Analyze Diet

Topic:Polymerase Chain Reaction

Polymerase Chain Reaction (PCR) is a molecular biology technique used to amplify specific DNA sequences, allowing for detailed genetic analysis in horses. This method enables the detection and quantification of genetic material, facilitating research in areas such as genetic disorders, infectious diseases, and population genetics in equine species. PCR applications in horses include identifying pathogens, verifying parentage, and studying genetic variations. The technique's sensitivity and specificity make it a valuable tool in equine veterinary diagnostics and research. This page compiles peer-reviewed research studies and scholarly articles that explore the applications, methodologies, and advancements of PCR in equine science.
Epidemiological investigation of equid herpesvirus-4 (EHV-4) excretion assessed by nasal swabs taken from thoroughbred foals.
Veterinary microbiology    April 1, 1994   Volume 39, Issue 3-4 275-283 doi: 10.1016/0378-1135(94)90164-3
Gilkerson J, Jorm LR, Love DN, Lawrence GL, Whalley JM.Equid herpesvirus-4 (EHV-4) was detected in nasal swabs taken from foals using a PCR based test and this information used to study the epidemiology of EHV-4 disease on three Australian Thoroughbred stud farms in NSW in 1992. There was a very high level of agreement (kappa value of 0.84) between the PCR results and virus isolation using cell culture techniques. There was a strong seasonal distribution of EHV-4 shedding. Twenty-five of 26 positive samples were collected in January and March with the remaining positive sample collected in February. Foals with clinical signs of upper respiratory t...
In vivo replicative status and envelope heterogeneity of equine infectious anemia virus in an inapparent carrier.
Journal of virology    April 1, 1994   Volume 68, Issue 4 2777-2780 doi: 10.1128/JVI.68.4.2777-2780.1994
Kim CH, Casey JW.The distribution and replicative status of equine infectious anemia virus (EIAV) DNA in the tissues of a well-characterized inapparent carrier horse were established by using the PCR technique. The EIAV pol region could be amplified in all of the tissues tested, including the cerebellum and periventricular tissue, at concentrations approximately 10(5)-fold less than in the same tissue from an acutely infected horse. Further analysis of the EIAV genome, with primer pairs diagnostic for sequential stages of reverse transcription, suggests that EIAV DNA in the brain, liver, and lymph nodes was in...
Development and evaluation of PCR test for detection of Taylorella equigenitalis.
Journal of clinical microbiology    April 1, 1994   Volume 32, Issue 4 893-896 doi: 10.1128/jcm.32.4.893-896.1994
Bleumink-Pluym NM, Werdler ME, Houwers DJ, Parlevliet JM, Colenbrander B, van der Zeijst BA.A PCR for the detection of Taylorella equigenitalis, the causative agent of contagious equine metritis, was developed and evaluated. A genus-specific primer-probe set was derived from the 16S ribosomal DNA sequences. The PCR was specific and amplified a 585-bp product from all 64 available T. equigenitalis isolates. This PCR product hybridized with a specific probe in a dot spot assay. A variety of microorganisms from the genital tracts of horses or with a close phylogenetic relationship to T. equigenitalis did not yield a visible PCR product and were all negative in the dot spot hybridization...
Rapid, single-step differentiation of equid herpesviruses 1 and 4 from clinical material using the polymerase chain reaction and virus-specific primers.
Journal of virological methods    April 1, 1994   Volume 47, Issue 1-2 59-72 doi: 10.1016/0166-0934(94)90066-3
Lawrence GL, Gilkerson J, Love DN, Sabine M, Whalley JM.Sets of primers were designed which enabled specific amplification of homologous regions of the glycoprotein C and gene 76 genetic loci of equine herpesviruses 1 and 4 (EHV-1 and EHV-4). The resultant virus-specific polymerase chain reaction (PCR) products arising from each loci could be discriminated easily on the basis of size on an agarose gel, allowing rapid differentiation of the two equine herpesviruses. Specificity of the amplifications were confirmed by Southern hybridization and restriction endonuclease digestion. The PCR test was applied to nasal swab samples from weanling foals and ...
Alternative modes of polymerization distinguish the subunits of equine infectious anemia virus reverse transcriptase.
The Journal of biological chemistry    March 18, 1994   Volume 269, Issue 11 8541-8548 
Wöhrl BM, Howard KJ, Jacques PS, Le Grice SF.A comparative study of recombinant 51- and 66-kDa subunits comprising equine infectious anemia virus reverse transcriptase (EIAV RT) is reported. Both polypeptides sedimented as stable homodimers (molecular mass, 102 and 132 kDa, respectively) when analyzed by rate sedimentation through glycerol gradients. Consistent with their dimer composition, each preparation displayed considerable levels of both RNA- and DNA-dependent DNA polymerase activity on different homopolymeric template/primer combinations. However, a detailed analysis of the polymerization products indicated qualitative difference...
The prevalence of latent Equid herpesviruses in the tissues of 40 abattoir horses.
Equine veterinary journal    March 1, 1994   Volume 26, Issue 2 140-142 doi: 10.1111/j.2042-3306.1994.tb04353.x
Edington N, Welch HM, Griffiths L.Equid herpesviruses 1 or 4 (EHV-1 or -4) were isolated by cocultivation from 60% of 40 horses examined at slaughter. The lymph nodes draining the respiratory tract were the most common source of virus. EHV-1 or EHV-4 was never isolated from the trigeminal ganglia (SLG). The polymerase chain reaction (PCR) detected virus in 87.5% of bronchial lymph nodes and a similar level in the trigeminal ganglia that were examined. By both assays approximately one third of the positive animals harboured both viruses. Equid herpesvirus 2 (EHV-2) was isolated from all but one of the horses and from > 75% o...
Detection of equine arteritis virus following amplification of structural and nonstructural viral genes by reverse transcription-PCR.
Journal of clinical microbiology    March 1, 1994   Volume 32, Issue 3 658-665 doi: 10.1128/jcm.32.3.658-665.1994
St-Laurent G, Morin G, Archambault D.A reverse transcription (RT)-PCR assay was developed for the detection of equine arteritis virus (EAV) in cell culture supernatant and in horse semen. Four different sets of oligonucleotide primers complementary to sequences located in the 3' end of the polymerase gene (open reading frame [ORF] 1b) and to sequences representing the entire ORFs 3, 4, and 7, which encode for nonstructural (ORFs 3 and 4) or viral nucleocapsid (ORF 7) proteins, were compared for their abilities to amplify the targeted EAV sequences by the RT-PCR procedure. The sensitivities of the RT-PCR for amplification of EAV s...
Detection of African horse sickness virus by reverse transcription-PCR.
Journal of clinical microbiology    March 1, 1994   Volume 32, Issue 3 697-700 doi: 10.1128/jcm.32.3.697-700.1994
Stone-Marschat M, Carville A, Skowronek A, Laegreid WW.Reverse transcription-PCR (RT-PCR) was used to detect African horse sickness virus (AHSV). A single primer pair which amplified a 423-bp fragment of the S8 gene which encodes the NS2 protein of AHSV was identified. Amplification of this fragment from all nine serotypes of AHSV was achieved with these primers. Between 10(1) and 10(2) copies of AHSV genomic double-stranded RNA could be detected by RT-PCR followed by agarose gel electrophoresis and ethidium bromide staining. Application of RT-PCR to blood samples from AHSV-infected horses resulted in earlier detection of viremia than virus isolat...
Detection of Borna disease virus RNA in naturally infected animals by a nested polymerase chain reaction.
Journal of virological methods    February 1, 1994   Volume 46, Issue 2 133-143 doi: 10.1016/0166-0934(94)90098-1
Zimmermann W, Dürrwald R, Ludwig H.Borna disease virus in naturally infected horses, a donkey and sheep was detected for the first time by amplification of viral RNA using PCR. In contrast to a control group of healthy horses, brain tissue was positive by this assay in all animals with neurological symptoms. The use of a second round of PCR with nested primers following Southern hybridization confirmed the specificity and increased the sensitivity of the test. Comparison with conventional methods recommends this technique for monitoring of BDV infections at a molecular level.
The equine herpesvirus type 1 glycoprotein homologous to herpes simplex virus type 1 glycoprotein M is a major constituent of the virus particle.
The Journal of general virology    February 1, 1994   Volume 75 ( Pt 2) 439-442 doi: 10.1099/0022-1317-75-2-439
Pilling A, Davison AJ, Telford EA, Meredith DM.Glycoprotein 45 is a major envelope glycoprotein of equine herpesvirus type 1. The gene encoding this protein is located between map units 0.615 and 0.636 on the virus genome and evidence has suggested that it is encoded by gene 52, one of four genes within this region. Using PCR we have amplified gene 52 and subsequently cloned it into a mammalian expression vector under the control of the human cytomegalovirus immediate early gene promoter. The gene was expressed in COS-7 cells and its product was detected by immunofluorescence and Western blotting. The results indicate that glycoprotein 45 ...
Parentage testing and linkage analysis in the horse using a set of highly polymorphic microsatellites.
Animal genetics    February 1, 1994   Volume 25, Issue 1 19-23 
Marklund S, Ellegren H, Eriksson S, Sandberg K, Andersson L.Ten (TG)n positive clones, isolated from an equine genomic library and sequenced, contained 12-19 uninterrupted TG repeats. Primers for polymerase chain reaction (PCR) were synthesized and nine of these (TG)n loci (HTG7-15) were successfully amplified and utilized in this study together with five previously reported equine microsatellite loci (HTG2-6). The PCR products were analysed by polyacrylamide gel electrophoresis followed by automated laser fluorescence detection or autoradiography. All microsatellites showed polymorphism and stable Mendelian inheritance. Differences in microsatellite v...
Diagnosis of the African horse sickness virus serotype 4 by a one-tube, one manipulation RT-PCR reaction from infected organs.
Journal of virological methods    February 1, 1994   Volume 46, Issue 2 179-188 doi: 10.1016/0166-0934(94)90102-3
Zientara S, Sailleau C, Moulay S, Cruciere C.A single tube reverse transcription-polymerase chain reaction (RT-PCR) method for detection of African horse sickness virus (AHSV) in splenic tissues from infected horses is described. Double stranded RNA was extracted from infected organs of horses and used to produce complementary DNA (cDNA) with the two primers selected for the PCR. The 1179 bp amplified product (the segment 7 which encodes for VP 7), detected by electrophoresis on agarose gel and ethidium bromide staining, was hydrolysed with eight restriction endonucleases for characterization of the AHSV. The sensitivity of this method i...
Diagnosis of equine influenza by the polymerase chain reaction. Donofrio JC, Coonrod JD, Chambers TM.Influenza A is a common respiratory infection of horses, and rapid diagnosis is important for its detection and control. Sensitive detection of influenza currently requires viral culture and is not always feasible. The polymerase chain reaction (PCR) was used to detect DNA produced by reverse transcription of equine influenza in stored nasal secretions, vaccines, and allantoic fluids. Primers directed at a target of 212 bp on conserved segment 7 (matrix gene) of human influenza A/Bangkok/1/79(H3N2) produced amplification products of appropriate size with influenza A/Equine/Prague/1/56 (H7N7), ...
Molecular cloning and expression of equine interleukin 2.
Veterinary immunology and immunopathology    December 1, 1993   Volume 39, Issue 4 395-406 doi: 10.1016/0165-2427(93)90070-k
Vandergrifft EV, Horohov DW.We have cloned equine IL-2 cDNA in vitro using the polymerase chain reaction (PCR) and primers based on the human IL-2 sequence. The cloned product appears to contain the entire coding region for equine IL-2 based on homology with other known sequences. When expressed in COS cells, the recombinant product augmented the proliferative response of equine peripheral blood mononuclear cells to concanavalin A, however, it failed to support the continued proliferation of murine CTLL-2 cells. Specific substitutions in those regions associated with p55 and p75 binding appear to account for this species...
A rapid diagnostic assay for eastern equine encephalomyelitis viral RNA.
The American journal of tropical medicine and hygiene    December 1, 1993   Volume 49, Issue 6 772-776 doi: 10.4269/ajtmh.1993.49.772
Vodkin MH, McLaughlin GL, Day JF, Shope RE, Novak RJ.Eastern equine encephalomyelitis virus (EEEV) has been a low-frequency, but serious human and veterinary health problem. Increased frequency of this mosquito-borne virus is anticipated as wetlands are maintained and re-established. Control of EEEV has depended on mosquito abatement in response to increasing frequency of EEEV in the environment. A coupled reverse transcription/polymerase chain reaction assay was designed to rapidly, sensitively, and specifically detect EEEV RNA. The assay successfully detected the viral RNA in a single-blind study of a set of field samples composed of either po...
PCR-based detection of CEM agent.
The Veterinary record    October 9, 1993   Volume 133, Issue 15 375-376 doi: 10.1136/vr.133.15.375
Bleumink-Pluym NM, Houwers DJ, Parlevliet JM, Colenbrander B.No abstract available
Potomac horse fever.
The Veterinary clinics of North America. Equine practice    August 1, 1993   Volume 9, Issue 2 399-410 doi: 10.1016/s0749-0739(17)30406-6
Palmer JE.E. risticii, the cause of classic Potomac horse fever, is now known to produce two disease syndromes: EEC and EEA. The pathogen appears to commonly infect horses based on seroepidemiologic studies; however, the method of transmission remains unknown. The most common clinical disease is EEC, commonly called Potomac horse fever, which presents a wide spectrum of clinical signs. Diagnosis is currently dependent on serology, which frequently does not lead to a definitive diagnosis and at best results in a retrospective diagnosis. A new diagnostic approach, polymerase chain reaction, may offer a ra...
Apolipoprotein B mRNA editing in 12 different mammalian species: hepatic expression is reflected in low concentrations of apoB-containing plasma lipoproteins.
Journal of lipid research    August 1, 1993   Volume 34, Issue 8 1367-1383 
Greeve J, Altkemper I, Dieterich JH, Greten H, Windler E.Two different isoproteins are encoded by the apolipoprotein (apo) B gene, apoB-48 and apoB-100. ApoB-48, core component of intestinally derived chylomicrons, has an accelerated plasma turnover as compared with the full-length protein apoB-100. A posttranscriptional modification of the apoB mRNA by conversion of cytidine into uridine at nucleotide position 6666 changes the genomically encoded glutamine codon CAA at amino acid residue 2153 into a translational stop codon UAA. This mRNA editing explains the formation of the truncated isoform apoB-48. In the present investigation editing of apoB m...
Localization of xanthine dehydrogenase mRNA in horse skeletal muscle by in situ hybridization with digoxigenin-labelled probe.
The Biochemical journal    June 15, 1993   Volume 292 ( Pt 3), Issue Pt 3 639-641 doi: 10.1042/bj2920639
Räsänen LA, Karvonen U, Pösö AR.In situ hybridization was used to localize xanthine dehydrogenase (XDH) mRNA in horse skeletal muscle. Capillary endothelial cells were found to express XDH, but muscle cells did not give any signal. The digoxigenin-labelled probe was produced by PCR with primers based on the cDNA sequence of mouse XDH and horse lung cDNAs. A 4.3 kb mRNA was detected in a Northern blot.
DNA sequence analysis of serologically detected ELA class II haplotypes at the equine DQ beta locus.
Animal genetics    June 1, 1993   Volume 24, Issue 3 187-190 doi: 10.1111/j.1365-2052.1993.tb00285.x
Szalai G, Bailey E, Gerber H, Lazary S.The genetic diversity at the ELA DQ beta locus was investigated using polymerase chain reaction and DNA sequencing. Based upon serological methods 16 class II homozygous animals were selected and their genomic DNA was used. A DQ beta gene from an equine cDNA library was also sequenced. Our methodology and the similarity between the genomic and the cDNA sequences suggest that the studied locus is expressed on equine lymphocytes. In the predicted amino acid sequence the most extensive variation is located at residues 56-60. The pattern of these five amino acids is strongly correlated to the sero...
cDNA cloning of equine interleukin-2 by polymerase chain reaction.
Equine veterinary journal    May 1, 1993   Volume 25, Issue 3 242-243 doi: 10.1111/j.2042-3306.1993.tb02953.x
Tavernor AS, Allen WR, Butcher GW.No abstract available
The identification of equid herpesvirus 1 in paraffin-embedded tissues from aborted fetuses by polymerase chain reaction and immunohistochemistry. Rimstad E, Evensen O.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% (kap...
Analysis of multiple mRNAs from pathogenic equine infectious anemia virus (EIAV) in an acutely infected horse reveals a novel protein, Ttm, derived from the carboxy terminus of the EIAV transmembrane protein.
Journal of virology    February 1, 1993   Volume 67, Issue 2 832-842 doi: 10.1128/JVI.67.2.832-842.1993
Beisel CE, Edwards JF, Dunn LL, Rice NR.Transcription of pathogenic equine infectious anemia virus (EIAV) in an acutely infected horse was examined by using the polymerase chain reaction and nucleotide sequencing. Four spliced transcripts were identified in liver tissue, in contrast to the multiplicity of alternatively spliced messages reported for in vitro-propagated human immunodeficiency virus, simian immunodeficiency virus, and, to a lesser extent, EIAV. Nucleotide sequence analysis demonstrated that three of these mRNAs encode known viral proteins: the envelope precursor, the product of the S2 open reading frame, and the regula...
DNA of bovine papillomavirus type 1 and 2 in equine sarcoids: PCR detection and direct sequencing.
Archives of virology    January 1, 1993   Volume 132, Issue 1-2 121-131 doi: 10.1007/BF01309847
Otten N, von Tscharner C, Lazary S, Antczak DF, Gerber H.Nucleotide sequences of bovine papillomavirus (BPV) DNA amplified by the polymerase chain reaction (PCR) from samples of equine sarcoid skin tumours were determined. All naturally occurring sarcoids (n = 58 tumours from 32 horses and 2 donkeys) contained BPV-DNA. All but 3 of the genome fragments belonged to the BPV type 1 strain (BPV-1); the remaining were BPV type 2. Similar results were obtained with cutaneous bovine papillomas used as controls (n = 20). One of the horses, carrying 2 sarcoids, was particularly interesting; one tumour contained BPV-1 DNA whilst the other sarcoid yielded BPV-...
Genetic and antigenic analysis of an equine influenza H 3 isolate from the 1989 epidemic.
Archives of virology    January 1, 1993   Volume 130, Issue 1-2 33-43 doi: 10.1007/BF01318994
Binns MM, Daly JM, Chirnside ED, Mumford JA, Wood JM, Richards CM, Daniels RS.The haemagglutinin (HA) gene from the equine influenza H3N8 isolate Suffolk/89 has been cloned by reverse transcription and polymerase chain reaction amplification. The nucleotide sequence of the HA gene was determined from two independently cloned copies of the gene and was found to be most closely related to recent American isolates supporting the idea that most isolates of equine H3N8 are evolving as a single lineage. When the predicted amino acid sequence of the Suffolk/89 HA was examined, changes had taken place in at least four of the major antigenic sites, A, B, C, and D when compared t...
Abortion of virologically negative foetuses following experimental challenge of pregnant pony mares with equid herpesvirus 1.
Equine veterinary journal    July 1, 1992   Volume 24, Issue 4 256-259 doi: 10.1111/j.2042-3306.1992.tb02830.x
Smith KC, Whitwell KE, Binns MM, Dolby CA, Hannant D, Mumford JA.From 1988 to 1991, 51 pregnant pony mares were challenged intranasally or by aerosol with an isolate of EHV-1 (AB4) originally recovered from a quadriplegic mare. This resulted in 32 abortions, occurring from 9 to 29 days after infection. In 14 of the early abortions (Days 9-14), EHV-1 was not demonstrated in the foetal tissues by virus isolation or immunostaining despite no other non-viral cause for the abortion being evident. Application of the polymerase chain reaction to foetal tissues from 9 of these cases also proved negative. One of the 14 mares was destroyed immediately after abortion,...
Genomic variation and segregation of equine infectious anemia virus during acute infection.
Journal of virology    June 1, 1992   Volume 66, Issue 6 3879-3882 doi: 10.1128/JVI.66.6.3879-3882.1992
Kim CH, Casey JW.Equine infectious anemia virus (EIAV) is a lentivirus that infects and persists in the monocyte/macrophage populations of blood and tissues. We employed polymerase chain reaction to investigate the distribution and the level of genome variability of EIAV DNA in different tissues of a horse infected with a highly virulent variant of the Wyoming strain of the virus. Long terminal repeat, gag, and pol primer pairs were used to direct the amplification of EIAV DNA from the peripheral blood mononuclear cells and from cells, presumably the macrophage subtypes, of the kidney, spleen, liver, lymph nod...
A single gene encodes the beta-subunits of equine luteinizing hormone and chorionic gonadotropin.
Molecular endocrinology (Baltimore, Md.)    June 1, 1992   Volume 6, Issue 6 951-959 doi: 10.1210/mend.6.6.1379674
Sherman GB, Wolfe MW, Farmerie TA, Clay CM, Threadgill DS, Sharp DC, Nilson JH.Equine (e) CG and LH beta-subunits have identical amino acid sequences, including an extended carboxyl-terminal peptide (CTP). This suggests that unlike the corresponding human genes, the beta-subunits of eCG and eLH may be encoded by a single gene and share a common proximal promotor region. To explore this, we isolated and characterized the eLH/CG beta gene(s). Data from Southern analyses suggest that the eCG beta and eLH beta subunits are products of the same single copy gene (eLH/CG beta). Overlapping fragments of the eLH/CG beta gene and cDNA were amplified from equine genomic DNA and pit...
Latent equid herpesviruses 1 and 4: detection and distinction using the polymerase chain reaction and co-cultivation from lymphoid tissues.
The Journal of general virology    February 1, 1992   Volume 73 ( Pt 2) 261-268 doi: 10.1099/0022-1317-73-2-261
Welch HM, Bridges CG, Lyon AM, Griffiths L, Edington N.The polymerase chain reaction (PCR) and co-cultivation were used to identify the lymphoreticular system as the site of latency of equid herpesvirus I (EHV-1). Primers for PCR were designed from aligned nucleotide sequences of the glycoprotein gB genes to amplify the same region of both the EHV-1 and EHV-4 genomes. Subsequent restriction digests using specific enzymes distinguished the amplified fragments of the EHV-1 genome from those of the EHV-4 genome. Ten weeks following an experimental infection of five ponies with EHV-1, latent virus was detected by PCR and recovered by co-cultivation, p...
[The polymerase chain reaction (PCR) for the detection of DNA of equine herpesviruses 1 and 4].
Berliner und Munchener tierarztliche Wochenschrift    February 1, 1992   Volume 105, Issue 2 52-55 
Hardt M, Teifke JP, Weiss E.Formalin-fixed and Paraplast-embedded tissue samples of 42 aborted equine fetuses were examined by polymerase chain reaction for the presence of equine herpesvirus DNA. The used set of primers was located in the glycoprotein 13 open reading frame and allowed the amplification of both EHV 1 und EHV 4. By cleaving pattern analysis after Hinf I digestion EHV 1 could be distinguished from EHV 4. In 9 of the cases investigated EHV 1-DNA was detected. This finding is in absolute context with the results of the virological investigations.