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dc.contributor.authorMbano, Ian M.
dc.contributor.authorMandizvo, Tawanda
dc.contributor.authorRogich, Jerome
dc.contributor.authorKunota, Tafara T. R.
dc.contributor.authorMackenzie, Jared S.
dc.contributor.authorPillay, Manormoney
dc.contributor.authorBalagadde, Frederick K.
dc.date2022-08-11T08:09:56.000
dc.date.accessioned2022-08-23T16:49:28Z
dc.date.available2022-08-23T16:49:28Z
dc.date.issued2020-05-01
dc.date.submitted2020-06-09
dc.identifier.citation<p>Mbano IM, Mandizvo T, Rogich J, Kunota TTR, Mackenzie JS, Pillay M, Balagaddé FK. Light Forge: A Microfluidic DNA Melting-based Tuberculosis Test. J Appl Lab Med. 2020 May 1;5(3):440-453. doi: 10.1093/jalm/jfaa019. PMID: 32445364; PMCID: PMC7192548. <a href="https://doi.org/10.1093/jalm/jfaa019">Link to article on publisher's site</a></p>
dc.identifier.issn2475-7241 (Linking)
dc.identifier.doi10.1093/jalm/jfaa019
dc.identifier.pmid32445364
dc.identifier.urihttp://hdl.handle.net/20.500.14038/41468
dc.description.abstractBACKGROUND: There is a well-documented lack of rapid, low-cost tuberculosis (TB) drug resistance diagnostics in low-income settings across the globe. It is these areas that are plagued with a disproportionately high disease burden and in greatest need of these diagnostics. METHODS: In this study, we compared the performance of Light Forge, a microfluidic high-resolution melting analysis (HRMA) prototype for rapid low-cost detection of TB drug resistance with a commercial HRMA device, a predictive "nearest-neighbor" thermodynamic model, DNA sequencing, and phenotypic drug susceptibility testing (DST). The initial development and assessment of the Light Forge assay was performed with 7 phenotypically drug resistant strains of Mycobacterium tuberculosis (M.tb) that had their rpoB gene subsequently sequenced to confirm resistance to Rifampin. These isolates of M.tb were then compared against a drug-susceptible standard, H37Rv. Seven strains of M.tb were isolated from clinical specimens and individually analyzed to characterize the unique melting profile of each strain. RESULTS: Light Forge was able to detect drug-resistance linked mutations with 100% concordance to the sequencing, phenotypic DST and the "nearest neighbor" thermodynamic model. Researchers were then blinded to the resistance profile of the seven M.tb strains. In this experiment, Light Forge correctly classified 7 out of 9 strains as either drug resistant or drug susceptible. CONCLUSIONS: Light Forge represents a promising prototype for a fast, low-cost diagnostic alternative for detection of drug resistant strains of TB in resource constrained settings.
dc.language.isoen_US
dc.relation<p><a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&list_uids=32445364&dopt=Abstract">Link to Article in PubMed</a></p>
dc.rightsCopyright American Association for Clinical Chemistry 2020. This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs licence (http://creative commons.org/licenses/by-nc-nd/4.0/), which permits non-commercial reproduction and distribution of the work, in any medium, provided the original work is not altered or transformed in any way, and that the work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/
dc.subjectMycobacterium tuberculosis
dc.subjectDrug resistance
dc.subjectHigh-resolution melting analysis
dc.subjectMolecular diagnostics
dc.subjectReal-time PCR
dc.subjectBacterial Infections and Mycoses
dc.subjectDiagnosis
dc.subjectHealth Services Administration
dc.subjectHealth Services Research
dc.subjectInternational Public Health
dc.subjectMicrobiology
dc.titleLight Forge: A Microfluidic DNA Melting-based Tuberculosis Test
dc.typeJournal Article
dc.source.journaltitleThe journal of applied laboratory medicine
dc.source.volume5
dc.source.issue3
dc.identifier.legacyfulltexthttps://escholarship.umassmed.edu/cgi/viewcontent.cgi?article=5265&amp;context=oapubs&amp;unstamped=1
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/oapubs/4246
dc.identifier.contextkey18035208
refterms.dateFOA2022-08-23T16:49:28Z
html.description.abstract<p>BACKGROUND: There is a well-documented lack of rapid, low-cost tuberculosis (TB) drug resistance diagnostics in low-income settings across the globe. It is these areas that are plagued with a disproportionately high disease burden and in greatest need of these diagnostics.</p> <p>METHODS: In this study, we compared the performance of Light Forge, a microfluidic high-resolution melting analysis (HRMA) prototype for rapid low-cost detection of TB drug resistance with a commercial HRMA device, a predictive "nearest-neighbor" thermodynamic model, DNA sequencing, and phenotypic drug susceptibility testing (DST). The initial development and assessment of the Light Forge assay was performed with 7 phenotypically drug resistant strains of Mycobacterium tuberculosis (M.tb) that had their rpoB gene subsequently sequenced to confirm resistance to Rifampin. These isolates of M.tb were then compared against a drug-susceptible standard, H37Rv. Seven strains of M.tb were isolated from clinical specimens and individually analyzed to characterize the unique melting profile of each strain.</p> <p>RESULTS: Light Forge was able to detect drug-resistance linked mutations with 100% concordance to the sequencing, phenotypic DST and the "nearest neighbor" thermodynamic model. Researchers were then blinded to the resistance profile of the seven M.tb strains. In this experiment, Light Forge correctly classified 7 out of 9 strains as either drug resistant or drug susceptible.</p> <p>CONCLUSIONS: Light Forge represents a promising prototype for a fast, low-cost diagnostic alternative for detection of drug resistant strains of TB in resource constrained settings.</p>
dc.identifier.submissionpathoapubs/4246
dc.contributor.departmentSchool of Medicine
dc.source.pages440-453


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Copyright American Association for Clinical Chemistry 2020. This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs licence (http://creative commons.org/licenses/by-nc-nd/4.0/), which permits non-commercial reproduction and distribution of the work, in any medium, provided the original work is not altered or transformed in any way, and that the work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
Except where otherwise noted, this item's license is described as Copyright American Association for Clinical Chemistry 2020. This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs licence (http://creative commons.org/licenses/by-nc-nd/4.0/), which permits non-commercial reproduction and distribution of the work, in any medium, provided the original work is not altered or transformed in any way, and that the work is properly cited. For commercial re-use, please contact journals.permissions@oup.com