Circulating tumour cell RNA characterisation from colorectal cancer patient blood after inertial microfluidic enrichment
Winter M., Cai Z., Winkler K., Georgiou K., Inglis D., Lavranos T., Rezaei M., Ibragimi V. M. M., Thierry B.
MethodsX
Vol.6, P. 1512-1520
Опубликовано: 2019
Тип ресурса: Статья
DOI:10.1016/j.mex.2019.06.012
Аннотация:
The detection and molecular analysis of circulating tumour cells (CTCs) potentially provides a significant insight to the characterisation of disease, stage of progression and therapeutic options for cancer patients. Following on from the protocol by Warkiani et al. 2016, which describes a method of enriching CTCs from cancer patient blood with inertial microfluidics, we describe a method to measure the CTC RNA expression in the enriched fraction using droplet digital PCR and compare transcript detection with and without RNA pre-amplification. • Inertial microfluidics combined with droplet digital PCR is advantageous as it allows for CTC enrichment and subsequent RNA analysis from patient blood. This allows for patient tumour analysis with increased sensitivity and precision compared to quantitative Real Time PCR and enables the direct quantification of nucleic acids without the need for tumour biopsy. • This method demonstrates an efficient approach providing important insights into t
Ключевые слова:
Circulating tumour cell; Circulating tumour cells isolation and analysis with droplet digital PCR; Colorectal cancer; Droplet digital PCR
biological marker; complementary DNA; nucleic acid; RNA; Article; blood; blood sampling; cancer patient; cancer prognosis; circulating tumor cell; colorectal cancer; droplet digital polymerase chain reaction; gene expression assay; HT-29 cell line; human; human cell; LoVo cell line; microfluidics; priority journal; real time polymerase chain reaction; RNA extraction; tumor biopsy
Язык текста: Английский
ISSN: 2215-0161
Winter M.
Cai Z.
Winkler K.
Georgiou K.
Inglis D.
Lavranos T.
Rezaei M.
Ibragimi V. M. M. Varkiani Madzhid Modzhtaba 1983-
Thierry B.
Wинтер М.
Cаи З.
Wинклер К.
Георгиоу К.
Инглис Д.
Лавранос Т.
Резаеи М.
Ибрагими В. М. М. Варкиани Маджид Моджтаба 1983-
Тхиеррy Б.
Circulating tumour cell RNA characterisation from colorectal cancer patient blood after inertial microfluidic enrichment
Текст визуальный непосредственный
MethodsX
Vol.6 P. 1512-1520
2019
Статья
Circulating tumour cell Circulating tumour cells isolation and analysis with droplet digital PCR Colorectal cancer Droplet digital PCR
biological marker complementary DNA nucleic acid RNA Article blood blood sampling cancer patient cancer prognosis circulating tumor cell colorectal cancer droplet digital polymerase chain reaction gene expression assay HT-29 cell line human human cell LoVo cell line microfluidics priority journal real time polymerase chain reaction RNA extraction tumor biopsy
The detection and molecular analysis of circulating tumour cells (CTCs) potentially provides a significant insight to the characterisation of disease, stage of progression and therapeutic options for cancer patients. Following on from the protocol by Warkiani et al. 2016, which describes a method of enriching CTCs from cancer patient blood with inertial microfluidics, we describe a method to measure the CTC RNA expression in the enriched fraction using droplet digital PCR and compare transcript detection with and without RNA pre-amplification. • Inertial microfluidics combined with droplet digital PCR is advantageous as it allows for CTC enrichment and subsequent RNA analysis from patient blood. This allows for patient tumour analysis with increased sensitivity and precision compared to quantitative Real Time PCR and enables the direct quantification of nucleic acids without the need for tumour biopsy. • This method demonstrates an efficient approach providing important insights into t