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Attomolar quantification of Mycobacterium tuberculosis by asymmetric helicase dependent isothermal DNA-amplification and electrochemical detection

dc.contributor.authorBarreda García, Susana 
dc.contributor.authorGonzález Álvarez, María José 
dc.contributor.authorSantos Álvarez, Noemí de los 
dc.contributor.authorPalacios Gutiérrez, Juan José 
dc.contributor.authorMiranda Ordieres, Arturo José 
dc.contributor.authorLobo Castañón, María Jesús 
dc.date.accessioned2015-08-20T07:27:45Z
dc.date.available2015-08-20T07:27:45Z
dc.date.issued2015
dc.identifier.citationBiosensors and Bioelectronics, 68, p. 122-128 (2014); doi:10.1016/j.bios.2014.12.029
dc.identifier.issn0956-5663
dc.identifier.issn1873-4235
dc.identifier.urihttp://hdl.handle.net/10651/32536
dc.description.abstractA highly sensitive and robust method for the quantification of specific DNA sequences based on coupling asymmetric helicase-dependent DNA amplification toelectrochemical detection is described.This method relies on the entrapment of the amplified ssDNA sequences on magnetic beads followed by a post-amplification hybridization assay to provide an added degree of specificity.Asaproof-of-concept a 84-bases long sequence specific of Mycobacterium tuberculosis is amplified at 65 °C, providing 3x10 6 amplification after 90 min.Using this system 0.5aM,corresponding to 15 copies of the target gene in 50 mL of sample,canbe successfully detected and reliably quantified under isothermal conditions in less than 4h.The assay has been applied to the detection of M. tuberculosis in sputum,pleural fluid andurine samples. Besides this application,the proposed assays is apowerful and general tool formoleculardi- agnostic that can be applied to the detection of other specific DNA sequences,taking full advantage of the plethora of genomic information now available.spa
dc.description.sponsorshipThis work has been supported by Ministerio de Economía y Competitividad (Spain)Projects CTQ2008-02429 and CTQ201231157,and the European Regional Development Fund.
dc.format.extentp. 122-128spa
dc.language.isoengspa
dc.publisherElsevier
dc.relation.ispartofBiosensors and Bioelectronics 68spa
dc.rights© 2014 Elsevier B.V.
dc.titleAttomolar quantification of Mycobacterium tuberculosis by asymmetric helicase dependent isothermal DNA-amplification and electrochemical detectionspa
dc.typejournal article
dc.identifier.doi10.1016/j.bios.2014.12.029
dc.relation.projectIDMINECO/CTQ2008-02429
dc.relation.projectIDMINECO/CTQ201231157
dc.relation.publisherversionhttp://dx.doi.org/10.1016/j.bios.2014.12.029


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