Electrochemical Genetic Profiling of Single Cancer Cells

Autor: Ian Riley, Linda Kvastad, Carmen Schwind, Daniel Latta, Joakim Lundeberg, Nadja Laddach, Ciara K. O'Sullivan, Josep Ll. Acero Sánchez, Beata Werne Solnestam, Pelin Akan, Olivier Y.F. Henry, Hamdi Joda, Dheeraj Ramakrishnan
Přispěvatelé: Publica
Rok vydání: 2017
Předmět:
0301 basic medicine
DNA sequences
single-stranded DNA
Biosensing Techniques
Barcode
01 natural sciences
Analytical Chemistry
law.invention
Circulating tumor cell
Single-cell analysis
law
Multiplex
Chemistry
Printed Circuit Boards (PCB)
Genetic Profile
Neoplastic Cells
Circulating
3. Good health
DNA profiling
electrochemical detection
Female
Single-Cell Analysis
single cell analysis
Breast Neoplasms
Computational biology
probe
circulating tumor cells
010402 general chemistry
pulse amperometric detections
diseases
03 medical and health sciences
medicine
Humans
Multiplex ligation-dependent probe amplification
RNA
Messenger
gene
substrates
Tumors
Bar codes
genetic information
chemical detection
Cancer
DNA
Electrochemical Techniques
medicine.disease
probe amplification
Molecular biology
0104 chemical sciences
nucleic acid
030104 developmental biology
Cancer cell
Zdroj: Analytical chemistry. 89(6)
ISSN: 1520-6882
Popis: 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.
Databáze: OpenAIRE
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