Diagnostic sensitivity and specificity of in situ hybridization and immunohistochemistry for Eastern equine encephalitis virus and West Nile virus in formalin-fixed, paraffin-embedded brain tissue of horses.
Abstract: Immunohistochemistry (IHC) and in situ hybridization (ISH) can be used either to detect or to differentiate between Eastern equine encephalitis virus (EEEV) and West Nile virus (WNV) within formalin-fixed, paraffin-embedded (FFPE) brain tissue of horses. To compare the diagnostic sensitivity and specificity of ISH and IHC, FFPE brain tissue from 20 EEEV-positive horses and 16 WNV-positive horses were tested with both EEEV and WNV oligoprobes and EEEV- and WNV-specific antibodies. Reverse transcription polymerase chain reaction (RT-PCR) for detection of EEEV and WNV was used as the gold standard to confirm infection. All horses that tested positive for EEEV by RT-PCR also tested positive by IHC and ISH, except for 1 case that was false-negative by ISH. In contrast, all horses that tested positive for WNV by RT-PCR tested negative by IHC and only 2 horses tested positive by ISH. No false-positives were detected with either method for both viruses. Both IHC and ISH are highly specific and sensitive diagnostic methods to detect EEEV in equine FFPE brain tissues, although neither appear effective for the diagnosis of WNV in equine neurologic cases.
Publication Date: 2012-03-02 PubMed ID: 22379048DOI: 10.1177/1040638711435230Google Scholar: Lookup
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- Comparative Study
- Journal Article
Summary
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The research article is about the comparison of two diagnostic methods, in situ hybridization (ISH) and immunohistochemistry (IHC), for detecting Eastern equine encephalitis virus (EEEV) and West Nile virus (WNV) in horse brain tissue.
Objective and Methodology of the Research
- The primary aim of this study was to evaluate the diagnostic sensitivity and specificity of ISH and IHC in detecting and differentiating between EEEV and WNV infections in horses using formalin-fixed, paraffin-embedded (FFPE) brain tissue samples.
- The researchers used FFPE brain tissue samples sourced from 36 horses, 20 of which had confirmed EEEV infections and 16 had confirmed WNV infections.
- The EEEV and WNV infection confirmation was based on an established gold standard technique called reverse transcription polymerase chain reaction (RT-PCR).
- Upon gathering the samples, they were then tested with both ISH and IHC methods using specific oligoprobes and antibodies for EEEV and WNV.
Research Findings
- The research findings showed that both IHC and ISH were effective in detecting EEEV in the tissue samples. All the samples which had tested positive for EEEV through RT-PCR also tested positive with both ISH and IHC, with the exception of one false-negative result with ISH.
- Unlike the EEEV results, the detection of WNV was a challenge. All samples which had tested positive for WNV through RT-PCR failed to test positive with IHC, and only 2 samples tested positive with ISH.
- The study observed that there were no false-positive detections using either method, implying a high level of specificity for both diagnostic techniques.
Conclusion
- The conclusion reached based on these research findings is that both ISH and IHC are highly effective diagnostic methods for EEEV detection in equine FFPE brain tissues in terms of both sensitivity and specificity.
- However, the same cannot be said for the diagnosis of WNV. Neither IHC nor ISH appears to be effective for the detection of WNV in equine brain tissues.
Cite This Article
APA
Pennick KE, McKnight CA, Patterson JS, Latimer KS, Maes RK, Wise AG, Kiupel M.
(2012).
Diagnostic sensitivity and specificity of in situ hybridization and immunohistochemistry for Eastern equine encephalitis virus and West Nile virus in formalin-fixed, paraffin-embedded brain tissue of horses.
J Vet Diagn Invest, 24(2), 333-338.
https://doi.org/10.1177/1040638711435230 Publication
Researcher Affiliations
- Diagnostic Center for Population and Animal Health, College of Veterinary Medicine, Michigan State University, Lansing, MI 48910, USA.
MeSH Terms
- Animals
- Brain / virology
- Encephalitis Virus, Eastern Equine / genetics
- Encephalitis Virus, Eastern Equine / isolation & purification
- Encephalomyelitis, Equine / diagnosis
- Encephalomyelitis, Equine / virology
- Horse Diseases / diagnosis
- Horse Diseases / virology
- Horses
- Immunohistochemistry / veterinary
- In Situ Hybridization / veterinary
- RNA, Viral / chemistry
- RNA, Viral / genetics
- Reverse Transcriptase Polymerase Chain Reaction / veterinary
- Sensitivity and Specificity
- West Nile Fever / diagnosis
- West Nile Fever / veterinary
- West Nile Fever / virology
- West Nile virus / genetics
- West Nile virus / isolation & purification
Citations
This article has been cited 5 times.- Matiasek K, Pfaff F, Weissenböck H, Wylezich C, Kolodziejek J, Tengstrand S, Ecke F, Nippert S, Starcky P, Litz B, Nessler J, Wohlsein P, Baumbach C, Mundhenk L, Aebischer A, Reiche S, Weidinger P, Olofsson KM, Rohdin C, Weissenbacher-Lang C, Matt J, Rosati M, Flegel T, Hörnfeldt B, Höper D, Ulrich RG, Nowotny N, Beer M, Ley C, Rubbenstroth D. Mystery of fatal 'staggering disease' unravelled: novel rustrela virus causes severe meningoencephalomyelitis in domestic cats. Nat Commun 2023 Feb 4;14(1):624.
- Cavalleri JV, Korbacska-Kutasi O, Leblond A, Paillot R, Pusterla N, Steinmann E, Tomlinson J. European College of Equine Internal Medicine consensus statement on equine flaviviridae infections in Europe. J Vet Intern Med 2022 Nov;36(6):1858-1871.
- Williams JA, Long SY, Zeng X, Kuehl K, Babka AM, Davis NM, Liu J, Trefry JC, Daye S, Facemire PR, Iversen PL, Bavari S, Pitt ML, Nasar F. Eastern equine encephalitis virus rapidly infects and disseminates in the brain and spinal cord of cynomolgus macaques following aerosol challenge. PLoS Negl Trop Dis 2022 May;16(5):e0010081.
- Meixner N, Sommer MF, Scuda N, Matiasek K, Müller M. Comparative aspects of laboratory testing for the detection of Toxoplasma gondii and its differentiation from Neospora caninum as the etiologic agent of ovine abortion. J Vet Diagn Invest 2020 Nov;32(6):898-907.
- Schwarz ER, Long MT. Comparison of West Nile Virus Disease in Humans and Horses: Exploiting Similarities for Enhancing Syndromic Surveillance. Viruses 2023 May 24;15(6).
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