15 results on '"Shuck, Kathleen M."'
Search Results
2. Detection of equine arteritis virus by two chromogenic RNA in situ hybridization assays (conventional and RNAscope®) and assessment of their performance in tissues from aborted equine fetuses
- Author
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Carossino, Mariano, Loynachan, Alan T., James MacLachlan, N., Drew, Clifton, Shuck, Kathleen M., Timoney, Peter J., Del Piero, Fabio, and Balasuriya, Udeni B. R.
- Published
- 2016
- Full Text
- View/download PDF
3. Response of Stallions to Primary Immunization with a Modified Live Equine Viral Arteritis Vaccine
- Author
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Summers-Lawyer, Amy K., Go, Yun Young, Lu, Zhengchun, Timoney, Peter J., McCue, Patrick M., Zhang, Jianqiang, Shuck, Kathleen M., and Bruemmer, Jason
- Published
- 2011
- Full Text
- View/download PDF
4. Allelic Variation in CXCL16 Determines CD3+ T Lymphocyte Susceptibility to Equine Arteritis Virus Infection and Establishment of Long-Term Carrier State in the Stallion.
- Author
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Sarkar, Sanjay, Bailey, Ernest, Go, Yun Young, Cook, R. Frank, Kalbfleisch, Ted, Eberth, John, Chelvarajan, R. Lakshman, Shuck, Kathleen M., Artiushin, Sergey, Timoney, Peter J., and Balasuriya, Udeni B. R.
- Subjects
HORSE diseases ,ARTERITIS ,CD3 antigen ,LYMPHOCYTES ,VIRUS diseases ,CHEMOKINES ,SINGLE nucleotide polymorphisms - Abstract
Equine arteritis virus (EAV) is the causative agent of equine viral arteritis (EVA), a respiratory, systemic, and reproductive disease of horses and other equid species. Following natural infection, 10–70% of the infected stallions can become persistently infected and continue to shed EAV in their semen for periods ranging from several months to life. Recently, we reported that some stallions possess a subpopulation(s) of CD3
+ T lymphocytes that are susceptible to in vitro EAV infection and that this phenotypic trait is associated with long-term carrier status following exposure to the virus. In contrast, stallions not possessing the CD3+ T lymphocyte susceptible phenotype are at less risk of becoming long-term virus carriers. A genome wide association study (GWAS) using the Illumina Equine SNP50 chip revealed that the ability of EAV to infect CD3+ T lymphocytes and establish long-term carrier status in stallions correlated with a region within equine chromosome 11. Here we identified the gene and mutations responsible for these phenotypes. Specifically, the work implicated three allelic variants of the equine orthologue of CXCL16 (EqCXCL16) that differ by four non-synonymous nucleotide substitutions (XM_00154756; c.715 A → T, c.801 G → C, c.804 T → A/G, c.810 G → A) within exon 1. This resulted in four amino acid changes with EqCXCL16S (XP_001504806.1) having Phe, His, Ile and Lys as compared to EqCXL16R having Tyr, Asp, Phe, and Glu at 40, 49, 50, and 52, respectively. Two alleles (EqCXCL16Sa, EqCXCL16Sb) encoded identical protein products that correlated strongly with long-term EAV persistence in stallions (P<0.000001) and are required for in vitro CD3+ T lymphocyte susceptibility to EAV infection. The third (EqCXCL16R) was associated with in vitro CD3+ T lymphocyte resistance to EAV infection and a significantly lower probability for establishment of the long-term carrier state (viral persistence) in the male reproductive tract. EqCXCL16Sa and EqCXCL16Sb exert a dominant mode of inheritance. Most importantly, the protein isoform EqCXCL16S but not EqCXCL16R can function as an EAV cellular receptor. Although both molecules have equal chemoattractant potential, EqCXCL16S has significantly higher scavenger receptor and adhesion properties compared to EqCXCL16R. [ABSTRACT FROM AUTHOR]- Published
- 2016
- Full Text
- View/download PDF
5. The Replication of Poliovirus in Primary Kidney Cell Cultures of Thirteen-Lined Ground Squirrels.
- Author
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McKenna, John M., Shuck, Kathleen M., and Cashon, Gary W.
- Published
- 1971
- Full Text
- View/download PDF
6. Intra-host Selection Pressure Drives Equine Arteritis Virus Evolution during Persistent Infection in the Stallion Reproductive Tract.
- Author
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Nam, Bora, Mekuria, Zelalem, Carossino, Mariano, Ganwu Li, Ying Zheng, Jianqiang Zhang, Cook, R. Frank, Shuck, Kathleen M., Campos, Juliana R., Squires, Edward L., Troedsson, Mats H. T., Timoney, Peter J., and Balasuriya, Udeni B. R.
- Subjects
- *
GENITALIA , *VIRAL genomes , *STALLIONS , *ARTERITIS , *VAS deferens , *FEMALE reproductive organs , *HORSE diseases - Abstract
Equine arteritis virus (EAV) is the causative agent of equine viral arteritis (EVA), a reproductive and respiratory disease of horses. Following natural infection, 10-70% of infected stallions can become carriers of EAV and continue to shed virus in the semen. In this study, sequential viruses isolated from nasal secretions, buffy coat cells and semen of seven experimentally infected and two naturally infected EAV carrier stallions were deep sequenced to elucidate the intra-host microevolutionary process after a single transmission event. Analysis of variants from nasal secretions and buffy coat cells lacked extensive positive selection; however, characteristics of the mutant spectra were different in the two sample types. By contrast, the initial semen virus populations during acute infection have undergone a selective bottleneck as reflected by the reduction in population size and diversifying selection at multiple sites in the viral genome. Furthermore, during persistent infection period, extensive genome-wide purifying selection shaped variant diversity in stallion reproductive tract. Overall, the non- stochastic nature of EAV evolution during persistent infection was driven by active intra-host selection pressure. Among the open reading frames within the viral genome, ORF3, ORF5 and the nsp2 encoding region of ORF1a accumulated the majority of nucleotide substitutions during persistence, with ORF3 and ORF5 having the highest intra-host evolutionary rates. The findings presented here provide a novel insight into the evolutionary mechanisms of EAV, identified critical regions of the viral genome likely associated with the establishment and maintenance of persistent infection in the stallion reproductive tract. Importance EAV can persist in the reproductive tract of infected stallions and, consequently, long- term carrier stallions constitute its sole natural reservoir. Previous studies demonstrated that the ampullae of the vas deferens is the primary site of viral persistence in the stallion reproductive tract, and is associated with a significant inflammatory response that is unable to clear the infection. This is the first study that describes EAV full-length genomic evolution during acute and long-term persistent infection in the stallion reproductive tract using next-generation sequencing and contemporary sequence analysis techniques. The data provide novel insight into the intra-host evolution of EAV during acute and persistent infection and demonstrates that persistent infection is characterized by extensive genome-wide purifying selection and a non-stochastic evolutionary pattern mediated by intra-host selective pressure, with important nucleotide substitutions occurring in ORFs 1a (region encoding nsp2), 3 and 5. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
7. Downregulation of MicroRNA eca-mir-128 in Seminal Exosomes and Enhanced Expression of CXCL16 in the Stallion Reproductive Tract Are Associated with Long-Term Persistence of Equine Arteritis Virus.
- Author
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Carossino, Mariano, Dini, Pouya, Kalbfleisch, Theodore S., Loynachan, Alan T., Canisso, Igor F., Shuck, Kathleen M., Timoney, Peter J., Cook, R. Frank, and Balasuriya, Udeni B. R.
- Subjects
- *
ARTERITIS , *STALLIONS , *SEMEN , *RIBONUCLEASES , *IMMUNOGLOBULINS - Abstract
Equine arteritis virus (EAV) can establish long-term persistent infection in the reproductive tract of stallions and is shed in the semen. Previous studies showed that long-term persistence is associated with a specific allele of the CXCL16 gene (CXCL16S) and that persistent infection is maintained despite the presence of a local inflammatory and humoral and mucosal antibody responses. In this study, we demonstrated that equine seminal exosomes (SEs) are enriched in a small subset of microRNAs (miRNAs). Most importantly, we demonstrated that long-term EAV persistence is associated with the downregulation of an SE-associated miRNA (eca-mir- 128) and with an enhanced expression of CXCL16 in the reproductive tract, a putative target of eca-mir-128. The findings presented here suggest that SE eca-mir-128 is implicated in the regulation of the CXCL16/CXCR6 axis in the reproductive tract of persistently infected stallions, a chemokine axis strongly implicated in EAV persistence. This is a novel finding and warrants further investigation to identify its specific mechanism in modulating the CXCL16/CXCR6 axis in the reproductive tract of the EAV long-term carrier stallion. IMPORTANCE Equine arteritis virus (EAV) has the ability to establish long-term persistent infection in the stallion reproductive tract and to be shed in semen, which jeopardizes its worldwide control. Currently, the molecular mechanisms of viral persistence are being unraveled and these are essential for the development of effective therapeutics to eliminate persistent infection. Recently, it has been determined that long-term persistence is associated with a specific allele of the CXCL16 gene (CXCL16S) and is maintained despite induction of local inflammatory, humoral and mucosal antibody responses. This study demonstrated that long-term persistence is associated with the downregulation of seminal exosome miRNA eca-mir-128 and enhanced expression of its putative target, CXCL16, in the reproductive tract. For the first time, this study suggests complex interactions between eca-mir-128 and cellular elements at the site of EAV persistence and implicates this miRNA in the regulation of the CXCL16/CXCR6 axis in the reproductive tract during long-term persistence. [ABSTRACT FROM AUTHOR]
- Published
- 2018
- Full Text
- View/download PDF
8. Evidence that In Vitro Susceptibility of CD3+ T Lymphocytes to Equine Arteritis Virus Infection Reflects Genetic Predisposition of Naturally Infected Stallions To Become Carriers of the Virus.
- Author
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Go, Yun Young, Bailey, Ernest, Timoney, Peter J., Shuck, Kathleen M., and Balasuriya, Udeni B. R.
- Subjects
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T cells , *ARTERITIS , *VIRUS diseases , *DISEASE susceptibility , *STALLIONS , *PHENOTYPES , *DISEASES - Abstract
We investigated the correlation between in vitro susceptibility of CD3+ T lymphocytes to equine arteritis virus (EAV) infection and establishment of persistent infection among 14 stallions following natural infections. The data showed that carrier stallions with a CD3+ T lymphocyte susceptibility phenotype to in vitro EAV infection may be at higher risk of becoming carriers than those that lack this phenotype (P = 0.0002). [ABSTRACT FROM AUTHOR]
- Published
- 2012
- Full Text
- View/download PDF
9. Allelic Variation in CXCL16 Determines CD3+ T Lymphocyte Susceptibility to Equine Arteritis Virus Infection and Establishment of Long-Term Carrier State in the Stallion.
- Author
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Sarkar S, Bailey E, Go YY, Cook RF, Kalbfleisch T, Eberth J, Chelvarajan RL, Shuck KM, Artiushin S, Timoney PJ, and Balasuriya UB
- Subjects
- Alleles, Amino Acid Sequence genetics, Animals, Arterivirus Infections veterinary, Arterivirus Infections virology, CD3 Complex genetics, CD3 Complex immunology, Equartevirus pathogenicity, Genetic Predisposition to Disease, Genome-Wide Association Study, Horse Diseases virology, Horses genetics, Horses virology, Male, Phylogeny, Semen metabolism, T-Lymphocytes immunology, T-Lymphocytes pathology, Arterivirus Infections genetics, Chemokines, CXC genetics, Equartevirus genetics, Horse Diseases genetics
- Abstract
Equine arteritis virus (EAV) is the causative agent of equine viral arteritis (EVA), a respiratory, systemic, and reproductive disease of horses and other equid species. Following natural infection, 10-70% of the infected stallions can become persistently infected and continue to shed EAV in their semen for periods ranging from several months to life. Recently, we reported that some stallions possess a subpopulation(s) of CD3+ T lymphocytes that are susceptible to in vitro EAV infection and that this phenotypic trait is associated with long-term carrier status following exposure to the virus. In contrast, stallions not possessing the CD3+ T lymphocyte susceptible phenotype are at less risk of becoming long-term virus carriers. A genome wide association study (GWAS) using the Illumina Equine SNP50 chip revealed that the ability of EAV to infect CD3+ T lymphocytes and establish long-term carrier status in stallions correlated with a region within equine chromosome 11. Here we identified the gene and mutations responsible for these phenotypes. Specifically, the work implicated three allelic variants of the equine orthologue of CXCL16 (EqCXCL16) that differ by four non-synonymous nucleotide substitutions (XM_00154756; c.715 A → T, c.801 G → C, c.804 T → A/G, c.810 G → A) within exon 1. This resulted in four amino acid changes with EqCXCL16S (XP_001504806.1) having Phe, His, Ile and Lys as compared to EqCXL16R having Tyr, Asp, Phe, and Glu at 40, 49, 50, and 52, respectively. Two alleles (EqCXCL16Sa, EqCXCL16Sb) encoded identical protein products that correlated strongly with long-term EAV persistence in stallions (P<0.000001) and are required for in vitro CD3+ T lymphocyte susceptibility to EAV infection. The third (EqCXCL16R) was associated with in vitro CD3+ T lymphocyte resistance to EAV infection and a significantly lower probability for establishment of the long-term carrier state (viral persistence) in the male reproductive tract. EqCXCL16Sa and EqCXCL16Sb exert a dominant mode of inheritance. Most importantly, the protein isoform EqCXCL16S but not EqCXCL16R can function as an EAV cellular receptor. Although both molecules have equal chemoattractant potential, EqCXCL16S has significantly higher scavenger receptor and adhesion properties compared to EqCXCL16R., Competing Interests: The authors have declared that no competing interests exist.
- Published
- 2016
- Full Text
- View/download PDF
10. Complete Genome Sequence of Noncytopathic Bovine Viral Diarrhea Virus 1 Contaminating a High-Passage RK-13 Cell Line.
- Author
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Nam B, Li G, Zheng Y, Zhang J, Shuck KM, Timoney PJ, and Balasuriya UB
- Abstract
A high-passage rabbit kidney RK-13 cell line (HP-RK-13[KY], originally derived from the ATCC CCL-37 cell line) used in certain laboratories worldwide is contaminated with noncytopathic bovine viral diarrhea virus (ncpBVDV). On complete genome sequence analysis, the virus strain was found to belong to BVDV group 1b., (Copyright © 2015 Nam et al.)
- Published
- 2015
- Full Text
- View/download PDF
11. Evidence that in vitro susceptibility of CD3+ T lymphocytes to equine arteritis virus infection reflects genetic predisposition of naturally infected stallions to become carriers of the virus.
- Author
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Go YY, Bailey E, Timoney PJ, Shuck KM, and Balasuriya UB
- Subjects
- Animals, Arterivirus Infections metabolism, Arterivirus Infections transmission, Carrier State veterinary, Genetic Predisposition to Disease, Haplotypes, Horses, In Vitro Techniques, Male, Microscopy, Fluorescence methods, Phenotype, Risk, Arterivirus Infections virology, CD3 Complex biosynthesis, Equartevirus metabolism, Horse Diseases virology, T-Lymphocytes virology
- Abstract
We investigated the correlation between in vitro susceptibility of CD3(+) T lymphocytes to equine arteritis virus (EAV) infection and establishment of persistent infection among 14 stallions following natural infections. The data showed that carrier stallions with a CD3(+) T lymphocyte susceptibility phenotype to in vitro EAV infection may be at higher risk of becoming carriers than those that lack this phenotype (P = 0.0002).
- Published
- 2012
- Full Text
- View/download PDF
12. 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.
- Author
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Miszczak F, Shuck KM, Lu Z, Go YY, Zhang J, Sells S, Vabret A, Pronost S, Fortier G, Timoney PJ, and Balasuriya UB
- Subjects
- Animals, Arterivirus Infections diagnosis, Arterivirus Infections virology, Equartevirus genetics, Horse Diseases virology, Horses, RNA, Viral genetics, Sensitivity and Specificity, Veterinary Medicine methods, Virology methods, Arterivirus Infections veterinary, Equartevirus isolation & purification, Horse Diseases diagnosis, Immunomagnetic Separation methods, RNA, Viral isolation & purification, Reverse Transcriptase Polymerase Chain Reaction methods, Semen virology
- 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.
- Published
- 2011
- Full Text
- View/download PDF
13. Molecular epidemiology and genetic characterization of equine arteritis virus isolates associated with the 2006-2007 multi-state disease occurrence in the USA.
- Author
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Zhang J, Timoney PJ, Shuck KM, Seoul G, Go YY, Lu Z, Powell DG, Meade BJ, and Balasuriya UB
- Subjects
- Amino Acid Sequence, Animals, Arterivirus Infections epidemiology, Arterivirus Infections virology, Base Sequence, DNA, Viral genetics, Disease Outbreaks veterinary, Equartevirus immunology, Equartevirus isolation & purification, Evolution, Molecular, Female, Horses, Male, Molecular Epidemiology, Molecular Sequence Data, Neutralization Tests, Phenotype, Phylogeny, Pregnancy, Sequence Homology, Amino Acid, Sequence Homology, Nucleic Acid, Time Factors, United States epidemiology, Arterivirus Infections veterinary, Equartevirus genetics, Horse Diseases epidemiology, Horse Diseases virology
- Abstract
In 2006-2007, equine viral arteritis (EVA) was confirmed for the first time in Quarter Horses in multiple states in the USA. The entire genome of an equine arteritis virus (EAV) isolate from the index premises in New Mexico was 12 731 nt in length and possessed a previously unrecorded unique 15 nt insertion in the nsp2-coding region in ORF1a and a 12 nt insertion in ORF3. Sequence analysis of additional isolates made during this disease occurrence revealed that all isolates from New Mexico, Utah, Kansas, Oklahoma and Idaho had 98.6-100.0 % (nsp2) and 97.8-100 % (ORF3) nucleotide identity and contained the unique insertions in nsp2 and ORF3, indicating that the EVA outbreaks in these states probably originated from the same strain of EAV. Sequence and phylogenetic analysis of several EAV isolates made following an EVA outbreak on another Quarter Horse farm in New Mexico in 2005 provided evidence that this outbreak may well have been the source of virus for the 2006-2007 occurrence of the disease. A virus isolate from an aborted fetus in Utah was shown to have a distinct neutralization phenotype compared with other isolates associated with the 2006-2007 EVA occurrence. Full-length genomic sequence analysis of 18 sequential isolates of EAV made from eight carrier stallions established that the virus evolved genetically during persistent infection, and the rate of genetic change varied between individual animals and the period of virus shedding.
- Published
- 2010
- Full Text
- View/download PDF
14. Comparison of two real-time reverse transcription polymerase chain reaction assays for the detection of Equine arteritis virus nucleic acid in equine semen and tissue culture fluid.
- Author
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Lu Z, Branscum AJ, Shuck KM, Zhang J, Dubovi EJ, Timoney PJ, and Balasuriya UB
- Subjects
- Animals, Arterivirus Infections diagnosis, Arterivirus Infections virology, Equartevirus genetics, Horse Diseases diagnosis, Horses, Male, RNA, Viral chemistry, RNA, Viral genetics, Reproducibility of Results, Reverse Transcriptase Polymerase Chain Reaction methods, Sensitivity and Specificity, Arterivirus Infections veterinary, Equartevirus isolation & purification, Horse Diseases virology, RNA, Viral analysis, Reverse Transcriptase Polymerase Chain Reaction veterinary, Semen virology
- Abstract
Two previously developed TaqMan fluorogenic probe-based 1-tube real-time reverse transcription polymerase chain reaction (real-time RT-PCR) assays (T1 and T2) were compared and validated for the detection of Equine arteritis virus (EAV) nucleic acid in equine semen and tissue culture fluid (TCF). The specificity and sensitivity of these 2 molecular-based assays were compared to traditional virus isolation (VI) in cell culture. The T1 real-time RT-PCR had a higher sensitivity (93.4%) than the T2 real-time RT-PCR (42.6%) for detection of EAV RNA in semen. However, the T1 real-time RT-PCR was less sensitive (93.4%) than the World Organization for Animal Health (OIE)-prescribed VI test (gold standard). The sensitivity of both PCR assays was high (100.0% [T1] and 95.2% [T2]) for detecting EAV RNA in TCF. In light of the discrepancy in sensitivity between either real-time RT-PCR assay and VI, semen that is negative for EAV nucleic acid by real-time RT-PCR that is from an EAV-seropositive stallion should be confirmed free of virus by VI. Similarly, the presence of EAV in TCF samples that are VI-positive but real-time RT-PCR-negative should be confirmed in a 1-way neutralization test using anti-EAV equine serum or by fluorescent antibody test using monoclonal antibodies to EAV. If the viral isolate is not identified as EAV, such samples should be tested for other equine viral pathogens. The results of this study underscore the importance of comparative evaluation and validation of real-time RT-PCR assays prior to their recommended use in a diagnostic setting for the detection and identification of specific infectious agents.
- Published
- 2008
- Full Text
- View/download PDF
15. Development of a fluorescent-microsphere immunoassay for detection of antibodies specific to equine arteritis virus and comparison with the virus neutralization test.
- Author
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Go YY, Wong SJ, Branscum AJ, Demarest VL, Shuck KM, Vickers ML, Zhang J, McCollum WH, Timoney PJ, and Balasuriya UB
- Subjects
- Amino Acid Sequence, Animals, Antibodies, Viral immunology, Arterivirus Infections immunology, Arterivirus Infections virology, Equartevirus isolation & purification, Fluorescent Antibody Technique economics, Horses, Immunoassay economics, Microspheres, Molecular Sequence Data, Neutralization Tests methods, Polymerase Chain Reaction methods, Recombinant Proteins genetics, Sensitivity and Specificity, Viral Structural Proteins genetics, Viral Structural Proteins immunology, Antibodies, Viral analysis, Arterivirus Infections veterinary, Equartevirus immunology, Fluorescent Antibody Technique methods, Horse Diseases immunology, Horse Diseases virology, Immunoassay methods
- Abstract
The development and validation of a microsphere immunoassay (MIA) to detect equine antibodies to the major structural proteins of equine arteritis virus (EAV) are described. The assay development process was based on the cloning and expression of genes for full-length individual major structural proteins (GP5 amino acids 1 to 255 [GP5(1-255)], M(1-162), and N(1-110)), as well as partial sequences of these structural proteins (GP5(1-116), GP5(75-112), GP5(55-98), M(88-162), and N(1-69)) that constituted putative antigenic regions. Purified recombinant viral proteins expressed in Escherichia coli were covalently bound to fluorescent polystyrene microspheres and analyzed with the Luminex xMap 100 instrument. Of the eight recombinant proteins, the highest concordance with the virus neutralization test (VNT) results was obtained with the partial GP5(55-98) protein. The MIA was validated by testing a total of 2,500 equine serum samples previously characterized by the VNT. With the use of an optimal median fluorescence intensity cutoff value of 992, the sensitivity and specificity of the assay were 92.6% and 92.9%, respectively. The GP5(55-98) MIA and VNT outcomes correlated significantly (r = 0.84; P < 0.0001). Although the GP5(55-98) MIA is less sensitive than the standard VNT, it has the potential to provide a rapid, convenient, and more economical test for screening equine sera for the presence of antibodies to EAV, with the VNT then being used as a confirmatory assay.
- Published
- 2008
- Full Text
- View/download PDF
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