Ashleigh C. McEvoy, Edith Cowan UniversityFollow
Leslie Calapre, Edith Cowan UniversityFollow
Michelle Pereira, Edith Cowan UniversityFollow
Tindaro M. Giardina
Muhammad A. Khattak
Tarek M. Meniawy
Antonia L. Pritchard
Nicholas K. Hayward
Michael J. Millward
Mel R. Ziman Dr, Edith Cowan UniversityFollow
Elin S. Gray, Edith Cowan UniversityFollow
Impact Journals LLC
Place of Publication
School of Medical and Health Sciences
National Health and Medical Research Council
Australian Government Research Training Program
Edith Cowan University “Inspiring Minds” scholarship
Cancer Research Trust
Western Australia Cancer Council grant (1100249)
Perpetual/Ramaciotti Health Investment grant
Department of Health WA
Spinnaker Health Research Foundation
NHMRC Numbers : 1046711, 1119791, 1117663
Currently mainly BRAF mutant circulating tumor DNA (ctDNA) is utilized to monitor patients with melanoma. TERT promoter mutations are common in various cancers and found in up to 70 % of melanomas, including half of BRAF wildtype cases. Therefore, a sensitive method for detection of TERT promoter mutations would increase the number of patients that could be monitored through ctDNA analysis.
A droplet digital PCR (ddPCR) assay was designed for the concurrent detection of chr5:1,295,228 C > T and chr5:1,295,250 C > T TERT promoter mutations. The assay was validated using 39 melanoma cell lines and 22 matched plasma and tumor samples. In addition, plasma samples from 56 metastatic melanoma patients and 56 healthy controls were tested for TERT promoter mutations.
The established ddPCR assay detected TERT promoter mutations with a lower limit of detection (LOD) of 0.17 %. Total concordance was demonstrated between ddPCR and Sanger sequencing in all cell lines except one, which carried a second mutation within the probe binding-site. Concordance between matched plasma and tumor tissue was 68 % (15/22), with a sensitivity of 53 % (95 % CI, 27 % - 79 %) and a specificity of 100 % (95 % CI, 59 % - 100 %). A significantly longer PFS (p = 0.028) was evident in ctDNA negative patients. Importantly, our TERT promoter mutations ddPCR assay allowed detection of ctDNA in 11 BRAF wild-type cases.
The TERT promoter mutation ddPCR assay offers a sensitive test for molecular analysis of melanoma tumors and ctDNA, with the potential to be applied to other cancers.
McEvoy, A. C. (2018). Circulating tumour DNA: A non-invasive biomarker for melanoma. Retrieved from http://ro.ecu.edu.au/theses/2064
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