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Electrochemical Genetic Profiling of Single Cancer Cells
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2017 (Engelska)Ingår i: Analytical Chemistry, ISSN 0003-2700, E-ISSN 1520-6882, Vol. 89, nr 6, s. 3378-3385Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

Recent understandings in the development and spread of cancer have led to the realization of novel single cell analysis platforms focused on circulating tumor cells (CTCs). A simple, rapid, and inexpensive analytical platform capable of providing genetic information on these rare cells is highly desirable to support clinicians and researchers alike to either support the selection or adjustment of therapy or provide fundamental insights into cell function and cancer progression mechanisms. We report on the genetic profiling of single cancer cells, exploiting a combination of multiplex ligation-dependent probe amplification (MLPA) and electrochemical detection. Cells were isolated using laser capture and lysed, and the mRNA was extracted and transcribed into DNA. Seven markers were amplified by MLPA, which allows for the simultaneous amplification of multiple targets with a single primer pair, using MLPA probes containing unique barcode sequences. Capture probes complementary to each of these barcode sequences were immobilized on a printed circuit board (PCB) manufactured electrode array and exposed to single-stranded MLPA products and subsequently to a single stranded DNA reporter probe bearing a HRP molecule, followed by substrate addition and fast electrochemical pulse amperometric detection. We present asimple, rapid, flexible, and inexpensive approach for the simultaneous quantification of multiple breast cancer related mRNA markers, with single tumor cell sensitivity.

Ort, förlag, år, upplaga, sidor
American Chemical Society (ACS), 2017. Vol. 89, nr 6, s. 3378-3385
Nationell ämneskategori
Medicinsk bioteknologi (med inriktning mot cellbiologi (inklusive stamcellsbiologi), molekylärbiologi, mikrobiologi, biokemi eller biofarmaci)
Identifikatorer
URN: urn:nbn:se:kth:diva-205456DOI: 10.1021/acs.analchem.6b03973ISI: 000397478300021Scopus ID: 2-s2.0-85018371538OAI: oai:DiVA.org:kth-205456DiVA, id: diva2:1097271
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QC 20170522

Tillgänglig från: 2017-05-22 Skapad: 2017-05-22 Senast uppdaterad: 2017-05-22Bibliografiskt granskad

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Av författaren/redaktören
Kvastad, LindaSahlén, PelinLundeberg, Joakim
Av organisationen
GenteknologiScience for Life Laboratory, SciLifeLab
I samma tidskrift
Analytical Chemistry
Medicinsk bioteknologi (med inriktning mot cellbiologi (inklusive stamcellsbiologi), molekylärbiologi, mikrobiologi, biokemi eller biofarmaci)

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