Omneity® Pellets
All-In-One Vitamin & Mineral Pellet
Detection of equine arteritis virus by real-time TaqMan reverse transcription-PCR assay.
Abstract: A one-tube real-time TaqMan reverse transcription-polymerase chain reaction (RT-PCR) assay was developed for the detection of equine arteritis virus (EAV). The test was validated using the seminal plasma and nasal secretions of infected horses that were proven to contain EAV by traditional virus isolation in rabbit kidney thirteen (RK-13) cells, as well as a variety of cell culture-propagated European and North American strains of EAV. The primers and a fluorogenic TaqMan probe were designed to amplify and detect a highly conserved region of open reading frame 7 (ORF7) of EAV. The real-time TaqMan PCR assay detected EAV RNA in all samples that were confirmed to contain infectious EAV by virus isolation. The assay had an analytical sensitivity of 10 molecules of EAV RNA allowing the detection of EAV in clinical samples or tissue culture fluid (TCF) containing at least 200 viral RNA copies per ml. Thus, the one-tube real-time TaqMan RT-PCR assay provides a rapid, accurate, quantitative, convenient and high sample throughput system for diagnosis of EAV infection, in a closed-tube format that minimizes the risk of cross-contamination.
Publication Date: 2002-02-19 PubMed ID: 11849680DOI: 10.1016/s0166-0934(01)00416-5Google 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.
- Comparative Study
- Evaluation Study
- 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 article discusses the development and validation of a rapid, accurate, and highly sensitive diagnostic test for detecting Equine Arteritis Virus (EAV) using a one-tube real-time TaqMan Reverse Transcription-Polymerase Chain Reaction (RT-PCR) assay.
Development of the RT-PCR Assay
- The one-tube real-time TaqMan RT-PCR assay was designed specifically for the detection of EAV.
- The test used primers and a fluorogenic TaqMan probe to amplify and identify a highly conserved region of the open reading frame 7 (ORF7) of EAV. This area of the viral genome was targeted due to its consistent presence across diverse strains of the virus.
Validation of the Assay
- The validation process involved testing the assay on seminal plasma and nasal secretions of horses that were known to be infected with EAV. These samples contained active, infectious versions of the virus, as confirmed by traditional virus isolation methods in rabbit kidney thirteen (RK-13) cells.
- The test was also applied on a range of cell culture-propagated strains of EAV from Europe and North America to ensure its efficacy across diverse EAV strains.
- The RT-PCR assay was able to identify EAV RNA in all samples that were confirmed to contain the virus through traditional isolation methods, effectively demonstrating the assay’s high degree of accuracy.
Characteristics and Advantages of the Assay
- The RT-PCR assay was found to possess high analytical sensitivity, with a capacity to detect as few as 10 molecules of EAV RNA. This allows the diagnosis of EAV infection in clinical samples or tissue culture fluid (TCF) containing at least 200 viral RNA copies per milliliter.
- The assay offers a quick, precise, and quantifiable system for diagnosing EAV infection. This improves upon current systems by offering a high sample throughput, meaning it can process numerous samples efficiently.
- Furthermore, the one-tube, closed-tube format of the assay was designed to minimize the risk of cross-contamination, making it a more secure and reliable method for virus detection.
Cite This Article
APA
Balasuriya UB, Leutenegger CM, Topol JB, McCollum WH, Timoney PJ, MacLachlan NJ.
(2002).
Detection of equine arteritis virus by real-time TaqMan reverse transcription-PCR assay.
J Virol Methods, 101(1-2), 21-28.
https://doi.org/10.1016/s0166-0934(01)00416-5 Publication
Researcher Affiliations
- Bernard and Gloria Salick Equine Viral Disease Laboratory, Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, One Shields Avenue, University of California, Davis, CA 95616, USA. ubbalasuriya@ucdavis.edu
MeSH Terms
- Animals
- Arterivirus Infections / diagnosis
- Arterivirus Infections / veterinary
- Cell Line
- Conserved Sequence
- Equartevirus / genetics
- Equartevirus / isolation & purification
- Europe
- Horse Diseases / virology
- Horses
- Male
- Nasal Mucosa / metabolism
- Nasal Mucosa / virology
- North America
- Open Reading Frames
- RNA, Viral / analysis
- Rabbits
- Reproducibility of Results
- Reverse Transcriptase Polymerase Chain Reaction / veterinary
- Semen / virology
- Sensitivity and Specificity
- Taq Polymerase / chemistry
- Time Factors
Citations
This article has been cited 27 times.- Bhat S, Karunakaran S, Frossard JP, Choudhury B, Steinbach F. Genetic characterization of equine arteritis virus associated with outbreaks in the UK, 2019. J Gen Virol 2025 Dec;106(12).
- Knox A, Beddoe T. Isothermal Nucleic Acid Amplification Technologies for the Detection of Equine Viral Pathogens. Animals (Basel) 2021 Jul 20;11(7).
- Wang CYT, Ballard E, Llewellyn S, Marquart L, Bousema T, McCarthy JS, Collins KA. Assays for quantification of male and female gametocytes in human blood by qRT-PCR in the absence of pure sex-specific gametocyte standards. Malar J 2020 Jun 23;19(1):218.
- . Opinion of the Scientific Panel on Animal Health and Welfare (AHAW) on a request from the Commission related to the probability of transmission of Porcine Reproductive and Respiratory Syndrome virus (PRRSv) to naive pigs via fresh meat. EFSA J 2005 Aug;3(8):239.
- Grandolfo E, Parisi A, Ricci A, Lorusso E, de Siena R, Trotta A, Buonavoglia D, Martella V, Corrente M. High mortality in foals associated with Salmonella enterica subsp. enterica Abortusequi infection in Italy. J Vet Diagn Invest 2018 May;30(3):483-485.
- 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.
- Björnsdóttir S, Harris SR, Svansson V, Gunnarsson E, Sigurðardóttir ÓG, Gammeljord K, Steward KF, Newton JR, Robinson C, Charbonneau ARL, Parkhill J, Holden MTG, Waller AS. Genomic Dissection of an Icelandic Epidemic of Respiratory Disease in Horses and Associated Zoonotic Cases. mBio 2017 Aug 1;8(4).
- 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).
- Nagy A, Vitásková E, Černíková L, Křivda V, Jiřincová H, Sedlák K, Horníčková J, Havlíčková M. Evaluation of TaqMan qPCR System Integrating Two Identically Labelled Hydrolysis Probes in Single Assay. Sci Rep 2017 Jan 25;7:41392.
- Nagy A, Černíková L, Vitásková E, Křivda V, Dán Á, Dirbáková Z, Jiřincová H, Procházka B, Sedlák K, Havlíčková M. MeltMan: Optimization, Evaluation, and Universal Application of a qPCR System Integrating the TaqMan qPCR and Melting Analysis into a Single Assay. PLoS One 2016;11(3):e0151204.
- Back H, Penell J, Pringle J, Isaksson M, Ronéus N, Treiberg Berndtsson L, Ståhl K. A longitudinal study of poor performance and subclinical respiratory viral activity in Standardbred trotters. Vet Rec Open 2015;2(1):e000107.
- Nemoto M, Bannai H, Tsujimura K, Kobayashi M, Kikuchi T, Yamanaka T, Kondo T. Getah Virus Infection among Racehorses, Japan, 2014. Emerg Infect Dis 2015 May;21(5):883-5.
- 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.
- Verhagen LM, Gómez-Castellano K, Snelders E, Rivera-Olivero I, Pocaterra L, Melchers WJ, de Waard JH, Hermans PW. Respiratory infections in Eñepa Amerindians are related to malnutrition and Streptococcus pneumoniae carriage. J Infect 2013 Oct;67(4):273-81.
- Lu Z, Timoney PJ, White J, Balasuriya UB. Development of one-step TaqMan® real-time reverse transcription-PCR and conventional reverse transcription-PCR assays for the detection of equine rhinitis A and B viruses. BMC Vet Res 2012 Jul 25;8:120.
- Zhang J, Go YY, Huang CM, Meade BJ, Lu Z, Snijder EJ, Timoney PJ, Balasuriya UB. Development and characterization of an infectious cDNA clone of the modified live virus vaccine strain of equine arteritis virus. Clin Vaccine Immunol 2012 Aug;19(8):1312-21.
- Pratelli A. The evolutionary processes of canine coronaviruses. Adv Virol 2011;2011:562831.
- Sanghavi SK, Bullotta A, Husain S, Rinaldo CR. Clinical evaluation of multiplex real-time PCR panels for rapid detection of respiratory viral infections. J Med Virol 2012 Jan;84(1):162-9.
- 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 2011 Oct;49(10):3694-6.
- Ding M, Bullotta A, Caruso L, Gupta P, Rinaldo CR, Chen Y. An optimized sensitive method for quantitation of DNA/RNA viruses in heparinized and cryopreserved plasma. J Virol Methods 2011 Sep;176(1-2):1-8.
- Go YY, Zhang J, Timoney PJ, Cook RF, Horohov DW, Balasuriya UB. Complex interactions between the major and minor envelope proteins of equine arteritis virus determine its tropism for equine CD3+ T lymphocytes and CD14+ monocytes. J Virol 2010 May;84(10):4898-911.
- Zhang J, Stein DA, Timoney PJ, Balasuriya UB. Curing of HeLa cells persistently infected with equine arteritis virus by a peptide-conjugated morpholino oligomer. Virus Res 2010 Jun;150(1-2):138-42.
- Dare R, Sanghavi S, Bullotta A, Keightley MC, George KS, Wadowsky RM, Paterson DL, McCurry KR, Reinhart TA, Husain S, Rinaldo CR. Diagnosis of human metapneumovirus infection in immunosuppressed lung transplant recipients and children evaluated for pertussis. J Clin Microbiol 2007 Feb;45(2):548-52.
- Revilla-Fernández S, Wallner B, Truschner K, Benczak A, Brem G, Schmoll F, Mueller M, Steinborn R. The use of endogenous and exogenous reference RNAs for qualitative and quantitative detection of PRRSV in porcine semen. J Virol Methods 2005 Jun;126(1-2):21-30.
- Burnouf T, Griffiths E, Padilla A, Seddik S, Stephano MA, Gutiérrez JM. Assessment of the viral safety of antivenoms fractionated from equine plasma. Biologicals 2004 Sep;32(3):115-28.
- Decaro N, Pratelli A, Campolo M, Elia G, Martella V, Tempesta M, Buonavoglia C. Quantitation of canine coronavirus RNA in the faeces of dogs by TaqMan RT-PCR. J Virol Methods 2004 Aug;119(2):145-50.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
