Abstract:
Objectives:Colorectal cancer (CRC) screening using stool samples is now in routine use where tumor DNA methylation analysis for leading markers such as NDRG4 and SDC2 is an integral part of the test. However, processing stool samples for reproducible and efficient extraction of human genomic DNA remains a bottleneck for further research into better biomarkers and assays. Methods:We systematically evaluated several factors involved in the processing of stool samples and extraction of DNA. These factors include: stool processing (solid and homogenized samples), preparation of DNA from supernatant and pellets, and DNA extraction with column and magnetic beads-based methods. Furthermore, SDC2 and NDRG4 methylation levels were used to evaluate the clinical performance of the optimal protocol. Results:The yield of total and human genomic DNA (hgDNA) was not reproducible when solid stool scraping is used, possibly due to sampling variations. More reproducible results were obtained from homogenized stool samples. Magnetic beads-based DNA extraction using the supernatant from the homogenized stool was chosen for further analysis due to better reproducibility, higher hgDNA yield, lower non-hgDNA background, and the potential for automation. With this protocol, a combination of SDC2 and NDRG4 methylation signals with a linear regression model achieved a sensitivity and specificity of 81.82 and 93.75%, respectively. Conclusions:Through the systematic evaluation of different stool processing and DNA extraction methods, we established a reproducible protocol for analyzing tumor DNA methylation markers in stool samples for colorectal cancer screening.
journal_name
Clin Chem Lab Medjournal_title
Clinical chemistry and laboratory medicineauthors
Jin S,Ye Q,Hong Y,Dai W,Zhang C,Liu W,Guo Y,Zhu D,Zhang Z,Chen S,Wang Y,Li D,Ma W,Yang Z,Li J,Zheng Z,Luan J,Wu X,Jiang F,Xu C,Ding Cdoi
10.1515/cclm-2020-0300subject
Has Abstractpub_date
2020-07-16 00:00:00pages
91-99issue
1eissn
1434-6621issn
1437-4331pii
cclm-2020-0300journal_volume
59pub_type
杂志文章abstract:BACKGROUND:When performed erroneously, the venous blood specimen collection (VBSC) practice steps patient identification, test request management and test tube labeling are at high risk to jeopardize patient safety. VBSC educational programs with the intention to minimize risk of harm to patients are therefore needed. ...
journal_title:Clinical chemistry and laboratory medicine
pub_type: 杂志文章
doi:10.1515/cclm-2018-0051
更新日期:2018-10-25 00:00:00
abstract:BACKGROUND:The biochemical determination of cardiac natriuretic peptides, primarily brain natriuretic peptide (BNP) and the amino-terminal fragment of its pro-hormone proBNP (NT-proBNP), are reliable tools for diagnosing cardiac disease, establishing prognosis and evaluating the effectiveness of treatment. These biomar...
journal_title:Clinical chemistry and laboratory medicine
pub_type: 杂志文章,多中心研究
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更新日期:2010-07-01 00:00:00
abstract:BACKGROUND:Galectin-3 is an emerging biomarker of heart failure and of myocardial fibrosis risk. Monitoring of galectin-3 is essential during treatment with galectin-3 inhibitors. The aim of our study was to assess long-term biological variability in a specific group of unhealthy subjects. METHODS:The biological varia...
journal_title:Clinical chemistry and laboratory medicine
pub_type: 临床试验,杂志文章
doi:10.1515/cclm-2013-1088
更新日期:2015-01-01 00:00:00
abstract::It is not clear whether brain natriuretic peptide (BNP) or N-terminal proBNP (NT-proBNP) is superior as a diagnostic and prognostic indicator in cardiac diseases. Here, we compare the clinical correlations of both peptides in a population of 92 ambulatory patients with heart failure, using a well-established immunorad...
journal_title:Clinical chemistry and laboratory medicine
pub_type: 杂志文章
doi:10.1515/CCLM.2002.130
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abstract:BACKGROUND:The timely detection of severe inflammatory conditions is of great importance for early therapy initiation and the patient's prognosis. The complex humoral and cellular processes involved in inflammation offer many opportunities for diagnostic testing, which are still unused in clinical practice. We investig...
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pub_type: 杂志文章
doi:10.1515/cclm-2014-0482
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abstract::Congenital disorders of glycosylation (CDG) are genetic multisystemic diseases due to various defects in the biosynthesis or processing of glycoproteins. Our aim is to present our experience in the selective screening of CDG syndrome in a paediatric population (421 patients) with clinical suspicion of the disease, ana...
journal_title:Clinical chemistry and laboratory medicine
pub_type: 杂志文章
doi:10.1515/CCLM.2000.143
更新日期:2000-10-01 00:00:00
abstract::Folate measurements, particularly for whole blood, show wide inter-laboratory and inter-methodology variability. This variability appears to be due in part to the lack of internationally accepted reference materials. A whole blood haemolysate, lyophilised in ampoules and designated 95/528, was therefore evaluated by 1...
journal_title:Clinical chemistry and laboratory medicine
pub_type: 杂志文章
doi:10.1515/CCLM.2004.090
更新日期:2004-05-01 00:00:00
abstract:BACKGROUND:There is a certain difference in opinion regarding the optimal choice of housekeeping genes used as normalization factors in gene expression analysis. We have therefore examined the suitability of three housekeeping genes, hypoxanthine phosphoribosyl transferase, beta-glucuronidase and beta2-micro-globulin, ...
journal_title:Clinical chemistry and laboratory medicine
pub_type: 杂志文章
doi:10.1515/CCLM.2007.303
更新日期:2007-01-01 00:00:00
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journal_title:Clinical chemistry and laboratory medicine
pub_type: 杂志文章
doi:10.1515/cclm-2020-0210
更新日期:2020-08-07 00:00:00
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journal_title:Clinical chemistry and laboratory medicine
pub_type: 杂志文章
doi:10.1515/cclm-2012-0610
更新日期:2013-07-01 00:00:00
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journal_title:Clinical chemistry and laboratory medicine
pub_type: 杂志文章
doi:10.1515/CCLM.2002.197
更新日期:2002-11-01 00:00:00
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journal_title:Clinical chemistry and laboratory medicine
pub_type: 杂志文章
doi:10.1515/CCLM.2001.065
更新日期:2001-05-01 00:00:00
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journal_title:Clinical chemistry and laboratory medicine
pub_type: 杂志文章,评审
doi:10.1515/CCLM.2003.194
更新日期:2003-09-01 00:00:00
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journal_title:Clinical chemistry and laboratory medicine
pub_type: 杂志文章
doi:10.1515/CCLM.2000.066
更新日期:2000-05-01 00:00:00
abstract:BACKGROUND:It has been proposed that only anti-thyroid-peroxidase (TPOAb) and not anti-thyroglobulin (TgAb) antibodies should be assayed in the diagnosis of autoimmune thyroid disease (AITD). METHODS:We analyzed the thyroid autoantibody profile with a multiplex bead array system, which allows simultaneous measurement ...
journal_title:Clinical chemistry and laboratory medicine
pub_type: 杂志文章
doi:10.1515/CCLM.2006.137
更新日期:2006-01-01 00:00:00
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journal_title:Clinical chemistry and laboratory medicine
pub_type: 杂志文章
doi:10.1515/CCLM.2007.108
更新日期:2007-01-01 00:00:00
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journal_title:Clinical chemistry and laboratory medicine
pub_type: 杂志文章
doi:10.1515/CCLM.2007.192
更新日期:2007-01-01 00:00:00
abstract::The effects of the time of the day and breakfast on the outcome of interleukin-6 and its soluble receptor in serum were studied on 22 subjectively health volunteers. We collected blood specimens at 8.00 a.m. after an overnight fast, at 9.30 a.m. after usual breakfast, and at 11.30 a.m. A second group of volunteers (n ...
journal_title:Clinical chemistry and laboratory medicine
pub_type: 杂志文章
doi:10.1515/CCLM.1998.054
更新日期:1998-05-01 00:00:00
abstract:BACKGROUND:This paper describes the evolution, structure, operation and some outcomes of the Brazilian Laboratory Indicators Program created by the Brazilian Society of Clinical Pathology/Laboratory Medicine (Sociedade Brasileira de Patologia Clínica/Medicina Laboratorial, or SBPC/ML), in partnership with ControlLab, a...
journal_title:Clinical chemistry and laboratory medicine
pub_type: 杂志文章
doi:10.1515/cclm-2012-0357
更新日期:2012-11-01 00:00:00
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journal_title:Clinical chemistry and laboratory medicine
pub_type: 杂志文章,评审
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pub_type: 杂志文章
doi:10.1515/CCLM.2011.623
更新日期:2011-08-01 00:00:00
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pub_type: 杂志文章
doi:10.1515/CCLM.2009.302
更新日期:2009-01-01 00:00:00
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journal_title:Clinical chemistry and laboratory medicine
pub_type: 杂志文章
doi:10.1515/cclm-2013-0610
更新日期:2014-04-01 00:00:00
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journal_title:Clinical chemistry and laboratory medicine
pub_type: 杂志文章,评审
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更新日期:2004-01-01 00:00:00
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journal_title:Clinical chemistry and laboratory medicine
pub_type: 杂志文章,评审
doi:10.1515/CCLM.2005.197
更新日期:2005-01-01 00:00:00
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journal_title:Clinical chemistry and laboratory medicine
pub_type: 杂志文章
doi:10.1515/cclm-2014-0546
更新日期:2015-02-01 00:00:00
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journal_title:Clinical chemistry and laboratory medicine
pub_type: 杂志文章
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更新日期:2019-05-27 00:00:00
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journal_title:Clinical chemistry and laboratory medicine
pub_type: 杂志文章
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更新日期:2019-04-24 00:00:00
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journal_title:Clinical chemistry and laboratory medicine
pub_type: 杂志文章
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更新日期:2017-03-01 00:00:00
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journal_title:Clinical chemistry and laboratory medicine
pub_type: 杂志文章,评审
doi:10.1515/CCLM.2009.331
更新日期:2009-01-01 00:00:00