title: Detection of Streptococcus equi subspecies equi using a triplex qPCR assay.
abstract: Genome sequencing data for Streptococcus equi subspecies equi and zooepidemicus were used to develop a novel diagnostic triplex quantitative PCR (qPCR) assay targeting two genes specific to S. equi (eqbE and SEQ2190) and a unique 100 base pair control DNA sequence (SZIC) inserted into the SZO07770 pseudogene of S. zooepidemicus strain H70. This triplex strangles qPCR assay can provide results within 2h of sample receipt, has an overall sensitivity of 93.9% and specificity of 96.6% relative to the eqbE singlex assay and detects S. equi at levels below the threshold of the culture assay, even in the presence of contaminating bacteria.
authors.affiliation: Centre for Preventive Medicine, Animal Health Trust, Lanwades Park, Kentford, Newmarket, Suffolk CB8 7UU, UK.
authors.forename: Katy
authors.initials: K
authors.lastname: Webb
authors.forename: Colin
authors.initials: C
authors.lastname: Barker
authors.forename: Tihana
authors.initials: T
authors.lastname: Harrison
authors.forename: Zoe
authors.initials: Z
authors.lastname: Heather
authors.forename: Karen F
authors.initials: KF
authors.lastname: Steward
authors.forename: Carl
authors.initials: C
authors.lastname: Robinson
authors.forename: J Richard
authors.initials: JR
authors.lastname: Newton
authors.forename: Andrew S
authors.initials: AS
authors.lastname: Waller
mesh_terms: D004269:DNA, Bacterial / Q000235:genetics; D016680:Genome, Bacterial; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D018502:Streptococcus equi / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D004269:DNA, Bacterial