Abstract:
BACKGROUND:We explored the utility of multiplexed temperature gradient capillary electrophoresis (TGCE) as a screening tool for identifying genetic changes in the human mitochondrial genome. We examined changes in mitochondrial DNA (mtDNA) in nonmelanoma skin cancers (NMSCs), using TGCE to resolve genetic differences contained within the tumors compared with the control DNA. METHODS:The entire mtDNA from NMSC tissue samples was amplified in 17 overlapping amplicons averaging 1.1 kb in size. Fourteen of these amplicons were digested with restriction endonucleases into as many as five smaller analyzable fragments. Digested tumor mtDNA amplicons were annealed with digested amplicons from the control DNA to form heteroduplexes in regions of DNA mismatch. TGCE was performed in a 96-well parallel format to detect mtDNA changes in a high-throughput fashion. RESULTS:TGCE resolved heteroduplexes from homoduplexes in singlet reactions and in multiplexed assays. Using a single programmed temperature gradient, we detected 18 of 20 mtDNA changes contained within the specimens. This system was also able to detect a single nucleotide change in a fragment as large as 2 kb. CONCLUSION:Multiplexed TGCE is a sensitive and high-throughput screening tool for identifying mtDNA variations.
journal_name
Clin Chemjournal_title
Clinical chemistryauthors
Girald-Rosa W,Vleugels RA,Musiek AC,Sligh JEdoi
10.1373/clinchem.2004.040311subject
Has Abstractpub_date
2005-02-01 00:00:00pages
305-11issue
2eissn
0009-9147issn
1530-8561pii
clinchem.2004.040311journal_volume
51pub_type
杂志文章abstract::The values of apolipoproteins (apo) A-I and B were determined in a population sample of hospital outpatients with a standardized method to verify if the cutpoints calculated in a cross-sectional study in the US are usable with other populations. We also tested the apolipoproteins' ability to discriminate between healt...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1998-01-01 00:00:00
abstract::We evaluated the homogeneous enzyme immunoassay ("EMIT") of serum thyroxine with use of an ABA- 100 Bichromatic Analyzer, modified with an auxiliary dispenser syringe and a dual probe assembly. The modification allows two reagents to be dispensed at one time. The sera must be pre-treated with 0.1 mol/liter NaOH. The p...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1978-01-01 00:00:00
abstract::Crystalline preparations of diiodotyrosine contain trace quantities of thyroxine that can be detected by several competitive radioassays and by chromatography on Sephadex. Spontaneous coupling of diiodotyrosine to thyroxine during iodination of tyrosine and coprecipitation of hormone with diiodotyrosine crystals are l...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1978-10-01 00:00:00
abstract::We evaluated a new kinetic assay for alpha-amylase (Phadebas IsoAmylase Test), using modified starch as the substrate and a CentrifiChem 400 centrifugal analyzer system. We determined isoamylase activities by using a selective inhibitor. Results were compared with those obtained with the chromogenic Phadebas dyed-star...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1983-06-01 00:00:00
abstract::We assessed the LKB "Delfia" (time-resolved dissociation-enhanced lanthanide fluoroimmunoassay) and the Amersham "Amerlite" (enhanced luminescent immunometry) assays of thyrotropin in serum. Both assays are sensitive (respective detection limits: 0.02 and 0.04 milli-int. unit/L) and have very good within- and between-...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1987-08-01 00:00:00
abstract:BACKGROUND:High-sensitivity C-reactive protein (hsCRP) testing is relatively inexpensive and has been shown to predict the risk of cardiovascular disease (CVD) and diabetes in multiple patient groups, including those treated with statin therapy. JUPITER (Justification for the Use of Statins in Primary Prevention: An In...
journal_title:Clinical chemistry
pub_type: 杂志文章,评审
doi:10.1373/clinchem.2008.109728
更新日期:2009-02-01 00:00:00
abstract::We describe a method used for determining serotonin and tryptamine in urine after oral loading with their precursor amino acid. Several factors affecting prepurification and chromatographic separation of both indoleamines were optimized, including sample extraction and mobile-phase composition. Under optimal condition...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1989-01-01 00:00:00
abstract:BACKGROUND:Measurement of serum thyroglobulin (Tg) is used to monitor patients after treatment for differentiated thyroid carcinoma (TC). Difficulty in using Tg as a biomarker of the recurrence of TC in many patients stems from the presence of endogenous anti-Tg autoantibodies (Tg-AAbs), which can interfere with immuno...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2012.195594
更新日期:2013-06-01 00:00:00
abstract::We describe an improved separation of the isoenzymes of aspartate aminotransferase (EC 2.6.1.1), based on ion-exchange chromatography. Involving the "Fast Protein Liquid Chromatography" system (Pharmacia) with a MonoQ column, this rapid, reproducible method for quantifying the mitochondrial enzyme shows good resolutio...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1990-02-01 00:00:00
abstract:BACKGROUND:The term "transrenal DNA" was coined in 2000 to signify that DNA in urine may come from the passage of plasma DNA through the kidney barrier. Although DNA in the urine has the potential to provide a completely noninvasive source of nucleic acids for molecular diagnosis, its existence remains controversial. ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2008.113530
更新日期:2009-04-01 00:00:00
abstract::I describe a gas-chromatographic procedure involving a nitrogen detector for "free" (or "unbound") phenytoin in the concentration range 0.2-4.0 mg/L in ultrafiltrates of plasma. The CV (n = 10) for the assay was 2.9% for a 0.49 mg/L concentration, 1.5% for 2.02 mg/L. The sensitivity was 0.05 mg/L. The ultrafiltration ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1981-03-01 00:00:00
abstract::We present a case of advanced ovarian carcinoma associated with high concentrations of serum and urine amylase. Biochemical analysis to elucidate the nature and origin of the hyperamylasemia identified seven amylase isoenzymes, of which five appeared to be distinct from normal salivary and pancreatic amylase. We discu...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1979-06-01 00:00:00
abstract::A monoclonal-antibody-based competitive radioimmunoassay for measuring human protein C is reported. With use of a purified protein C standard, the solid-phase assay was sensitive to less than 80 micrograms of protein C per liter. Intraassay CVs ranged from 5% to 8%; the inter-assay CV was 5.4%. Analytical recovery ave...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1988-02-01 00:00:00
abstract:BACKGROUND:Plasma creatinine concentration and calculated creatinine clearance are of limited value as glomerular filtration rate (GFR) markers in patients with decompensated liver cirrhosis. We assessed plasma cystatin C as an indicator of GFR in such patients. METHODS:We studied 36 patients with decompensated liver ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:2002-06-01 00:00:00
abstract::Six different procedures for radioiodination of mouse epidermal growth factor (EGF) all resulted in a heterogeneous 125I-labeled EGF preparation, as analyzed by reversed-phase HPLC. EGF preparations that had been iodinated with Chloramine T, lodogen, or lodo-beads were found mainly to consist of oxidized 125I-labeled ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1992-05-01 00:00:00
abstract::We describe modifications to two cartridges, those for alkaline phosphatase and creatinine, on an 18-channel SMAC (Sequential Multichannel Analyzer Computerized), which allow all 18 tests to be run with a commonly prediluted sample. This decreases the required sample size and eliminates the problem of fibrin clots in ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1980-05-01 00:00:00
abstract::This one-dimensional thin-layer chromatographic method is used for assay of phospholipids in the gastric aspirate of newborns. The solvent mixture (chloroform/hexane/methanol/glacial acetic acid/water, 12/7/4/3/0.3 by vol) completely resolves lecithin, sphingomyelin, phosphatidylinositol, phosphatidylserine, phosphati...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1988-04-01 00:00:00
abstract::Plasma hydroxyproline fractions--free-plus-peptide and free--were determined by a method involving centrifugal ultrafiltration and the procedure of Kivirikko et al. [Anal. Biochem. 19, 249 (1967)]. Within- and between-day precision (CV) for the combined fractions was 4--5% and 2.8--6%, respectively. Mean analytical re...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1978-02-01 00:00:00
abstract:BACKGROUND:The lead mobilization test reflects the mobilizable and likely toxicologically active fraction of the lead body burden. We propose a safe and convenient protocol for this test, to assess concomitant copper and zinc excretion and to determine the size of the chelatable lead pool in nonoccupationally exposed a...
journal_title:Clinical chemistry
pub_type: 临床试验,杂志文章
doi:10.1373/clinchem.2005.051128
更新日期:2006-01-01 00:00:00
abstract::Recent evidence indicates that high-density lipoprotein triglyceride (HDL-Tg) may be a predictor of coronary artery disease. We examined three methods for HDL-Tg measurement, comparing results obtained by measurement of Tg in the supernate after heparin-manganese chloride (heparin-Mn) precipitation of EDTA-treated pla...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1988-10-01 00:00:00
abstract:BACKGROUND:Molecular analysis of plasma DNA during human pregnancy has led to the discovery that maternal plasma contains both fetal and maternal DNA. This valuable source of fetal DNA opens up new possibilities for noninvasive prenatal diagnosis. APPROACH:Published data from the last 3 years demonstrating the feasibi...
journal_title:Clinical chemistry
pub_type: 杂志文章,评审
doi:
更新日期:2000-12-01 00:00:00
abstract::Two families with benign hyperphosphatasemia of intestinal origin were studied and compared with six other cases reported in the literature. No evidence of clinical abnormalities or explanations for the unusual enzyme concentrations were found. Agarose gel electrophoresis of alkaline phosphatase (ALP, EC 3.1.3.1) isoe...
journal_title:Clinical chemistry
pub_type: 杂志文章,评审
doi:
更新日期:1991-08-01 00:00:00
abstract:BACKGROUND:The Dutch project "Calibration 2000" aims at harmonization of laboratory results via calibration by development of matrix-based secondary reference materials. We considered the selection, preparation, and characterization of 34 potential reference materials (PRMs). METHODS:Sixteen PRMs were prepared either ...
journal_title:Clinical chemistry
pub_type: 杂志文章,多中心研究
doi:
更新日期:2002-09-01 00:00:00
abstract::To study the effect of fatty acid chain saturation on the fluorescence polarization assay as a measure of fetal lung maturity, we used purified phospholipids isolated from human amniotic fluid and various commercial phospholipids. We found that the fluorescence polarization value decreased as the concentration of unsa...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1986-03-01 00:00:00
abstract::To evaluate a theoretical model for calculating free thyroid hormones, based on the law of mass action, we compared values for both calculated and measured free thyroxin and free triiodothyronine in a group of normal subjects. To determine whether the concentrations of circulating free hormones were also predictable f...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1982-06-01 00:00:00
abstract:BACKGROUND:Standardization of clinical measurements of C-reactive protein (CRP) is based on the availability of reference materials. The serum protein reference material ERM-DA470 was used as a calibrant for various commercially available immunoassays but has now been exhausted. The recently released ERM-DA470k/IFCC wa...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2010.147124
更新日期:2010-09-01 00:00:00
abstract:BACKGROUND:The accurate and precise measurement of urinary albumin is critical, since even minor increases are diagnostically sensitive indicators of renal disease, cardiovascular events, and risk for death. To gain insights into potential measurement biases, we systematically compared urine albumin measurements perfor...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2008.107508
更新日期:2008-09-01 00:00:00
abstract::We compared cotinine, carboxyhemoglobin, and thiocyanate concentrations in blood sampled from 187 cigarette smokers and 181 non-smokers. All three differed significantly between smokers and non-smokers. Cotinine performed best as a test for assessing smoking status, with a sensitivity of 98% as compared with 94% for c...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1984-08-01 00:00:00
abstract:BACKGROUND:Recurrent pregnancy loss, (RPL) affecting 1%-2% of couples, is defined as ≥3 consecutive pregnancy losses before 20-week' gestation. Women with RPL are routinely screened for etiological factors, but routine screening of male partners is not currently recommended. Recently it has been suggested that sperm qu...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2018.289348
更新日期:2019-01-01 00:00:00
abstract:BACKGROUND:Lectin-like oxidized LDL receptor 1 (LOX-1) is implicated in atherothrombotic diseases. Activation of LOX-1 in humans can be evaluated by use of the LOX index, obtained by multiplying the circulating concentration of LOX-1 ligands containing apolipoprotein B (LAB) times that of the soluble form of LOX-1 (sLO...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2009.140707
更新日期:2010-04-01 00:00:00