Sensitive droplet digital PCR method for detection of TERT promoter mutations in cell free DNA from patients with metastatic melanoma

被引：41

作者：

McEvoy, Ashleigh C. ^{[1
]}

Calapre, Leslie ^{[1
]}

Pereira, Michelle R. ^{[1
]}

Giardina, Tindaro ^{[2
]}

Robinson, Cleo ^{[2
,3
,4
]}

Khattak, Muhammad A. ^{[3
,5
]}

Meniawy, Tarek M. ^{[3
,6
]}

Pritchard, Antonia L. ^{[7
]}

Hayward, Nicholas K. ^{[7
]}

Amanuel, Benhur ^{[2
,3
]}

Millward, Michael ^{[3
,6
]}

Ziman, Melanie ^{[1
,4
]}

Gray, Elin S. ^{[1
]}

机构：

[1] Edith Cowan Univ, Sch Med & Hlth Sci, Joondalup, WA, Australia

[2] QEII Med Ctr, Anat Pathol, PathWest, Nedlands, WA, Australia

[3] Univ Western Australia, Sch Med & Pharmacol, Crawley, WA, Australia

[4] Univ Western Australia, Sch Pathol & Lab Med, Crawley, WA, Australia

[5] Fiona Stanley Hosp, Dept Med Oncol, Murdoch, WA, Australia

[6] Sir Charles Gairdner Hosp, Dept Med Oncol, Nedlands, WA, Australia

[7] QIMR Berghofer Med Res Inst, Brisbane, Qld, Australia

来源：

ONCOTARGET | 2017年 / 8卷 / 45期

基金：

澳大利亚国家健康与医学研究理事会; 英国医学研究理事会;

关键词：

droplet digital PCR (ddPCR); TERT; melanoma; cancer; CIRCULATING TUMOR DNA; COLORECTAL-CANCER; KRAS MUTATIONS; ACQUIRED-RESISTANCE; BREAST-CANCER; BRAF; THERAPY; GLIOBLASTOMA; PROGRESSION; XENOGRAFTS;

D O I：

10.18632/oncotarget.20354

中图分类号：

R73 [肿瘤学];

学科分类号：

100214 ;

摘要：

Background: 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. Methods: 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. Results: 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. Conclusions: 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.

引用

页码：78890 / 78900

页数：11

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