title: Development of a loop-mediated isothermal amplification method for detecting Streptococcus equi subsp. zooepidemicus and analysis of its use with three simple methods of extracting DNA from equine respiratory tract specimens.
abstract: Streptococcus equi subsp. zooepidemicus (S. zooepidemicus) is a dominant pathogenic bacterium in equine pneumonia. We developed a specific loop-mediated isothermal amplification (LAMP) method, which targets the gene encoding sorbitol-6-phosphate 2-dehydrogenase (sorD), for detecting S. zooepidemicus and examined the clinical efficacies of its use in combination with each of 3 DNA extraction methods easily used by veterinary practitioners, namely the Loopamp PURE DNA Extraction Kit, InstaGene Matrix and a conventional boiling method. The LAMP method plus the Loopamp PURE DNA Extraction Kit gave higher rates of positivity than the other combinations in both clinical and spiked samples containing clinically significant concentrations (>1 × 10(4) CFU/ml) of S. zooepidemicus.
authors.affiliation: Microbiology Division, Epizootic Research Center, Equine Research Institute, Japan Racing Association, 1400-4 Shiba, Shimotuke, Tochigi 329-0412, Japan.
authors.forename: Yuta
authors.initials: Y
authors.lastname: Kinoshita
authors.forename: Hidekazu
authors.initials: H
authors.lastname: Niwa
authors.forename: Yoshinari
authors.initials: Y
authors.lastname: Katayama
mesh_terms: D000818:Animals; D015169:Colony Count, Microbial / Q000662:veterinary; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D006734:Horse Diseases / Q000175:diagnosis / Q000382:microbiology*; D006736:Horses; D057230:Limit of Detection; D021141:Nucleic Acid Amplification Techniques / Q000379:methods / Q000662:veterinary*; D011014:Pneumonia / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012680:Sensitivity and Specificity; D013290:Streptococcal Infections / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D018502:Streptococcus equi / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D004269:DNA, Bacterial