Equine herpesvirus type 1: detection of viral DNA sequences in aborted fetuses with the polymerase chain reaction.
Abstract: Primers and probes were selected from the gene encoding glycoprotein 13 (gp 13) of equine herpesvirus 1 (EHV-1). The polymerase chain reaction (PCR) was run on infected and noninfected cultured cells and on 63 specimens from 29 aborted equine fetuses. The results were evaluated by electrophoresis and dot-blot hybridization using an oligonucleotide probe labeled with biotin. In the infected samples electrophoresis showed a PCR product of about 280 base pairs. The dot-blot hybridization confirmed that this product contained EHV-1 DNA sequences. PCR took 4 h and hybridization another 14 h; the results were thus achieved within 24 h and were highly specific for EHV-1. Close concordance was found between the results of PCR and virus isolation.
Publication Date: 1990-05-01 PubMed ID: 2163562DOI: 10.1016/0378-1135(90)90024-pGoogle Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research paper discusses the use of polymerase chain reaction (PCR) to detect equine herpesvirus type 1 (EHV-1) in aborted equine fetuses. The results indicate that the PCR method is quick, highly specific, and aligns well with traditional virus isolation methods.
Selection of Primers and Probes
- Primers and probes were selected from the gene encoding glycoprotein 13 (gp 13) of EHV-1.
- This is a strategic choice as gp 13 is an important genetic component of the EHV-1 virus.
Experimental Procedure
- The polymerase chain reaction (PCR) was used on infected and non-infected cultured cells as this technology allows for the amplification of DNA sequences.
- The PCR method was also performed on 63 specimens taken from 29 aborted equine fetuses for practical testing.
- Follwoing the PCR process, electrophoresis was employed. This technique separates DNA sequences based on their size and charge.
- The experiment also used dot-blot hybridization, a method that uses a labeled probe to detect specific DNA sequences.
- In this case, the utilized oligonucleotide probe was labeled with biotin for accurate detection.
Results and Conclusion
- The infected samples revealed a PCR product of about 280 base pairs during electrophoresis, while dot-blot hybridization confirmed that this product did contain EHV-1 DNA sequences.
- The entire PCR process and hybridization took about 18 hours, permitting results to be achieved within a single day.
- The results obtained from this quick PCR method were found to be highly specific for EHV-1 and presented close concordance with the standard method of virus isolation, attesting its validity as a detection method.
Cite This Article
APA
Ballagi-Pordány A, Klingeborn B, Flensburg J, Belák S.
(1990).
Equine herpesvirus type 1: detection of viral DNA sequences in aborted fetuses with the polymerase chain reaction.
Vet Microbiol, 22(4), 373-381.
https://doi.org/10.1016/0378-1135(90)90024-p Publication
Researcher Affiliations
- Department of Virology, National Veterinary Institute, Uppsala, Sweden.
MeSH Terms
- Abortion, Veterinary / diagnosis
- Animals
- DNA, Viral / analysis
- Electrophoresis, Agar Gel
- Female
- Fetus / microbiology
- Herpesviridae / genetics
- Herpesviridae Infections / diagnosis
- Herpesviridae Infections / veterinary
- Herpesvirus 1, Equid / genetics
- Herpesvirus 1, Equid / isolation & purification
- Horse Diseases / diagnosis
- Horses
- Nucleic Acid Hybridization
- Oligonucleotide Probes
- Placenta / microbiology
- Polymerase Chain Reaction
- Pregnancy
Citations
This article has been cited 11 times.- Oladunni FS, Horohov DW, Chambers TM. EHV-1: A Constant Threat to the Horse Industry. Front Microbiol 2019;10:2668.
- Smith KL, Li Y, Breheny P, Cook RF, Henney PJ, Sells S, Pronost S, Lu Z, Crossley BM, Timoney PJ, Balasuriya UB. New real-time PCR assay using allelic discrimination for detection and differentiation of equine herpesvirus-1 strains with A2254 and G2254 polymorphisms. J Clin Microbiol 2012 Jun;50(6):1981-8.
- Belák S. Molecular diagnosis of viral diseases, present trends and future aspects A view from the OIE Collaborating Centre for the Application of Polymerase Chain Reaction Methods for Diagnosis of Viral Diseases in Veterinary Medicine. Vaccine 2007 Jul 26;25(30):5444-52.
- Rodriguez JM. Detection of animal pathogens by using the polymerase chain reaction (PCR). Vet J 1997 May;153(3):287-305.
- Reubel GH, Ramos RA, Hickman MA, Rimstad E, Hoffmann DE, Pedersen NC. Detection of active and latent feline herpesvirus 1 infections using the polymerase chain reaction. Arch Virol 1993;132(3-4):409-20.
- Belák S, Ballagi-Pordány A. Application of the polymerase chain reaction (PCR) in veterinary diagnostic virology. Vet Res Commun 1993;17(1):55-72.
- Sinclair R, Binns MM, Chirnside ED, Mumford JA. Detection of antibodies against equine herpesvirus types 1 and 4 by using recombinant protein derived from an immunodominant region of glycoprotein B. J Clin Microbiol 1993 Feb;31(2):265-71.
- Yason CV, Harris LM, McKenna PK, Wadowska D, Kibenge FS. Establishment of conditions for the detection of bovine herpesvirus-1 by polymerase chain reaction using primers in the thymidine kinase region. Can J Vet Res 1995 Apr;59(2):94-101.
- Naeem K, Murtaugh MP, Goyal SM. Tissue distribution of bovid herpesvirus-4 in inoculated rabbits and its detection by DNA hybridization and polymerase chain reaction. Arch Virol 1991;119(3-4):239-55.
- Hohdatsu T, Yamada M, Okada M, Fukasawa M, Watanabe K, Ogasawara T, Takagi M, Aizawa C, Hayami M, Koyama H. Detection of feline immunodeficiency proviral DNA in peripheral blood lymphocytes by the polymerase chain reaction. Vet Microbiol 1992 Feb;30(2-3):113-23.
- Rimstad E, Hyllseth B. Equine herpesviruses 1 and 4: amplification and differentiation by polymerase chain reaction. Acta Vet Scand 1994;35(3):303-6.
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