About: Characterization of a novel Mycoplasma cynos real-time PCR assay

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An Entity of Type : schema:ScholarlyArticle, within Data Space : covidontheweb.inria.fr associated with source document(s)

Attributes	Values
type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
has title	Characterization of a novel Mycoplasma cynos real-time PCR assay
Creator	Thachil, Anil Goodman, Laura Anderson, Renee Glaser, Amy Forbes, Zachary Gioia, Gloria Mitchell, Patrick Moroni, Paolo Tallmadge, Rebecca Werner, Brenda
Source	Medline; PMC
abstract	Mycoplasma cynos is recognized as an emerging causative pathogen of canine infectious respiratory disease (CIRD) worldwide. We developed a new open-source real-time PCR (rtPCR) assay for M. cynos that performs well under standard rtPCR conditions. Primers and probes were designed to target the M. cynos tuf gene. Reaction efficiencies for the M. cynos tuf gene assay on 2 platforms were based on amplification of standard curves spanning 8 orders of magnitude: ABI 7500 platform, 94.3–97.9% (r(2) ≥ 0.9935); QuantStudio OpenArray platform, 119.1–122.5% (r(2) = 0.9784). The assay performed very well over a range of template input, from 10(9) copies to the lower limit of quantification at 4 copies of the M. cynos genome on the ABI 7500 platform. Diagnostic performance was estimated by comparison with an in-house legacy assay on clinical specimens as well as testing isolates that were characterized previously by intergenic spacer region (ISR) sequencing. Exclusivity was established by testing 12 other Mycoplasma species. To substantiate the high specificity of the M. cynos tuf gene assay, sequence confirmation was performed on ISR PCR amplicons obtained from clinical specimens. One ISR amplicon sequence revealed M. mucosicanis rather than M. cynos. The complete protocol of the newly developed M. cynos tuf assay is provided to facilitate assay harmonization.
has issue date	2019-11-21(xsd:dateTime)
bibo:doi	10.1177/1040638719890858
bibo:pmid	31752630
has license	cc-by
sha1sum (hex)	ace6d9fe4d7fc6adfaa1aac8593466e0ace5b4e3
schema:url	https://doi.org/10.1177/1040638719890858
resource representing a document's title	Characterization of a novel Mycoplasma cynos real-time PCR assay
has PubMed Central identifier	PMC7206559
has PubMed identifier	31752630
schema:publication	J Vet Diagn Invest
resource representing a document's body	covid:ace6d9fe4d7fc6adfaa1aac8593466e0ace5b4e3#body_text
is schema:about of	named entity 'developed' named entity 'assay' named entity 'probes' named entity 'Mycoplasma' named entity 'COPIES' named entity 'RANGE' named entity 'CONFIRMATION' named entity 'NEWLY' named entity 'TARGET' named entity 'INPUT' named entity 'MAGNITUDE' named entity 'RATHER' named entity 'AMPLIFICATION' named entity 'testing' named entity 'ABI' named entity 'spanning' named entity 'legacy' named entity 'high' named entity 'species' named entity 'sequence' named entity 'revealed' named entity 'pathogen' named entity 'assay' named entity 'canine' named entity 'cynos' named entity 'genome' named entity 'amplicon' named entity 'PCR' named entity 'Mycoplasma' named entity 'gene' named entity 'rtPCR' named entity 'rtPCR' named entity 'amplicons' named entity 'orders of magnitude' named entity 'cynos' named entity 'assay' named entity 'Mycoplasma' named entity 'assay' named entity 'University of Milan' named entity 'canine' named entity 'pneumonia' named entity 'Integrated DNA Technologies' named entity 'Mycoplasma' named entity 'M. canis' named entity 'Pathogens' named entity 'cynos' named entity 'viruses' named entity 'canine' named entity 'ISR' named entity 'phylogenetic' named entity 'M. spumans' named entity 'ISR' named entity 'Thermo Fisher Scientific' named entity 'Redmond, WA' named entity 'PCR' named entity 'canine' named entity 'Mycoplasma' named entity 'high-throughput' named entity 'fluorometer' named entity 'equine' named entity 'cynos' named entity 'genome' named entity 'coinfections' named entity 'coinfection' named entity 'neighbor-joining method' named entity 'limit of quantification' named entity '10.7' named entity 'canine' named entity 'respiratory disease' named entity 'nasal swab' named entity 'genome sequences' named entity 'Mycoplasma'

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