Document Type

Journal Article


Impact Journals LLC

Place of Publication

United States


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

Grant Number

NHMRC Numbers : 1046711, 1119791, 1117663


McEvoy, A. C., Calapre, L., Pereira, M. R., Giardina, T., Robinson, C., Khattak, M. A., ... & Millward, M. (2017). Sensitive droplet digital PCR method for detection of TERT promoter mutations in cell free DNA from patients with metastatic melanoma. Oncotarget, 8, 78890-78900.



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.



Creative Commons License

Creative Commons Attribution 3.0 License
This work is licensed under a Creative Commons Attribution 3.0 License.

Included in

Oncology Commons