Abstract：
:We combined the amplification refractory mutation system (ARMS) and fluorescence polarization (FP) to give a homogeneous genomic DNA genotype analysis method. Oligonucleotide probes labeled with the fluorescein dyes fluorescein isothiocyanate and 5-([4,6-dichlorotriazin-2-yl]amino)fluorescein and the rhodamine dye 6-carboxyrhodamine were included in amplification mixes and were annealed to PCR products after amplification. Hybridization was accompanied by an increase in the FP of the probe. We demonstrated homogeneous genotyping by analyzing human DNA samples for delta F508 mutation status of the cystic fibrosis transmembrane conductance regulator gene. The genotypes determined with the method described herein were in full agreement with those obtained by the conventional application of ARMS. We also demonstrated the simultaneous detection of two PCR products in a single reaction. The assay method described is homogeneous and so obviates the necessity to open reaction vessels after amplification. This therefore eliminates PCR carryover contamination.
journal_name
Clin Chemjournal_title
Clinical chemistryauthors
Gibson NJ,Gillard HL,Whitcombe D,Ferrie RM,Newton CR,Little Ssubject
Has Abstractpub_date
1997-08-01 00:00:00pages
1336-41issue
8 Pt 1eissn
0009-9147issn
1530-8561journal_volume
43pub_type
杂志文章abstract：:To compare six assays of hyaluronan (hyaluronic acid; HYA) in serum, developed in different laboratories, we analyzed 10 samples from each of three groups: healthy persons, patients with primary biliary cirrhosis, and patients with rheumatoid arthritis. All the assays are based on the use of affinity proteins specific...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1992-01-01 00:00:00
abstract：:We developed a fluorometric method for determining delta-aminolevulinic acid (ALA) in urine of lead workers. A high-performance liquid chromatograph (HPLC) equipped with a fluorescence HPLC monitor is used. The detection limit for aqueous ALA is 20 micrograms/L (15 pmol of ALA in the 100-microL sample). The working li...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1987-09-01 00:00:00
abstract：:We measured the free amino acids in plasma of 58 patients with HIV infection and in six persons in the risk group. The HIV+ patients had significantly increased concentrations of arginine, phenylalanine, and glutamate in comparison with both age- and sex-matched controls and the members of the risk group. Glutamate co...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1988-09-01 00:00:00
abstract：BACKGROUND:Uremic syndrome is the consequence of the retention of solutes usually cleared by the healthy kidneys. p-Cresol can be considered a prototypic protein-bound uremic toxin. It is conceivable, analogous with drugs, that the non-protein-bound fraction of p-cresol exerts toxicity. This aspect had never been evalu...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/49.3.470
更新日期：2003-03-01 00:00:00
abstract：:A rapid technique (5-10 min) has been developed for fractionating nucleotides from base and nucleoside contaminants in acid extracts of cells, by adsorption to silica gels. Silica gels (1-mL bed volume) were washed with 5 mL of water then with 5 mL of acetonitrile/water (90/10 by vol). After applying 3-mL samples, adj...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1980-09-01 00:00:00
abstract：BACKGROUND:The TDx Fetal Lung Maturity II (FLM II) assay uses amniotic fluid to assess lung maturity of the unborn infant. We investigated common preanalytical factors that influence FLM results, including centrifugation, sample storage, and contamination by whole blood. METHODS:We tested 18 specimens after centrifuga...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/49.6.935
更新日期：2003-06-01 00:00:00
abstract：BACKGROUND:Recent reports have described inherent problems with androgen immunoassays compared with mass spectrometry analyses. METHODS:We developed a method for measuring serum testosterone (T) and 5alpha-dihydrotestosterone (DHT) simultaneously via liquid-liquid extraction followed by liquid chromatography-tandem ma...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2008.103846
更新日期：2008-11-01 00:00:00
abstract：:Ammonia concentrations in plasma may increase because of contamination and deterioration of blood components during specimen handling and storage. Using replicate specimens from healthy volunteers, we studied influences of specimen processing and storage procedures on ammonia measurements made with a self-contained re...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1984-06-01 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：:A new method (enzyme-ligand immunoassay, ELIA) is described for the estimation of estrogen (ER) and progesterone (PR) receptors in microsamples of human breast cancer tissue. The technique, based on the nonisotopic measurement of receptor-bound estradiol and progesterone, involves three steps: (a) simultaneous saturat...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1995-01-01 00:00:00
abstract：BACKGROUND:Mitochondrial diseases are clinically and genetically heterogeneous, with variable penetrance, expressivity, and differing age of onset. Disease-causing point mutations and large deletions in the mitochondrial genome often exist in a heteroplasmic state. Current molecular analyses require multiple different ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2011.181438
更新日期：2012-09-01 00:00:00
abstract：:The carboxyl terminal region of corticotropin (ACTH), (Ala34-Phe-Pro-Glu-Leu-Phe39), a region of the hormone conserved during evolution, served as an antigen for the production of a sequence-specific antiserum. In a radioimmunoassay, peptides that extend toward the amino terminal from Ala34, such as [Tyr,Gly]-ACTH34-3...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1981-04-01 00:00:00
abstract：:A high (2.0 mol/L) NaCl concentration apparently suppresses the secondary-site binding of bilirubin by human serum albumin. Thus if an excess of bilirubin is added to human serum albumin or to neonatal serum in buffer containing 0.1 mol of tris(hydroxymethyl)aminomethane and 2.0 mol of NaCl per liter (pH 7.5), and the...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1980-01-01 00:00:00
abstract：BACKGROUND:The sex differential in coronary heart disease (CHD) risk, which is not explained by male/female differences in lipid and lipoprotein concentrations, narrows with age. We examined whether this differential CHD risk might, in part, be attributable to the sizes of lipoprotein particles or concentrations of lip...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2004.032763
更新日期：2004-07-01 00:00:00
abstract：:To validate azotemic markers as an index for intradialytic changes in solute concentration, we compared eight solutes (pseudouridine, xanthine, hypoxanthine, peak 4, peak 5, p-hydroxyhippuric acid, indoxyl sulfate, and hippuric acid) with five classical azotemic markers (urea, creatinine, uric acid, phosphate, and pot...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1992-08-01 00:00:00
abstract：:We compared the performance of an automated assay of creatine kinase MB isoenzyme (CK-MB) mass (Stratus) with that of a CK-MB enzymatic assay routinely used at our institutions. Both of these assays use the same CK-MB-specific monoclonal antibody to immunocapture CK-MB, thus providing a direct means of comparing a mas...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1989-10-01 00:00:00
abstract：:We have developed a simple, specific, and sensitive reversed-phase liquid-chromatographic method for accurate and simultaneous analysis of corticosterone and cortisol in human plasma. We achieved a detection limit of 300 ng/L for both steroids by modifying the old solid-phase extraction method to make use of "Tef Elut...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1992-03-01 00:00:00
abstract：BACKGROUND:The thiopurine S:-methyltransferase (TPMT) genetic polymorphism has a significant clinical impact on the toxicity of thiopurine drugs, which are used in the treatment of leukemia and as immunosuppressants. To date, 10 mutant alleles are known that are associated with intermediate or low TPMT activity. To fac...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：2001-03-01 00:00:00
abstract：BACKGROUND:Hyperglycosylated human chorionic gonadotropin (hCG-h) contains larger and more complex carbohydrate chains than regular human chorionic gonadotropin (hCG). hCG-h is thought to be the major form of hCG produced by testicular cancers and it has been suggested to play a key role in tumor invasion, but studies ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2012.183723
更新日期：2012-07-01 00:00:00
abstract：:We describe a new method for quantitatively measuring substances of clinical interest by high-performance liquid affinity chromatography (HPLAC). As a model system we selected analysis for transferrin in human serum with immobilized antibodies in a high-performance liquid chromatographic system. SelectiSpher-10 Activa...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1988-10-01 00:00:00
abstract：:We modified the liquid-chromatographic assay of Schleicher and Wieland (J Clin Chem Clin Biochem 1981; 19:81-7) for measuring lysine-bound glucose in serum proteins, increasing its performance and practicality. After precipitating serum proteins from 10- to 50-microL samples with ethanol (700 mL/L) and hydrolyzing the...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1987-09-01 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：:Of 98 patients' specimens examined for alkaline phosphatase (EC 3.1.3.1) isoenzymes by electrophoresis on cellulose acetate membrane after incubation with neuraminidase, 50 showed only a single liver or bone isoenzyme staining band; in 15 of these, the tissue origin of the fraction could not be accurately identified f...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1985-07-01 00:00:00
abstract：:The Dutch Foundation for Quality Assessment in Clinical Chemistry (SKZL) is the professional organization that conducts external quality-assessment schemes in The Netherlands. However, such schemes in fact assess the performance of the internal quality-control systems of the participating laboratories. In this paper w...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1991-07-01 00:00:00
abstract：:We evaluated a new discrete analyzer, the Hitachi 705. The instrument can process sequentially as many as 19 different enzymes or other analytes, with a throughput of 180 tests per hour. We evaluated the spectrophotometer, the sample and reagent probes, and the performance for 11 assays: creatine kinase, aspartate ami...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1983-04-01 00:00:00
abstract：:We have developed and thoroughly validated radioimmunoassays (RIAs) for osteocalcin in plasma and serum, demonstrating independence of volume and determining the recovery of added standard and within- and between-assay precision. Antibodies were raised in rabbits and mice by immunization with either osteocalcin adsorb...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1989-07-01 00:00:00
abstract：:We studied the effect of lyophilization of serum pools on the determination of cholesterol with the enzymatic methods used in five automated analyzers: SMAC (Technicon), the RA-1000 (Technicon), the aca (Du Pont), the TDx (Abbott), and the Ektachem 700P (Kodak). We prepared two serum pools: pool A (2.2 g/L) and pool B...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1989-07-01 00:00:00
abstract：BACKGROUND:Plasma amino acid (PAA) profiles are used in routine clinical practice for the diagnosis and monitoring of inherited disorders of amino acid metabolism, organic acidemias, and urea cycle defects. Interpretation of PAA profiles is complex and requires substantial training and expertise to perform. Given previ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1093/clinchem/hvaa134
更新日期：2020-09-01 00:00:00
abstract：:We determined the LD-1/LD-2 isoenzyme ratio in hemolysates of erythrocytes by electrophoresis on cellulose acetate and on agarose. A ratio exceeding 1.0 was found with the former but not the latter. Results were similar for in vitro models of hemolytic disorders. Using cellulose acetate electrophoresis, we determined ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1981-08-01 00:00:00
abstract：:We describe a method for measuring urinary oxalate and citrate with capillary electrophoresis (CE) and indirect ultraviolet absorbance detection. Sample preparation is minimal, requiring an acidification, brief centrifugation, and dilution. The method is rapid, with oxalate and citrate having mean migration times of 4...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1995-09-01 00:00:00
