientifique, UMR 3569, Paris, France.
- ANSES-Dozulé Laboratory for Equine Diseases, Virology Unit, Goustranville, France.
- Institut Pasteur, Unité d'Epidémiologie et Physiopathologie des Virus Oncogènes, Paris, France Centre National de la Recherche Scientifique, UMR 3569, Paris, France.
- Université Paris-Est, Anses Maisons-Alfort Laboratory for Animal Health, UMR1161 Virologie, Maisons-Alfort, France.
MeSH Terms
- Animals
- Arterivirus Infections / diagnosis
- Arterivirus Infections / veterinary
- Arterivirus Infections / virology
- Equartevirus / classification
- Equartevirus / genetics
- Genotyping Techniques / methods
- Horse Diseases / diagnosis
- Horse Diseases / virology
- Horses
- Oligonucleotide Array Sequence Analysis / methods
- Phylogeny
- Virology / methods
References
This article includes 14 references
- Cavanagh D. Nidovirales: a new order comprising Coronaviridae and Arteriviridae. Arch Virol 142:629–633.
- Snijder EJ, Kikkert M, Fang Y. Arterivirus molecular biology and pathogenesis. J Gen Virol 94:2141–2163.
- Timoney PJ, McCollum WH. Equine viral arteritis. Vet Clin North Am Equine Pract 9:295–309.
- Timoney PJ, McCollum WH, Roberts AW, Murphy TW. Demonstration of the carrier state in naturally acquired equine arteritis virus infection in the stallion. Res Vet Sci 41:279–280.
- Miszczak F, Legrand L, Balasuriya UB, Ferry-Abitbol B, Zhang J, Hans A, Fortier G, Pronost S, Vabret A. Emergence of novel equine arteritis virus (EAV) variants during persistent infection in the stallion: origin of the 2007 French EAV outbreak was linked to an EAV strain present in the semen of a persistently infected carrier stallion. Virology 423:165–174.
- World Organization for Animal Health. Equine viral arteritis (EVA). In OIE manual of diagnostic tests and vaccines for terrestrial animals, vol 2 World Organization for Animal Health, Paris, France.
- Balasuriya UB, Leutenegger CM, Topol J B, McCollum WH, Timoney PJ, MacLachlan NJ. Detection of equine arteritis virus by real-time TaqMan reverse transcription-PCR assay. J Virol Methods 101:21–28.
- Miszczak F, Shuck KM, Lu Z, Go YY, Zhang J, Sells S, Vabret A, Pronost S, Fortier G, Timoney PJ, Balasuriya UB. Evaluation of two magnetic-bead-based viral nucleic acid purification kits and three real-time reverse transcription-PCR reagent systems in two TaqMan assays for equine arteritis virus detection in semen. J Clin Microbiol 49:3694–3696.
- Pronost S, Pitel PH, Miszczak F, Legrand L, Marcillaud-Pitel C, Hamon M, Tapprest J, Balasuriya UB, Freymuth F, Fortier G. Description of the first recorded major occurrence of equine viral arteritis in France. Equine Vet J 42:713–720.
- Berthet N, Reinhardt AK, Leclercq I, van Ooyen S, Batejat C, Dickinson P, Stamboliyska R, Old IG, Kong KA, Dacheux L, Bourhy H, Kennedy GC, Korfhage C, Cole ST, Manuguerra JC. Phi29 polymerase based random amplification of viral RNA as an alternative to random RT-PCR. BMC Mol Biol 9:77.
- Beerens N, Selisko B, Ricagno S, Imbert I, van der Zanden L, Snijder EJ, Canard B. De novo initiation of RNA synthesis by the arterivirus RNA-dependent RNA polymerase. J Virol 81:8384–8395.
- Cutler DJ, Zwick ME, Carrasquillo MM, Yohn CT, Tobin KP, Kashuk C, Mathews DJ, Shah NA, Eichler EE, Warrington JA, Chakravarti A. High-throughput variation detection and genotyping using microarrays. Genome Res 11:1913–1925.
- Gouy M, Guindon S, Gascuel O. SeaView version 4: a multiplatform graphical user interface for sequence alignment and phylogenetic tree building. Mol Biol Evol 27:221–224.
- Zhang J, Miszczak F, Pronost S, Fortier C, Balasuriya UB, Zientara S, Fortier G, Timoney PJ. Genetic variation and phylogenetic analysis of 22 French isolates of equine arteritis virus. Arch Virol 152:1977–1994.
Citations
This article has been cited 2 times.- Kourout M, Espich S, Fisher C, Tiper I, Purkayastha A, Smith S, Santana-Quintero L, Duncan R. Multiplex detection and identification of viral, bacterial, and protozoan pathogens in human blood and plasma using an expanded high-density resequencing microarray platform. Front Mol Biosci 2024;11:1419213.
- Lazić S, Lupulović D, Gaudaire D, Petrovic T, Lazić G, Hans A. Serological evidence of equine arteritis virus infection and phylogenetic analysis of viral isolates in semen of stallions from Serbia. BMC Vet Res 2017 Nov 7;13(1):316.
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