Abstract：
BACKGROUND:Screening for heterozygous sequence changes in PCR products, also known as "mutation scanning", is an important tool for genetic research and clinical applications. Conventional methods require a separation step. METHODS:We evaluated the sensitivity and specificity of homogeneous scanning, using a saturating DNA dye and high-resolution melting. Heterozygous single-nucleotide polymorphism (SNP) detection was studied in three different sequence backgrounds of 40%, 50%, and 60% GC content. PCR products of 50-1000 bp were generated in the presence of LCGreen I. After fluorescence normalization and temperature overlay, melting curve shape was used to judge the presence or absence of heterozygotes among 1632 cases. RESULTS:For PCR products of 300 bp or less, all 280 heterozygous and 296 wild-type cases were correctly called without error. In 672 cases between 400 and 1000 bp with the mutation centered, the sensitivity and specificity were 96.1% and 99.4%, respectively. When the sequence background and product size with the greatest error rate were used, the sensitivity of off-center SNPs (384 cases) was 95.6% with a specificity of 99.4%. Most false negatives occurred with SNPs that were compared with an A or T wild type sequence. CONCLUSIONS:High-resolution melting analysis with the dye LCGreen I identifies heterozygous single-base changes in PCR products with a sensitivity and specificity comparable or superior to nonhomogeneous techniques. The error rate of scanning depends on the PCR product size and the type of base change, but not on the position of the SNP. The technique requires only PCR reagents, the dye LCGreen I, and 1-2 min of closed-tube, post-PCR analysis.
journal_name
Clin Chemjournal_title
Clinical chemistryauthors
Reed GH,Wittwer CTdoi
10.1373/clinchem.2003.029751subject
Has Abstractpub_date
2004-10-01 00:00:00pages
1748-54issue
10eissn
0009-9147issn
1530-8561pii
clinchem.2003.029751journal_volume
50pub_type
杂志文章abstract：:A method for determining platinum in biological materials by flameless atomic absorption spectrometry has been developed for use in pharmacological studies of platinum antitumor compounds. Tissue digests are analyzed in a heated graphite atomizer, with use of background correction and ramp temperature charring. We hav...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1977-07-01 00:00:00
abstract：:The kidney and its response to the antidiuretic hormone (ADH) are the principal protective mechanisms necessary to maintain a normal plasma tonicity (osmolality). Hence, determination of the response of the ADH-renal axis to an abnormal plasma tonicity is an important step to understanding water homeostasis. Determina...
journal_title：Clinical chemistry
pub_type： 杂志文章,评审
doi：
更新日期：1994-07-01 00:00:00
abstract：:To correct the results of a radioimmunoassay for beta-thyrotropin (TSH) subunit by eliminating the proportion ascribable to intact TSH, we have devised a method that experimentally reproduces the conditions under which the interference develops. Beta-TSH subunit was assayed in several preparations containing known con...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1987-05-01 00:00:00
abstract：:We describe an automated (ABA-100) enzymic method for determination of urinary oxalate by use of oxalate oxidase (EC 1.2.3.4) isolated from beet stems. The H2O2 produced by the oxidation of oxalate by oxalate oxidase is measured by coupling with oxidation and conjugation of 3-methyl-3-benzothiazolinone hydrazone with ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1983-10-01 00:00:00
abstract：:The deleterious, disruptive effects of estrogen mimics on the endocrine system were discovered after the compounds were released into the environment. Their chemical structure does not obviously resemble that of steroid hormones; hence, their estrogenic effects were totally unexpected. In addition to occupational expo...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1995-12-01 00:00:00
abstract：:In this efficient extraction and isocratic liquid-chromatographic procedure for measuring eight antidepressant drugs in serum (amoxapine, 8-hydroxyamoxapine, doxepin, desmethyldoxepin, imipramine, desipramine, amitriptyline, and nortriptyline), they are extracted from serum as a group by use of disposable solid-phase ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1984-11-01 00:00:00
abstract：BACKGROUND:It has been shown that a high percentage of interleukin-8 (IL-8) in blood is cell associated. Recently, a simple method for determination of cell-associated IL-8 in whole blood after cell lysis has been described. The purpose of this study was to evaluate this method, to examine the influence of preanalytic ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：2000-09-01 00:00:00
abstract：:A modified gas-liquid chromatographic method for determining plasma concentrations of bupivacaine and lidocaine is described, with cyclizine as an internal standard. The extraction procedure requires no solvent evaporation, thus overcoming the problem of drug volatility. Concentrations as low as 0.1 mg/liter can be de...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1978-09-01 00:00:00
abstract：:Estimation of plasma catecholamine concentrations by "high-performance" liquid chromatography, although increasingly used, still presents difficulties. We describe a method for clean-up of plasma samples on cation-exchange resin before they are extracted with alumina. This procedure eliminates many of the interfering ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1983-07-01 00:00:00
abstract：:We evaluated a new agarose-gel-electrophoretic procedure (Corning) (I) for separating and quantitating of high-density lipoprotein cholesterol (HDLC), comparing it with the modified Lipid Research Clinics (LRC) procedure (heparin 183 kilounits/L, MnCl2 92 mmol/L) (II). Method I was insensitive to an HDLC concentration...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1979-11-01 00:00:00
abstract：BACKGROUND:We evaluated kinetic changes of high-sensitivity cardiac troponin T (hs-cTnT) in patients with acute coronary syndrome (ACS) and patients with hs-cTnT increases not due to ACS to rule in or rule out non-ST-segment elevation myocardial infarction (STEMI). METHODS:hs-cTnT was measured serially in consecutive ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2011.171827
更新日期：2012-01-01 00:00:00
abstract：:Diminished ability to utilize acetate (acetate intolerance) developed in a male patient on chronic hemodialysis after five years of maintenance dialysis. His ability to utilize lactate was also subnormal. We studied acetate metabolism in vitro by isolating lymphocytes from the patient's blood before dialysis and measu...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1979-03-01 00:00:00
abstract：:Serum free thyroxin (FT4) was determined in 40 patients with various nonthyroidal illnesses. We studied seven methods: (1) a free thyroxin index calculated from total T4 and triiodothyronine resin uptake; (2) a free T4 index determined by enzyme inhibitor assays (Abbott's "Tetrazyme" and "Thyrozyme"); (3) a free T4 in...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1983-12-01 00:00:00
abstract：BACKGROUND:The determination of cellular beta-galactocerebrosidase activity is an established procedure to diagnose Krabbe disease and monitor the efficacy of gene/stem cell-based therapeutic approaches aimed at restoring defective enzymatic activity in patients or disease models. Current biochemical assays for beta-ga...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2008.115873
更新日期：2009-03-01 00:00:00
abstract：BACKGROUND:B-type natriuretic peptide (BNP), which is activated in heart failure (HF), is processed to an active form by corin. The corin gene is expressed in the human heart and kidney, but corin protein expression in the heart, kidney, and circulation, along with whether proBNP is processed by circulating corin, rema...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2010.153908
更新日期：2011-01-01 00:00:00
abstract：:We describe an enzyme-linked immunosorbent assay (ELISA) for urinary albumin. It requires only commercially available reagents, can detect as little as 16 micrograms of albumin per liter, and analytical recovery ranges from 92 to 116%. The assay is simple, rapid, and inexpensive. Albumin excretion was 6.2 (SD 4.1) mg/...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1987-04-01 00:00:00
abstract：:We evaluated the Hybritech Tandem-E procedure for quantifying choriogonadotropin (hCG) in human serum. In this "sandwich"-type assay, two monoclonal antibodies directed against different regions of the hCG molecule are used, one coated on a plastic bead, the second conjugated to alkaline phosphatase. The assay can det...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1985-03-01 00:00:00
abstract：BACKGROUND:Prostate-specific antigen (PSA) is an important marker for the diagnosis and management of prostate cancer, and the free PSA/total PSA ratio has been shown to be efficient for distinguishing prostate cancer from benign prostatic hyperplasia. We report here the characterization of seven mouse monoclonal antib...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1999-05-01 00:00:00
abstract：:Serial measurements of the bone and fetal intestinal isoenzymes of alkaline phosphatase (EC 3.1.3.1) in the plasma of 43 term and 43 preterm infants, from birth to six weeks later, indicate that the bone isoenzyme gradually increases over this period in both preterm and term infants fed with unsupplemented commercial ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1987-10-01 00:00:00
abstract：BACKGROUND:Concerns exist regarding how the 99th percentile upper reference limit (URL) of cardiac troponin (cTn) is determined and whether it should be derived from normal healthy individuals. CONTENT:The 99th percentile URL of cTn is an important criterion to standardize the diagnosis of myocardial infarction (MI) f...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1093/clinchem/hvaa158
更新日期：2020-09-01 00:00:00
abstract：BACKGROUND:Some of the genetic variants of hemoglobin (Hb) and their chemically modified species are known to affect the measurement of Hb A(1c). The purpose of this study was to characterize Hb species in the blood sample of a 74-year-old German male with an exceptionally low Hb A(1c) value. METHODS:Hemolysates from ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/49.1.137
更新日期：2003-01-01 00:00:00
abstract：BACKGROUND:Extracellular vesicles (EVs) contain a rich cargo of different RNA species with specialized functions and clinical applications. However, the landscape and characteristics of extracellular vesicle long RNA (exLR) in human blood remain largely unknown. METHODS:We presented an optimized strategy for exLR sequ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2018.301291
更新日期：2019-06-01 00:00:00
abstract：:This new one-step chemiluminescent immunoassay of free thyroxin (FT4) involving a thyroxin-immunoglobulin conjugate labeled with acridinium ester (Magic Lite System; Ciba Corning Diagnostics Corp., Medfield, MA) is rapid (one 1-h incubation), requires two calibrators per run, and takes 10 s per sample for the quantifi...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1988-12-01 00:00:00
abstract：:In "flow-injection analysis," the sample is inserted into a stream of reagent by use of a sample-injection valve. Mixing occurs downstream from the valve in a coil of tubing. With chemiluminescence detection this coil is positioned in front of a photomultiplier. We have evaluated the system for detection of hydrogen p...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1979-09-01 00:00:00
abstract：BACKGROUND:Meconium analysis is a diagnostically sensitive and objective alternative to maternal self-report for detecting prenatal tobacco exposure. Nicotine and metabolite disposition in meconium is poorly characterized, and correlation of analytes' concentrations with neonatal outcomes is unexplored. Our objectives ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2008.109173
更新日期：2008-12-01 00:00:00
abstract：BACKGROUND:Formation of deposits of the insoluble amyloid beta-peptide is believed to be causally related with neurodegeneration in Alzheimer disease (AD). The beta-peptide originates from a larger amyloid precursor protein (APP) by the action of proteolytic enzymes. The first proteolytic event leading to amyloid forma...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2006.066720
更新日期：2006-06-01 00:00:00
abstract：BACKGROUND:The multiple endocrine neoplasia 2 (MEN 2) syndromes [MEN 2A, MEN 2B, and familial medullary thyroid carcinoma (FMTC)] are caused by germline mutations of the RET protooncogene. Because 85% of MEN 2A patients and 30% of FMTC patients have mutations at codon 634, the recommended molecular analyses begin at ex...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：2001-11-01 00:00:00
abstract：BACKGROUND:This Case Conference reviews the normal changes in thyroid activity that occur during pregnancy and the proper use of laboratory tests for the diagnosis of thyroid dysfunction in the pregnant patient. CASE:A woman in the 18th week of pregnancy presented with tachycardia, increased blood pressure, severe vom...
journal_title：Clinical chemistry
pub_type： 杂志文章,评审
doi：
更新日期：1999-12-01 00:00:00
abstract：BACKGROUND:Reduced renal function is an established risk factor for cardiovascular events. We compared 3 measures of renal function--serum cystatin C, serum creatinine, and calculated creatinine clearance--as predictors of subsequent cardiovascular events in a community-based population of elderly individuals. METHODS...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2009.135962
更新日期：2010-05-01 00:00:00
abstract：:Radiometric, colorimetric, and two immunochemical methods for measuring total iron-binding capacity are compared. We evaluated the procedures on the basis of precision, applicability to a pediatric population, and accuracy as assessed by analytical recovery of purified transferrin. The immunoephelometric assay for tra...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1978-10-01 00:00:00
