title: Electrochemical Genetic Profiling of Single Cancer Cells.
abstract: 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 a simple, rapid, flexible, and inexpensive approach for the simultaneous quantification of multiple breast cancer related mRNA markers, with single tumor cell sensitivity.
authors.affiliation: Universitat Rovira i Virgili , Departament de Enginyeria Química, Av. Països Catalans 26, 43007 Tarragona, Spain.
authors.forename: Josep Ll
authors.initials: JL
authors.lastname: Acero Sánchez
authors.affiliation: Universitat Rovira i Virgili , Departament de Enginyeria Química, Av. Països Catalans 26, 43007 Tarragona, Spain.
authors.forename: Hamdi
authors.initials: H
authors.lastname: Joda
authors.affiliation: Universitat Rovira i Virgili , Departament de Enginyeria Química, Av. Països Catalans 26, 43007 Tarragona, Spain.
authors.forename: Olivier Y F
authors.initials: OY
authors.lastname: Henry
authors.affiliation: KTH Royal Institute of Technology , Science for Life Laboratory (SciLifeLab Stockholm), School of Biotechnology, Division of Gene Technology, SE-171 65 Solna, Sweden.
authors.forename: Beata W
authors.initials: BW
authors.lastname: Solnestam
authors.affiliation: KTH Royal Institute of Technology , Science for Life Laboratory (SciLifeLab Stockholm), School of Biotechnology, Division of Gene Technology, SE-171 65 Solna, Sweden.
authors.forename: Linda
authors.initials: L
authors.lastname: Kvastad
authors.affiliation: KTH Royal Institute of Technology , Science for Life Laboratory (SciLifeLab Stockholm), School of Biotechnology, Division of Gene Technology, SE-171 65 Solna, Sweden.
authors.forename: Pelin S
authors.initials: PS
authors.lastname: Akan
authors.affiliation: KTH Royal Institute of Technology , Science for Life Laboratory (SciLifeLab Stockholm), School of Biotechnology, Division of Gene Technology, SE-171 65 Solna, Sweden.
authors.forename: Joakim
authors.initials: J
authors.lastname: Lundeberg
authors.affiliation: MRC-Holland , Willem Schoutenstraat 1, 1057 DL Amsterdam, The Netherlands.
authors.forename: Nadja
authors.initials: N
authors.lastname: Laddach
authors.affiliation: Labman Automation Ltd. , Seamer Hill, Seamer, Stokesley, North Yorkshire TS9 5NQ, United Kingdom.
authors.forename: Dheeraj
authors.initials: D
authors.lastname: Ramakrishnan
authors.affiliation: Labman Automation Ltd. , Seamer Hill, Seamer, Stokesley, North Yorkshire TS9 5NQ, United Kingdom.
authors.forename: Ian
authors.initials: I
authors.lastname: Riley
authors.affiliation: Fraunhofer (ICT-IMM) , Carl-Zeiss-Str. 18-20, 55129 Mainz, Germany.
authors.forename: Carmen
authors.initials: C
authors.lastname: Schwind
authors.affiliation: Fraunhofer (ICT-IMM) , Carl-Zeiss-Str. 18-20, 55129 Mainz, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Latta
authors.affiliation: Universitat Rovira i Virgili , Departament de Enginyeria Química, Av. Països Catalans 26, 43007 Tarragona, Spain.|Institució Catalana de Recerca i Estudis Avançats , Passeig Lluís Companys 23, 08010 Barcelona, Spain.
authors.forename: Ciara K
authors.identifier: 0000-0003-2603-2230
authors.initials: CK
authors.lastname: O'Sullivan
mesh_terms: D015374:Biosensing Techniques; D001943:Breast Neoplasms / Q000235:genetics* / Q000473:pathology; D055664:Electrochemical Techniques; D005260:Female; D000076610:Genetic Profile; D006801:Humans; D009360:Neoplastic Cells, Circulating / Q000473:pathology*; D012333:RNA, Messenger / Q000235:genetics*; D059010:Single-Cell Analysis
chemical_list: D012333:RNA, Messenger