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.
Abstract: This study showed that under specifically defined conditions with respect to nucleic acid extraction method and testing reagents, a previously described real-time reverse transcription-PCR (rRT-PCR) assay (T1 assay) provides sensitivity equal to or higher than that of virus isolation for the detection of equine arteritis virus in semen.
Publication Date: 2011-08-10 PubMed ID: 21832018PubMed Central: PMC3187312DOI: 10.1128/JCM.01187-11Google Scholar: Lookup
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- Evaluation Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research paper compares two kits used for viral nucleic acid purification and three systems for real-time reverse transcription-PCR — a method used to detect specific genetic material in a DNA sample — in their effectiveness for identifying equine arteritis virus in horse semen. The findings suggest that under certain conditions, one method called rRT-PCR can detect the virus as effectively, or more so, than virus isolation.
Research Methods and Materials
- The research made use of two different kits specially designed for the purification of viral nucleic acid from horse semen. These kits use magnetic beads as the primary method of isolating the nucleic acids.
- Three different real-time reverse transcription-PCR (rRT-PCR) systems were also tested. rRT-PCR is a technique used to copy and amplify specific segments of DNA which allows for the detection of specific genetic sequences, like those of viruses. Here, it was used to detect the presence of equine arteritis virus.
- The study involved two specific TaqMan assays. TaqMan is a technology used in real-time PCR to detect and quantify DNA. The two assays used here were designed to identify the presence of equine arteritis virus.
Results and Findings
- The researchers reported that, under specific conditions, the use of an rRT-PCR assay, referred to as the T1 assay, showed equal or higher sensitivity than virus isolation in detecting equine arteritis virus in horse semen.
- This finding suggests the potential effectiveness of the T1 assay as a faster, less resource-intensive alternative to the traditional method of virus isolation.
Implications and Relevance
- The study provided insight into the most efficient methods for detecting equine arteritis virus, which is important for controlling and preventing outbreaks in equine populations.
- Such findings may lead to more effective and efficient diagnostic practices, improving veterinary healthcare for horses and other equine species.
Cite This Article
APA
Miszczak F, Shuck KM, Lu Z, Go YY, Zhang J, Sells S, Vabret A, Pronost S, Fortier G, Timoney PJ, Balasuriya UB.
(2011).
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(10), 3694-3696.
https://doi.org/10.1128/JCM.01187-11 Publication
Researcher Affiliations
- Frank Duncombe Laboratory, Animal Health Department, IFR146 ICORE, 14053 Caen, France.
MeSH Terms
- Animals
- Arterivirus Infections / diagnosis
- Arterivirus Infections / veterinary
- Arterivirus Infections / virology
- Equartevirus / genetics
- Equartevirus / isolation & purification
- Horse Diseases / diagnosis
- Horse Diseases / virology
- Horses
- Immunomagnetic Separation / methods
- RNA, Viral / genetics
- RNA, Viral / isolation & purification
- Reverse Transcriptase Polymerase Chain Reaction / methods
- Semen / virology
- Sensitivity and Specificity
- Veterinary Medicine / methods
- Virology / methods
References
This article includes 18 references
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Citations
This article has been cited 5 times.- Nam B, Mekuria Z, Carossino M, Li G, Zheng Y, Zhang J, Cook RF, Shuck KM, Campos JR, Squires EL, Troedsson MHT, Timoney PJ, Balasuriya UBR. Intrahost Selection Pressure Drives Equine Arteritis Virus Evolution during Persistent Infection in the Stallion Reproductive Tract. J Virol 2019 Jun 15;93(12).
- Carossino M, Loynachan AT, Canisso IF, Cook RF, Campos JR, Nam B, Go YY, Squires EL, Troedsson MHT, Swerczek T, Del Piero F, Bailey E, Timoney PJ, Balasuriya UBR. Equine Arteritis Virus Has Specific Tropism for Stromal Cells and CD8(+) T and CD21(+) B Lymphocytes but Not for Glandular Epithelium at the Primary Site of Persistent Infection in the Stallion Reproductive Tract. J Virol 2017 Jul 1;91(13).
- Hans A, Gaudaire D, Manuguerra JC, Leon A, Gessain A, Laugier C, Berthet N, Zientara S. Combination of an unbiased amplification method and a resequencing microarray for detecting and genotyping equine arteritis virus. J Clin Microbiol 2015 Jan;53(1):287-91.
- Balasuriya UB, Go YY, MacLachlan NJ. Equine arteritis virus. Vet Microbiol 2013 Nov 29;167(1-2):93-122.
- Sochor J, Babula P, Adam V, Krska B, Kizek R. Sharka: the past, the present and the future. Viruses 2012 Nov 7;4(11):2853-901.
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