Abstract:
OBJECTIVES:Defining the BRAF mutation status in metastatic melanoma patients is critical to selecting patients for therapeutic treatment with targeted therapies. Circulating tumour cells (CTCs) can provide an alternative source of contemporaneous tumour genetic material. However methodologies to analyse the presence of rare mutations in a background of wild-type DNA requires a detailed assessment. Here we evaluate the sensitivity of two technologies for cancer mutation detection and the suitability of whole genome amplified DNA as a template for the detection of BRAF-V600 mutations. DESIGN AND METHODS:Serial dilutions of mutant BRAF-V600E DNA in wild-type DNA were tested using both competitive allele-specific PCR (castPCR) and droplet digital PCR (ddPCR), with and without previous whole genome amplification (WGA). Using immunomagnetic beads, we partially enriched CTCs from blood obtained from metastatic melanoma patients with confirmed BRAF mutation positive tumours and extracted RNA and DNA from the CTCs. We used RT-PCR of RNA to confirm the presence of melanoma cells in the CTC fraction then the DNAs of CTC positive fractions were WGA and tested for BRAF V600E or V600K mutations by ddPCRs. RESULTS:WGA DNA produced lower than expected fractional abundances by castPCR analysis but not by ddPCR. Moreover, ddPCR was found to be 200 times more sensitive than castPCR and in combination with WGA produced the most concordant results, with a limit of detection of 0.0005%. BRAF-V600E or V600K mutated DNA was detected in 77% and 44%, respectively, of enriched CTC fractions from metastatic melanoma patients carrying the corresponding mutations. CONCLUSIONS:Our results demonstrate that using ddPCR in combination with WGA DNA allows the detection with high sensitivity of cancer mutations in partially enriched CTC fractions.
journal_name
Clin Biochemjournal_title
Clinical biochemistryauthors
Reid AL,Freeman JB,Millward M,Ziman M,Gray ESdoi
10.1016/j.clinbiochem.2014.12.007subject
Has Abstractpub_date
2015-10-01 00:00:00pages
999-1002issue
15eissn
0009-9120issn
1873-2933pii
S0009-9120(14)00790-5journal_volume
48pub_type
杂志文章abstract::The influence of age and disease on serum albumin and alpha-1-acid glycoprotein was studied in 126 subjects (69 M, 57 F) presenting with tuberculosis and after recovery. Albumin concentration fell with age in both disease and healthy states, and alpha-1-acid glycoprotein concentration rose with age in the healthy stat...
journal_title:Clinical biochemistry
pub_type: 临床试验,杂志文章
doi:10.1016/0009-9120(94)90032-9
更新日期:1994-08-01 00:00:00
abstract:OBJECTIVES:To characterize a novel deletion of exon 3 of CFTR gene and to evaluate the implications in Cystic Fibrosis (CF) care and genetic counseling. DESIGN AND METHODS:We performed a wide mutational analysis of CFTR gene, using reverse dot blot, Multiplex Ligation-dependent Probe Amplification (MLPA) assay and Rea...
journal_title:Clinical biochemistry
pub_type: 杂志文章
doi:10.1016/j.clinbiochem.2011.03.140
更新日期:2011-07-01 00:00:00
abstract::1. The following proteolytic enzymes were measured in muscles of control subjects and patients with muscular dystrophies and related neuromuscular diseases: an elastase-like enzyme, carboxypeptidase A, carboxypeptidase B and pyroglutamyl peptidase. 2. Elastase-like enzyme and carboxypeptidase B did not show significan...
journal_title:Clinical biochemistry
pub_type: 杂志文章
doi:10.1016/s0009-9120(79)80001-6
更新日期:1979-04-01 00:00:00
abstract:OBJECTIVES:Measurements of myoglobin and creatine kinase (CK)-MB isoforms have been suggested to be sensitive tests for the early diagnosis of myocardial infarction (MI). We have investigated the utility of myoglobin, creatine kinase (CK)-MB isoforms and creatine kinase MB mass (CK-MBm) in early diagnosis of MI using c...
journal_title:Clinical biochemistry
pub_type: 杂志文章
doi:10.1016/s0009-9120(02)00385-5
更新日期:2002-11-01 00:00:00
abstract:OBJECTIVES:To measure serum cholinesterase (SCHE) with an integration strategy. DESIGN AND METHODS:At 54.0 micromol/L butyrylthiolcholine, SCHE initial rates were calculated with 50.0 micromol/L butyrylthiolcholine and maximal rates via an improved integrated method if substrate consumptions within 5.0 min were over 6...
journal_title:Clinical biochemistry
pub_type: 杂志文章
doi:10.1016/j.clinbiochem.2008.11.016
更新日期:2009-06-01 00:00:00
abstract::MicroRNAs are endogenous non-coding RNAs that post-transcriptionally regulate gene expression by specifically binding the target mRNA and by consequently inducing its degradation. miRNAs can be released into the circulation where they remain stable and they can be measured. Their changes reflect individual biologic ad...
journal_title:Clinical biochemistry
pub_type: 杂志文章,评审
doi:10.1016/j.clinbiochem.2016.09.017
更新日期:2016-12-01 00:00:00
abstract:OBJECTIVE:MicroRNA-331 (miR-331) has shown regulatory activity against several genes whose expression has been claimed to be deregulated in breast tumors, including that of epidermal growth factor receptor 2 (HER2). Herein, the clinical value of miR-331 expression was investigated by analyzing its levels in breast beni...
journal_title:Clinical biochemistry
pub_type: 杂志文章
doi:10.1016/j.clinbiochem.2018.07.008
更新日期:2018-09-01 00:00:00
abstract::Cyclosporine A (CsA) represents one of the more important therapeutic advances in the field of kidney transplantation. However, its effectiveness is limited by serious side effects, most notably nephrotoxicity. Investigation of the mechanisms of CsA-induced renal dysfunction has been hampered by the lack of a suitable...
journal_title:Clinical biochemistry
pub_type: 杂志文章,评审
doi:10.1016/0009-9120(91)90357-k
更新日期:1991-02-01 00:00:00
abstract:OBJECTIVES:Oxidative stress is considered to be a unifying link between diabetes mellitus (DM) and its complications including nephropathy. The aim of the present study was to evaluate oxidative stress status in Asian Indian patients of type 2 DM with nephropathy. DESIGN AND METHODS:Serum levels of malondialdehyde (MD...
journal_title:Clinical biochemistry
pub_type: 杂志文章
doi:10.1016/s0009-9120(03)00094-8
更新日期:2003-10-01 00:00:00
abstract::This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. ...
journal_title:Clinical biochemistry
pub_type: 杂志文章
doi:10.1016/j.clinbiochem.2006.03.003
更新日期:2006-04-27 00:00:00
abstract:OBJECTIVE:Plasma ammonia measurement is greatly influenced by pre-analytical conditions, which may lead to false positives. We wanted to evaluate the prevalence and clinical impact of plasma ammonia false positives in a pediatric population. DESIGN AND METHODS:Over a 28-month period, charts of patients with elevated a...
journal_title:Clinical biochemistry
pub_type: 杂志文章
doi:10.1016/j.clinbiochem.2007.01.024
更新日期:2007-05-01 00:00:00
abstract:OBJECTIVES:We investigated the relationships of biomarkers of various pathophysiologic pathways including high-sensitivity C-reactive protein (hs-CRP), lipocalin-2 (LCN2), myeloperoxidase (MPO) and matrix metalloproteinases 9 (MMP9) with mortality in stroke patients. DESIGN AND METHODS:hs-CRP, LCN2 and MPO concentrati...
journal_title:Clinical biochemistry
pub_type: 杂志文章
doi:10.1016/j.clinbiochem.2012.06.014
更新日期:2012-11-01 00:00:00
abstract::The toxicity of carbon monoxide has been recognized for long throughout history and is unquestionably the leading cause of unintentional poisoning deaths in the Western countries. The severity of poisoning is dependent upon environmental and human factor. The leading pathophysiological mechanism resides in the ability...
journal_title:Clinical biochemistry
pub_type: 杂志文章,评审
doi:10.1016/j.clinbiochem.2012.06.004
更新日期:2012-11-01 00:00:00
abstract:OBJECTIVE:Protein glutathionylation is considered an important post-translational modification in the pathogenesis of complex diseases. The aim of this study was to examine whether hemoglobin (Hb) is modified by reduced glutathione (GSH) via oxidation of the thiol groups present in diabetes and its associated microangi...
journal_title:Clinical biochemistry
pub_type: 杂志文章
doi:10.1016/j.clinbiochem.2005.06.009
更新日期:2005-10-01 00:00:00
abstract::We present data comparing the Roche CEA radioimmunoassay (RIA) utilizing ultrafiltration and the Abbott CEA immunoradiometric assay (IRMA) methods for standard curve sensitivity, analytical reproducibility, recovery (pre and post extraction), dilution linearity, and patient correlation. The Roche intra- and inter-assa...
journal_title:Clinical biochemistry
pub_type: 杂志文章
doi:10.1016/s0009-9120(82)97007-2
更新日期:1982-10-01 00:00:00
abstract:OBJECTIVES:An overview of biochemical markers of bone metabolism is presented along with indications for their clinical utilization. DESIGN AND METHODS:The structure, cyclical metabolism, and hormone regulation of bone is reflected by markers of resorption, formation and/or turnover. Markers of resorption representing...
journal_title:Clinical biochemistry
pub_type: 杂志文章,评审
doi:10.1016/s0009-9120(97)00113-6
更新日期:1997-12-01 00:00:00
abstract:OBJECTIVE:To develop a Rapid Flow-through assay for distinguishing replicating Tiantan vaccine-generated serological response from true HIV infection. DESIGN AND METHOD:A Rapid Flow-through Test including gp41, gp36, sk1, sk2 and sk3 antigens was established and the performance of the assay was evaluated in clinical s...
journal_title:Clinical biochemistry
pub_type: 杂志文章
doi:10.1016/j.clinbiochem.2012.05.013
更新日期:2012-10-01 00:00:00
abstract:OBJECTIVES:Current extraction methods for cobalamins from serum influence the molecular characteristics of the vitamin. Therefore, an extraction procedure that leaves the cobalamins unchanged is needed. DESIGN AND METHODS:Monoclonal antibodies towards transcobalamin (TC) and haptocorrin (HC) (in house) linked to magne...
journal_title:Clinical biochemistry
pub_type: 杂志文章
doi:10.1016/j.clinbiochem.2007.09.008
更新日期:2007-12-01 00:00:00
abstract::Retrospective analysis of DNA from paraffin-embedded fixed bone marrow biopsy specimens is possible if preceded by amplification of the DNA sequences of interest by the polymerase chain reaction (PCR). These fixed specimens yield degraded DNA that may not be suitable for direct analysis by conventional digestion and h...
journal_title:Clinical biochemistry
pub_type: 杂志文章
doi:10.1016/0009-9120(92)80051-h
更新日期:1992-04-01 00:00:00
abstract:BACKGROUND:Polycyclic aromatic hydrocarbons (PAHs), some ubiquitous environmental contaminants are capable to cause oxidative stress, during its metabolism. It is believed that many diseases that have a common origin in oxidative stress begin in childhood. Considering oxidative stress evolved during PAHs metabolism as ...
journal_title:Clinical biochemistry
pub_type: 杂志文章
doi:10.1016/j.clinbiochem.2007.11.017
更新日期:2008-02-01 00:00:00
abstract::1,5-anhydroglucitol (AG) is a nonmetabolizable glucose analogue found in plasma due to ingestion. The normal steady-state concentration can be dramatically decreased by inhibition of tubular reabsorption during periods of hyperglycemia. For this reason, monitoring of AG has been plausibly advocated for detection of pe...
journal_title:Clinical biochemistry
pub_type: 杂志文章,评审
doi:10.1016/j.clinbiochem.2008.08.086
更新日期:2009-02-01 00:00:00
abstract:BACKGROUND:The availability of reference intervals is essential for physicians to interpret laboratory results. Most of our laboratory reference intervals are derived from data on the foreign population. We have studied the reference intervals of 24 common laboratory biochemical tests in an apparently healthy adult Han...
journal_title:Clinical biochemistry
pub_type: 杂志文章
doi:10.1016/j.clinbiochem.2012.06.022
更新日期:2012-10-01 00:00:00
abstract:OBJECTIVES:To ascertain the effects of eating on plasma antioxidant capacity in patients with liver disease. DESIGN AND METHODS:Eighteen cirrhotic patients were compared to 18 age and sex-matched controls. TRAP was measured by a fluorometric assay after a 12 h fast, and 60, 120, and 180 min after the study participant...
journal_title:Clinical biochemistry
pub_type: 杂志文章
doi:10.1016/s0009-9120(99)00039-9
更新日期:1999-08-01 00:00:00
abstract::The serum argino-succinate lyase (ASAL) activity was measured in preoperative and post-operative serum samples, from approximately 500 patients submitted to elective surgery. The results were compared with the determinations of 6 other enzyme activities on the same serum specimens. Serum ASAL elevations correlated hig...
journal_title:Clinical biochemistry
pub_type: 杂志文章
doi:10.1016/s0009-9120(75)92144-x
更新日期:1975-06-01 00:00:00
abstract:OBJECTIVES:We emphasize the importance of routine follow-up of subnormal TSH concentrations with QC materials. DESIGN AND METHODS:The functional sensitivity (FS) of the ADVIA Centaur system TSH assay was assessed. We report the values yielded for QC materials in two clinical laboratories. RESULTS:The FS was <0.02 mIU...
journal_title:Clinical biochemistry
pub_type: 杂志文章
doi:10.1016/j.clinbiochem.2012.04.017
更新日期:2012-10-01 00:00:00
abstract:BACKGROUND:Disease-independent sources of biomarker variability include pre-analytical, analytical and biological variance. The aim of the present study was to evaluate whether the pre-analytical phase has any impact on the emerging heart disease TWEAK and HMGB1 protein markers and miRNA biomarkers, and whether peptido...
journal_title:Clinical biochemistry
pub_type: 杂志文章
doi:10.1016/j.clinbiochem.2016.11.005
更新日期:2017-03-01 00:00:00
abstract:OBJECTIVES:The ratio of glycated albumin to HbA1c (GA/HbA1c ratio) is a known indicator that reflects fluctuations in plasma glucose. In this study, the association of the GA/HbA1c ratio to diabetic nephropathy and diabetic retinopathy in patients with type 2 diabetes was investigated. DESIGN AND METHODS:Among patient...
journal_title:Clinical biochemistry
pub_type: 杂志文章
doi:10.1016/j.clinbiochem.2016.11.032
更新日期:2017-04-01 00:00:00
abstract:OBJECTIVES:The aim of this study was to investigate the diagnostic utility of plasma neutrophil gelatinase-associated lipocalin (NGAL) as an early objective biomarker to predict acute kidney injury (AKI) in critically ill patients with suspected sepsis, for whom procalcitonin (PCT) was used for the diagnosis and stagin...
journal_title:Clinical biochemistry
pub_type: 杂志文章
doi:10.1016/j.clinbiochem.2013.05.069
更新日期:2013-10-01 00:00:00
abstract:OBJECTIVES:The newly developed glomerular filtration rate (GFR)-estimating equations developed by the CKD-EPI Collaboration and Feng et al. (2013) that are based on standardized serum cystatin C (ScysC), combined/not combined with serum creatinine (Scr), require further validation in China. We compared the performance ...
journal_title:Clinical biochemistry
pub_type: 杂志文章
doi:10.1016/j.clinbiochem.2014.05.060
更新日期:2014-09-01 00:00:00
abstract:OBJECTIVES:The aim of the study was to examine the long-term effect of commonly used whole soy foods (soy flour) and purified daidzein (one major isoflavone and the precursor of equol) on renal function among prehypertensive postmenopausal women who are also equol producers, a population most likely to benefit from soy...
journal_title:Clinical biochemistry
pub_type: 临床试验,杂志文章,随机对照试验
doi:10.1016/j.clinbiochem.2014.05.054
更新日期:2014-09-01 00:00:00