Quantitative analysis of the BRAF
                     V600E mutation in circulating tumor-derived DNA in melanoma patients using competitive allele-specific TaqMan PCR

Ashida, Atsuko; Sakaizawa, Kaori; Mikoshiba, Asuka; Uhara, Hisashi; Okuyama, Ryuhei

doi:10.1007/s10147-016-0976-y

Cited by 35 publications

(28 citation statements)

References 25 publications

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“…Based on PCR technology, new technologies including real‐time quantitative PCR (qPCR) (Brown, ), amplification‐refractory mutation system (ARMS)‐based qPCR (Zhang et al ., ), competitive allele‐specific TaqMan PCR (cast‐PCR) (Ashida et al ., ; Reid et al ., ), coamplification at lower denaturation temperature PCR (COLD‐PCR) (Milbury et al ., ) have been introduced. More recently, digital PCR (dPCR), which uses droplets to compartmentalize individual DNA strands, reached the high sensitivity ranging from 0.1% to 0.001% and is therefore beneficial to detect low allele frequency variants (Gorgannezhad et al ., ; Vogelstein and Kinzler, ).…”

Section: Current Technologies In Ctdnamentioning

confidence: 99%

Liquid biopsy in pancreatic ductal adenocarcinoma: current status of circulating tumor cells and circulating tumor DNA

et al. 2019

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Reliable biomarkers are required to evaluate and manage pancreatic ductal adenocarcinoma. Circulating tumor cells and circulating tumor DNA are shed into blood and can be relatively easily obtained from minimally invasive liquid biopsies for serial assays and characterization, thereby providing a unique potential for early diagnosis, forecasting disease prognosis, and monitoring of therapeutic response. In this review, we provide an overview of current technologies used to detect circulating tumor cells and circulating tumor DNA and describe recent advances regarding the multiple clinical applications of liquid biopsy in pancreatic ductal adenocarcinoma.

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Section: Current Technologies In Ctdnamentioning

confidence: 99%

Liquid biopsy in pancreatic ductal adenocarcinoma: current status of circulating tumor cells and circulating tumor DNA

et al. 2019

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“…10 In melanoma, the level of BRAF V600E cfDNA is useful to monitor disease progression. [11][12][13] To our knowledge, there are no studies of the analysis of cfDNA in patients with EMPD. The aim of this study was to measure cfDNA levels and to assess their clinical significance in EMPD.…”

Section: What Does This Study Add?mentioning

confidence: 99%

“…Although the cfDNA concentration is not significantly different from normal in patients with scleroderma, circulating α1(I) collagen DNA levels are elevated . In melanoma, the level of BRAF V600E cfDNA is useful to monitor disease progression …”

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confidence: 99%

Serum cell‐free DNA levels are a useful marker for extramammary Paget disease

et al. 2019

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Summary Background Although carcinoembryonic antigen (CEA) and cytokeratin 19 fragment (CYFRA) are useful markers for extramammary Paget disease (EMPD), serum CEA and CYFRA levels are not elevated in most patients with EMPD without metastasis. Cell‐free (cf)DNA has attracted attention as an indicator of clinical conditions in several cancers. Objectives To identify further useful biomarkers for the detection of EMPD, including early lesions, and to study the clinical implications of cfDNA in EMPD. Methods cfDNA were isolated from serum of patients with EMPD with and without metastasis, and from healthy volunteers. Serum extracts were amplified using polymerase chain reaction. Results Serum cfDNA levels were significantly elevated in patients with EMPD with or without metastasis compared with those in healthy controls. Serum cfDNA was a better diagnostic marker for the presence of EMPD than serum CYFRA. Moreover, the postoperative serum cfDNA levels were significantly lower than those from the preoperative samples, and the change in serum cfDNA levels reflected the clinical courses of patients with EMPD treated with chemotherapy. Conclusions Taking the evidence together, serum cfDNA levels may be a useful marker for diagnosis and disease progression in EMPD. What's already known about this topic? Serum levels of carcinoembryonic antigen (CEA) and cytokeratin 19 fragment (CYFRA) are not elevated in most patients with extramammary Paget disease (EMPD) without metastasis. Cell‐free (cf)DNA has attracted attention as an indicator of clinical conditions in several cancers. There are few reports of the clinical implications of cfDNA in dermatology. What does this study add? Serum cfDNA levels were significantly elevated in patients with EMPD with or without metastasis compared with those in healthy controls. Postoperative serum cfDNA levels were significantly lower than those from the preoperative samples. Changes in serum cfDNA levels reflected the clinical courses of patients with EMPD treated with chemotherapy. What is the translational message? Serum cfDNA levels in patients with EMPD are a useful marker for the detection of EMPD, including localized EMPD. Changes in serum cfDNA levels in an individual patient may reflect the clinical course of EMPD.

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“…The use of allele-specific blockers to suppress the amplification of the wild-type allele was also chosen for the screening of BRAF mutant alleles in plasma via competitive allele-specific PCR (CastPCR) [25]. However, the analytical sensitivity was lower at 0.5%.…”

Section: Technical Strategies For Ctdna Detection and Quantificationmentioning

confidence: 99%

Plasma Circulating Tumor DNA Levels for the Monitoring of Melanoma Patients: Landscape of Available Technologies and Clinical Applications

Busser

Lupo

Sancey

et al. 2017

BioMed Research International

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Melanoma is a cutaneous cancer with an increasing worldwide prevalence and high mortality due to unresectable or metastatic stages. Mutations in BRAF, NRAS, or KIT are present in more than 60% of melanoma cases, but a useful blood-based biomarker for the clinical monitoring of melanoma patients is still lacking. Thus, the analysis of circulating tumor cells (CTCs) and/or cell-free circulating tumor DNA (ctDNA) analysis from blood (liquid biopsies) appears to be a promising noninvasive, repeatable, and systemic sampling tool for detecting and monitoring melanoma. Here, we review the molecular biology-based strategies used for ctDNA quantification in melanoma patients, as well as their main clinical applications. Droplet digital PCR (ddPCR) and next generation sequencing (NGS) technologies appear to be two versatile and complementary strategies to study rare variant mutations for the detection and monitoring of melanoma progression. Among the different clinical uses of ctDNA, we highlight the assessment of molecular heterogeneity and the identification of genetic determinants for targeted therapy as well as the analysis of acquired resistance. Importantly, ctDNA quantification might also be a novel biomarker with a prognostic value for melanoma patients.

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Quantitative analysis of the BRAF V600E mutation in circulating tumor-derived DNA in melanoma patients using competitive allele-specific TaqMan PCR

Abstract: The castPCR assay can detect and quantitate small amounts of BRAF ctDNA in samples containing large amounts of BRAF cell-free DNA. Thus, we suggest that the castPCR assay is suitable for monitoring ctDNA in the plasma of melanoma patients.

Cited by 35 publications

References 25 publications

Liquid biopsy in pancreatic ductal adenocarcinoma: current status of circulating tumor cells and circulating tumor DNA

Liquid biopsy in pancreatic ductal adenocarcinoma: current status of circulating tumor cells and circulating tumor DNA

Serum cell‐free DNA levels are a useful marker for extramammary Paget disease

Plasma Circulating Tumor DNA Levels for the Monitoring of Melanoma Patients: Landscape of Available Technologies and Clinical Applications

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