The identification of equid herpesvirus 1 in paraffin-embedded tissues from aborted fetuses by polymerase chain reaction and immunohistochemistry.
Abstract: Paraffin-embedded organ samples from 28 aborted fetuses and three foals, partly archival and partly sampled in 1991, were examined by polymerase chain reaction (PCR) and immunohistochemistry for the presence of DNA and antigens, respectively, specific for equine herpesvirus 1 (EHV-1). Virologic examination had been performed on 23 of the aborted fetuses. DNA fragments specific for EHV-1 were identified by PCR, and EHV-1 antigens were identified in situ by immunohistochemistry, with an agreement between the methods of 94% (kappa = 0.85). Compared with virus isolation, PCR agreement was 87% (kappa 0.69), and IH agreement was 82% (kappa = 0.47). These results showed that there was moderate to almost perfect agreement among the different methods and that PCR and immunohistochemistry are powerful tools for the identification of EHV-1 in paraffin-embedded tissues. The techniques give more rapid results than virus isolation and also detect inactivated virus, which are not identified by standard virus isolation. These techniques also make retrospective studies possible.
Publication Date: 1993-04-01 PubMed ID: 8389598DOI: 10.1177/104063879300500206Google Scholar: Lookup
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- Journal Article
Summary
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This study looks at the use of polymerase chain reaction (PCR) and immunohistochemistry to identify equine herpesvirus 1 (EHV-1) in samples from aborted horse fetuses, and found these methods very reliable even with paraffin-embedded tissues.
Understanding the Research
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The research study looked at organ samples from 28 aborted horse fetuses and three foals.
Methodology
- The researchers used two primary techniques to analyze the samples: a polymerase chain reaction (PCR) and immunohistochemistry.
- PCR allows for the amplification of specific DNA sequences for their detection. In this study, it was used to identify DNA fragments specific for EHV-1.
- Immunohistochemistry is a laboratory process used to visually detect antigens in tissue samples via the use of antibodies. In this case, it was used to identify EHV-1 antigens.
- 23 of the aborted fetus samples had also been tested with virology examinations for comparison.
Results
- The results indicated that there was a 94% agreement rate between the two methods (PCR and immunohistochemistry), signifying both were almost equally effective in identifying EHV-1.
- When compared with the standard virology examination method, the PCR method’s agreement was 87%, and immunohistochemistry was 82%.
Conclusion
- The research found that both PCR and immunohistochemistry can effectively detect EHV-1 in paraffin-embedded tissues, making them powerful tools for this purpose.
- These methods provide results faster than traditional virus isolation methods.
- Moreover, they can detect inactivated (non-viable) virus, which are not identified by standard virus isolation methods.
- The use of these methods also allows for retrospective studies, presumably because they can be effective even on older, preserved samples.
- The researchers used two primary techniques to analyze the samples: a polymerase chain reaction (PCR) and immunohistochemistry.
- PCR allows for the amplification of specific DNA sequences for their detection. In this study, it was used to identify DNA fragments specific for EHV-1.
- Immunohistochemistry is a laboratory process used to visually detect antigens in tissue samples via the use of antibodies. In this case, it was used to identify EHV-1 antigens.
- 23 of the aborted fetus samples had also been tested with virology examinations for comparison.
Results
- The results indicated that there was a 94% agreement rate between the two methods (PCR and immunohistochemistry), signifying both were almost equally effective in identifying EHV-1.
- When compared with the standard virology examination method, the PCR method’s agreement was 87%, and immunohistochemistry was 82%.
Conclusion
- The research found that both PCR and immunohistochemistry can effectively detect EHV-1 in paraffin-embedded tissues, making them powerful tools for this purpose.
- These methods provide results faster than traditional virus isolation methods.
- Moreover, they can detect inactivated (non-viable) virus, which are not identified by standard virus isolation methods.
- The use of these methods also allows for retrospective studies, presumably because they can be effective even on older, preserved samples.
- The results indicated that there was a 94% agreement rate between the two methods (PCR and immunohistochemistry), signifying both were almost equally effective in identifying EHV-1.
- When compared with the standard virology examination method, the PCR method’s agreement was 87%, and immunohistochemistry was 82%.
Conclusion
- The research found that both PCR and immunohistochemistry can effectively detect EHV-1 in paraffin-embedded tissues, making them powerful tools for this purpose.
- These methods provide results faster than traditional virus isolation methods.
- Moreover, they can detect inactivated (non-viable) virus, which are not identified by standard virus isolation methods.
- The use of these methods also allows for retrospective studies, presumably because they can be effective even on older, preserved samples.
- The research found that both PCR and immunohistochemistry can effectively detect EHV-1 in paraffin-embedded tissues, making them powerful tools for this purpose.
- These methods provide results faster than traditional virus isolation methods.
- Moreover, they can detect inactivated (non-viable) virus, which are not identified by standard virus isolation methods.
- The use of these methods also allows for retrospective studies, presumably because they can be effective even on older, preserved samples.
Cite This Article
APA
Rimstad E, Evensen O.
(1993).
The identification of equid herpesvirus 1 in paraffin-embedded tissues from aborted fetuses by polymerase chain reaction and immunohistochemistry.
J Vet Diagn Invest, 5(2), 174-183.
https://doi.org/10.1177/104063879300500206 Publication
Researcher Affiliations
- Department of Microbiology and Immunology, Norwegian College of Veterinary Medicine, Oslo.
MeSH Terms
- Abortion, Veterinary / microbiology
- Animals
- Base Sequence
- DNA, Viral / analysis
- Electrophoresis, Polyacrylamide Gel / veterinary
- Female
- Fetus / microbiology
- Herpesviridae Infections / microbiology
- Herpesviridae Infections / pathology
- Herpesviridae Infections / veterinary
- Herpesvirus 1, Equid / isolation & purification
- Horse Diseases / microbiology
- Horse Diseases / pathology
- Horses
- Immunoblotting / veterinary
- Immunohistochemistry / methods
- Molecular Sequence Data
- Paraffin Embedding / veterinary
- Pilot Projects
- Polymerase Chain Reaction / methods
- Polymerase Chain Reaction / veterinary
- Pregnancy
- Sensitivity and Specificity
Citations
This article has been cited 4 times.- Taktaz Hafshejani T, Nekoei S, Vazirian B, Doosti A, Khamesipour F, Anyanwu MU. Molecular Detection of Equine Herpesvirus Types 1 and 4 Infection in Healthy Horses in Isfahan Central and Shahrekord Southwest Regions, Iran.. Biomed Res Int 2015;2015:917854.
- Molinková D, Celer V Jr, Jahn P. Isolation and partial characterization of equine herpesvirus type 1 in Czechia.. Folia Microbiol (Praha) 2004;49(5):605-11.
- Sutton GA, Viel L, Carman PS, Boag BL. Pathogenesis and clinical signs of equine herpesvirus-1 in experimentally infected ponies in vivo.. Can J Vet Res 1998 Jan;62(1):49-55.
- Pfeffer M, Wiedmann M, Batt CA. Applications of DNA amplification techniques in veterinary diagnostics.. Vet Res Commun 1995;19(5):375-407.
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