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dc.contributor.authorXaio, Han
dc.contributor.authorGillespie, Stephen H
dc.contributor.editorGillespie, Stephen
dc.identifier.citationXaio , H & Gillespie , S H 2018 , Using RT qPCR for quantifying Mycobacteria marinum from in vitro and in vivo samples . in S Gillespie (ed.) , Antibiotic Resistance Protocols . Methods in Molecular Biology , vol. 1736 , Humana Press/Springer , New York, NY , pp. 137-145 .
dc.identifier.otherPURE: 252482706
dc.identifier.otherPURE UUID: f8554d00-af41-4ac1-b3cd-e3bb7b99d43d
dc.identifier.otherPubMed: 29322466
dc.identifier.otherScopus: 85040695699
dc.identifier.otherORCID: /0000-0001-6537-7712/work/42023882
dc.description.abstractMycobacterium marinum, the causative agent of fish tuberculosis, is rarely a human pathogen causing a chronic skin infection. It is now wildely used as a model system in animal models, especially in zebra fish model, to study the pathology of tuberculosis and as a means of screening new anti-tuberculosis agent. To facilitate such research, quantifying the viable count of M. marinum bacteria is a crucial step. The main approach used currently is still by counting the number of colony forming units (cfu), a method that has been in place for almost 100 years. Though this method well established, understood and relatively easy to perform, it is time-consuming and labor-intensive. The result can be compromised by failure to grow effectively and the relationship between count and actual numbers is confused by clumping of the bacteria where a single colony is made from multiple organisms. More importantly, this method is not able to detect live but not cultivable bacteria, and there is increasing evidence that mycobacteria readily enter a "dormant" state which confounds the relationship between bacterial number in the host and the number detected in a cfu assay. DNA based PCR methods detect both living and dead organisms but here we describe a method, which utilizes species specific Taq-Man assay and RT-qPCR technology for quantifying the viable M. marinum bacterial load by detecting 16S ribosomal RNA (16S rRNA).en
dc.publisherHumana Press/Springer
dc.relation.ispartofAntibiotic Resistance Protocolsen
dc.relation.ispartofseriesMethods in Molecular Biologyen
dc.rights© 2018, Springer Science+Business Media, LLC. This work has been made available online by kind permission of the publisher. This is the author created, accepted version manuscript following peer review and may differ slightly from the final published version. The final published version of this work is available at
dc.subjectQH301 Biologyen
dc.subjectRM Therapeutics. Pharmacologyen
dc.titleUsing RT qPCR for quantifying Mycobacteria marinum from in vitro and in vivo samplesen
dc.typeBook itemen
dc.contributor.institutionUniversity of St Andrews. School of Medicineen
dc.contributor.institutionUniversity of St Andrews. Infection and Global Health Divisionen
dc.contributor.institutionUniversity of St Andrews. Global Health Implementation Groupen
dc.contributor.institutionUniversity of St Andrews. Gillespie Groupen
dc.contributor.institutionUniversity of St Andrews. Biomedical Sciences Research Complexen
dc.contributor.institutionUniversity of St Andrews. Infection Groupen

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