BibTex format
@article{Kalofonou:2020:10.1038/s41598-020-60852-3,
author = {Kalofonou, M and Malpartida-Cardenas, K and Alexandrou, G and Rodriguez-Manzano, J and Yu, L-S and Miscourides, N and Allsopp, R and LT, Gleason K and Goddard, K and Fernandez-Garcia, D and Page, K and Georgiou, P and Ali, S and Coombes, RC and Shaw, J and Toumazou, C},
doi = {10.1038/s41598-020-60852-3},
journal = {Scientific Reports},
title = {A novel hotspot specific isothermal amplification method for detection of thecommon PIK3CA p.H1047R breast cancer mutation},
url = {http://dx.doi.org/10.1038/s41598-020-60852-3},
volume = {10},
year = {2020}
}
RIS format (EndNote, RefMan)
TY - JOUR
AB - Breast cancer (BC) is a common cancer in women worldwide. Despite advances in treatment, up to 30% of women eventually relapse and die of metastatic breast cancer. Liquid biopsy analysis of circulating cell-free DNA fragments in the patients’ blood can monitor clonality and evolving mutations as a surrogate for tumour biopsy. Next generation sequencing platforms and digital droplet PCR can be used to profile circulating tumour DNA from liquid biopsies; however, they are expensive and time consuming for clinical use. Here, we report a novel strategy with proof-of-concept data that supports the usage of loop-mediated isothermal amplification (LAMP) to detect PIK3CA c.3140 A > G (H1047R), a prevalent BC missense mutation that is attributed to BC tumour growth. Allele-specific primers were designed and optimized to detect the p.H1047R variant following the USS-sbLAMP method. The assay was developed with synthetic DNA templates and validated with DNA from two breast cancer cell-lines and two patient tumour tissue samples through a qPCR instrument and finally piloted on an ISFET enabled microchip. This work sets a foundation for BC mutational profiling on a Lab-on-Chip device, to help the early detection of patient relapse and to monitor efficacy of systemic therapies for personalised cancer patient management.
AU - Kalofonou,M
AU - Malpartida-Cardenas,K
AU - Alexandrou,G
AU - Rodriguez-Manzano,J
AU - Yu,L-S
AU - Miscourides,N
AU - Allsopp,R
AU - LT,Gleason K
AU - Goddard,K
AU - Fernandez-Garcia,D
AU - Page,K
AU - Georgiou,P
AU - Ali,S
AU - Coombes,RC
AU - Shaw,J
AU - Toumazou,C
DO - 10.1038/s41598-020-60852-3
PY - 2020///
SN - 2045-2322
TI - A novel hotspot specific isothermal amplification method for detection of thecommon PIK3CA p.H1047R breast cancer mutation
T2 - Scientific Reports
UR - http://dx.doi.org/10.1038/s41598-020-60852-3
UR - http://hdl.handle.net/10044/1/77961
VL - 10
ER -