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.
Pomelova VG, Gaĭdamovich SIa, Demenev VA, Kadoshnikov IuP.A three-step concentration of Venezuelan equine encephalomyelitis (VEE) virus from tissue culture fluid was carried out in a two-phase system of polyethyleneglycol (PEG)--sodium dextran sulphate (SDS). The concentration method was based on the dependence of virus distribution coefficient upon NaCl content in the system which allowed alternating transfer of the virus from one phase of the system into the other. The infectious activity of the virus increased approximately 100-fold after the first step, 190-fold after the second, and 300-fold after the third step. The process of concentration was...
Skok MV, Denisiuk PV, Komissarenko SV.Glutaraldehyde treatment does not change the absorption of cytochrome c either in the visible or in UV spectra. It brings about the formation of dimers, trimers and high-polymeric forms of cytochrome c and shifts the pI of all cytochrome c isoelectric fractions to more acid pH. Polymerization also results in changes of kinetic parameters of cytochrome c benzidine reaction increasing its affinity to 3,3-diaminobenzidine with a simultaneous decrease in the effectiveness of H2O2 binding. These biochemical changes can be related to immunochemical differences of native and glutaraldehyde-treated cy...
The goal of this work was the development of suitable (real-time) RT-PCR techniques for fast and sensitive diagnosis of EAV and for molecular-epidemiological characterisation of viral strains, as an alternative to virus isolation. To this purpose two conventional RT-PCR methods and one real-time RT-PCR were adapted to detect the broadest possible spectrum of viral strains. Several dilutions with Bucyrus strain showed a 100-fold higher sensitivity of real-time RT-PCR and heminested RT-PCR compared to simple RT-PCR. Making use of 11 cell culture supernatants of different EAV isolates and 7 semen...
Ashby M, Moore R, King S, Newbrook K, Flannery J, Batten C.African horse sickness is a severe and often fatal disease affecting all species of equids. The aetiological agent, African horse sickness virus (AHSV), can be differentiated into nine serotypes. The identification of AHSV serotypes is vital for disease management, as this can influence vaccine selection and help trace disease incursion routes. In this study, we report the development and optimisation of a novel, molecular-based assay that utilises multiplex PCR and microsphere-based technology to expedite detection and differentiation of multiple AHSV serotypes in one assay. We demonstrated t...
Lobato OL, Nogueira TDS, Lima TET, Andrade FJDC, de Macedo MGG, Pereira RS, Xavier J, Amorim MR, Barbosa PP, da Rocha AS, Silva SDC, Alcantara LCJ....The cycle of the Orthoflavivirus nilense (West Nile virus - WNV) involves birds and mosquitoes, while humans and equids serve as terminal hosts. In 2014, the first human case in Brazil was confirmed in Piauí State. Objective: To investigate the presence of WNV in birds, mosquitoes, and equids in municipalities of Piauí. Methods: Collections were carried out following recommendations from the Ministry of Health of Brazil, in 11 municipalities (all with human cases or bird mortality), where biological samples were collected from birds, mosquitoes, and equids. The Viral RNA extraction was perfo...
Elsawy BSM, Alzan HF., a recently discovered tick-borne hemoparasite infecting equids globally, has significant implications for equine health. Although it is closely related to (sharing 23% genomic divergence), it establishes an asymptomatic carrier state in persistently infected horses, creating a silent transmission reservoir. Its discovery and unique genetics justify its classification as a new taxon. A critical diagnostic challenge is that the lack of the gene in prevents its detection by the standard cELISA, emphasizing the need for species-specific tools. Although species-specific PCR assays, including ...
Furukawa R, Tozaki T, Kawate K, Kikuchi M, Ishige T, Fukui E, Kakoi H.A subset of single-nucleotide polymorphisms (SNPs) are associated with various phenotypes, including diseases, coat colour, and athletic performance, and are widely used in medical and veterinary fields. In this study, we developed a novel method for direct SNP genotyping from whole blood without DNA extraction. Thoroughbred blood samples were diluted 100-fold with Milli-Q water and analysed using real-time polymerase chain reaction (PCR) with hydrolysis probes. Specificity and sensitivity were improved by increasing the annealing temperature and number of PCR cycles. Genotyping results for SN...
Kumar B, Brahmbhatt NN, Thakre B, Maharana BR, Parmar VL, Kumar M.Haemoparasitic diseases constitute a significant constraint to economic livestock farming. Diagnostic techniques that are inexpensive, rapid, reliable, and precise are crucial for the management of diseases. In this context, PCR assays are very valuable yet expensive since the samples must be processed before being included in the PCR reaction. Accordingly, the goal of the current study was to lower the PCR costs without jeopardizing the assay's sensitivity and specificity. For that purpose, the alkaline solution was optimized for low cost and quick DNA extraction (blood lysate), and PCR reage...
Songsungthong W, Pornthanakasem W, Leartsakulpanich U, Srijuntongsiri G.The ability to accurately and rapidly identify causative agents of infectious diseases facilitates precise treatment, improves clinical outcomes, and augments epidemiology studies. For many veterinary and zoonotic pathogens, however, simple molecular tests for species identification are not available. causes severe diseases, such as sleepy foal disease, septicemia, and meningitis in horses and pigs. can also cause severe diseases in humans bitten by infected animals. Existing identification methods are biochemical tests, 16S rRNA gene amplification followed by DNA sequencing, and MALDI-TOF ...
Langlands Z, Gubbins S, Carpenter S, England M.African horse sickness virus (AHSV: Sedoreoviridae; Orbivirus) causes a severe and often fatal disease in horses (African horse sickness: AHS) and is transmitted almost exclusively by Culicoides biting midges (Diptera: Ceratopogonidae). In recent years, unprecedented outbreaks of AHSV have occurred in new geographical foci in Thailand and other related Culicoides-borne viruses continue to emerge unexpectedly, causing disease outbreaks in northern Europe. This study investigated Culicoides abundance and diversity at a donkey (Equus asinus) sanctuary in southern England. The incidence and severi...
