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A new Reverse Transcription - Polymerase Chain Reaction (RT-PCR) assay is described that can differentiate between the nine serotypes of African horse sickness virus (AHSV). A unique set of primers targeting regions in the corresponding genome segment-2 were designed for each serotype. Sequences of the individual amplicons have single nucleotide polymorphisms (SNPs) allowing discrimination between field samples and either reference strains or live attenuated viruses (ALVs). This assay was tested and used to determine the serotype prevalence of AHSV samples submitted during the 2016 / 2017 season as well as to determine the incidence of field samples derived from ALVs. The information was used to study the epidemiology of AHSV in Southern Africa.
This article was published in the following journal.
Name: Journal of virological methods
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In vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short oligonucleotide flanking sequences (primers). The essential steps include thermal denaturation of the double-stranded target molecules, annealing of the primers to their complementary sequences, and extension of the annealed primers by enzymatic synthesis with DNA polymerase. The reaction is efficient, specific, and extremely sensitive. Uses for the reaction include disease diagnosis, detection of difficult-to-isolate pathogens, mutation analysis, genetic testing, DNA sequencing, and analyzing evolutionary relationships.
Methods for using more than one primer set in a polymerase chain reaction to amplify more than one segment of the target DNA sequence in a single reaction.
Methods used for detecting the amplified DNA products from the polymerase chain reaction as they accumulate instead of at the end of the reaction.
A heat stable DNA-DIRECTED DNA POLYMERASE from the bacteria Thermus aquaticus. It is widely used for the amplification of genes through the process of POLYMERASE CHAIN REACTION. EC 2.7.7.-.
A technique that labels specific sequences in whole chromosomes by in situ DNA chain elongation or PCR (polymerase chain reaction).
Polymerase Chain Reaction (PCR)
PCR (Polymerase Chain Reaction) uses the ability of DNA polymerase (enzymes that create DNA molecules by assembling nucleotides, the building blocks of DNA. These enzymes are essential to DNA replication and usually work in pairs to create two ident...
DNA sequencing is the process of determining the precise order of nucleotides within a DNA molecule. During DNA sequencing, the bases of a small fragment of DNA are sequentially identified from signals emitted as each fragment is re-synthesized from a ...