Abstract:
BACKGROUND:PCR-based mutation detection is prone to methodological errors, e.g., in restriction length fragment polymorphism (RFLP) and allele-specific amplification (ASA), false PCR results may occur because of technical faults or atypical new mutations. METHODS:We investigated the ability of a genotyping assay based on hybridization of labeled oligonucleotides to detect and discriminate known and as yet unknown mutations in the factor V and apolipoprotein B-100 genes. Expected melting points were calculated using a nearest-neighbor model for nucleic acid duplex stability and compared with experimental findings derived from LightCycler melting curves. A method for genotyping the apolipoprotein B-100 G10699A and C10698T mutations is presented. RESULTS:All mismatches tested for in the probed sequence could be detected with a single probe. The measured melting points were in good agreement with their values predicted using the nearest-neighbor model (r = 0.96; y = 0.98x + 1.18; S(y|x) = 0.96; n = 24). CONCLUSIONS:This procedure not only allows the identification of the mutation of interest but also enables the discrimination from other potential mutations in the vicinity of the former. The nearest-neighbor model is valid for hybridization probe assays on the LightCycler and should be of general value in setting up such assays. We have shown for two clinically relevant genotyping examples that the LightCycler mutation detection system has superior discriminatory performance compared with conventional RFLP or ASA PCR-based methods for molecular diagnostic purposes. With this method, in every hybridization probe assay, all mutations under a properly designed probe should be detectable, but they will not necessarily be discriminated from each other in all cases.
journal_name
Clin Chemjournal_title
Clinical chemistryauthors
von Ahsen N,Oellerich M,Armstrong VW,Schütz Esubject
Has Abstractpub_date
1999-12-01 00:00:00pages
2094-101issue
12eissn
0009-9147issn
1530-8561journal_volume
45pub_type
杂志文章abstract:BACKGROUND:Because cyclosporin A (CsA) and glucocorticoids inhibit the production of interleukin-2 (IL-2) and other cytokines, quantitative analysis of cytokine mRNA might constitute a pharmacodynamic measure for immunosuppressive drug effects. We investigated whether immunosuppressive drugs influence cytokine mRNA exp...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:2002-12-01 00:00:00
abstract::This method for assaying digoxin in serum with improved specificity combines small-column extraction of serum, "high-performance" liquid chromatography, and RIA of the eluted fractions. Analytical recoveries of 1.0, 0.5, and 0.1 microgram/L standards were 95%, 93%, and 84%, respectively. The CVs for duplicates and rep...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1985-08-01 00:00:00
abstract::Parathyroid hormone-related protein (PTHrP) was measured in plasma by a new immunoradiometric assay (IRMA) from Nichols Institute. The assay is specific for PTHrP and shows excellent parallelism when measuring keratinocyte fluid, samples with high PTHrP content, and PTHrP-supplemented plasma. A precision profile estab...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1993-03-01 00:00:00
abstract::9-(2-Phosphonylmethoxyethyl)adenine (PMEA), (S)-9-(3-hydroxy-2-phosphonylmethoxypropyl)adenine (HPMPA), and (R,S)-9-(3-fluoro-2-phosphonylmethoxypropyl)adenine (FPMPA) are selective antiviral agents with activity against a broad spectrum of DNA viruses and retroviruses. A highly sensitive method has been developed to ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1992-04-01 00:00:00
abstract:BACKGROUND:Cardiac troponins are specific biochemical markers of myocardial injury used in the diagnosis of acute myocardial disease and cardiac risk stratification. To avoid misclassification of patients, troponin assays must demonstrate precision at the low end of the measuring range. We report our evaluation of the ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2005.057216
更新日期:2006-02-01 00:00:00
abstract::Owing to increased enzymatic hydrolysis of glutamine, additional ammonia is formed in blood and plasma specimens with increased gamma-glutamyltransferase activity (gamma-GT, EC 2.3.2.2.). The close correlation between gamma-GT and glutaminase (EC 3.5.1.2) activities (r = 0.97) in plasma, the inhibition with 6-diazo-5-...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1990-08-01 00:00:00
abstract::We measured glucocorticoid receptors (GR) in mononuclear leukocytes (MNL) isolated from peripheral blood of 145 apparently healthy volunteers (86 men and 59 women). An age-related decrease in the number of GR was suggested between subjects younger than 20 years and elderly subjects; there was no apparent seasonal vari...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1991-10-01 00:00:00
abstract:BACKGROUND:Stability of chemical speciation during sample handling and storage is a prerequisite to obtaining reliable results of trace element speciation analysis. There is no comprehensive information on the stability of common arsenic species, such as inorganic arsenite [As(III)], arsenate [As(V)], monomethylarsonic...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1999-11-01 00:00:00
abstract::We have developed an automated latex agglutination test for Treponema pallidum (TPLA) for measurement of the concentration of treponemal antibodies in syphilitic sera. The assay needs neither a complicated pretreatment of the sera nor special techniques. Intra- and interassay precision studies showed high reproducibil...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1993-08-01 00:00:00
abstract::On-line, simultaneous measurement of blood K+, Ca2+, Na+, and pH has been achieved by using a quadruple-function ChemFET (chemical-sensitive field-effect transistor) integrated-circuit sensor. Blood is withdrawn from the subject through an actively heparinized, dual-lumen cannula and passed into a flow-cell containing...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1984-01-01 00:00:00
abstract::The Glucose-Controlled Insulin Infusion System (Biostator) is a modular, computerized, feedback control system for dynamic control of blood glucose concentrations in diabetics. This on-line glucose analyzer for use with whole blood utilizes a novel enzyme (glucose oxidase)-membrane configuration and an electrochemical...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1978-08-01 00:00:00
abstract:BACKGROUND:Technological advances have resulted in a renaissance of proteomic studies directed at finding markers of disease progression, diagnosis, or responsiveness to therapy. Renal diseases are ideally suited for such research, given that urine is an easily accessible biofluid and its protein content is derived mai...
journal_title:Clinical chemistry
pub_type: 杂志文章,评审
doi:10.1373/clinchem.2011.165969
更新日期:2012-02-01 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 divided patients with brain lesions into three groups: (a) patients with primary or metastatic brain cancer, (b) brain infarctions, and (c) brain contusion(s). We analyzed each patient's sera for creatine kinase isoenzyme BB (CK-BB), using a monoclonal antibody kit (Impres-BB; International Immunoassay Laboratories...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1989-04-01 00:00:00
abstract:BACKGROUND:Proteomic technology permits the investigation of genetic metabolic diseases at the level of protein expression. Changes in the expression, polypeptide structure, and posttranslational modification of individual proteins can be detected in complex mixtures of proteins. METHODS:We used high-resolution two-di...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:2001-11-01 00:00:00
abstract::Ion chromatography is a new analytical technique in which the ions to be analyzed are separated on a special pellicular ion-exchange column. Once separated, the effluent from the separating column passes into a second ion-exchange column, which removes the eluent or buffer ions from the moving phase without removing t...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1976-09-01 00:00:00
abstract::We evaluated six precipitation methods for high-density lipoprotein cholesterol (HDL-chol) determination: the heparin/Mn2+ precipitation reagent method (Hep), two variants of the phosphotungstic acid/Mg2+ method (Tung-L and Tung-B), the dextran sulfate 50,000/Mg2+ method (Dex), the PEG 6000 method (PEG), and the PEG 6...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1997-04-01 00:00:00
abstract::Binary-system capillary electrophoresis (CE) with head-column field-amplified sample stacking permits determination of amiodarone and desethylamiodarone in 20-microL serum samples. The assay is characterized by a detection limit for both compounds of 80 nmol/L and by excellent linear response over the recommended ther...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1996-11-01 00:00:00
abstract::Unsaturated cobalamin-binding capacity in plasma (P-UBBC) is determined by use of a silica gel (QUSO G32) to separate haptocorrin (P-ApoHC) and transcobalamin (P-ApoTC). The method is sensitive and precise: detection limit 13 pmol/L, interassay coefficients of variation 3% for P-UBBC (mean = 1080 pmol/L), 4% for P-Apo...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1989-07-01 00:00:00
abstract::The tricyclic antidepressants amitriptyline, nortriptyline, imipramine, and desipramine in serum of patients taking one of the drugs were quantified in two laboratories by high-performance liquid chromatography (HPLC) and enzyme-multiplied immunoassay (EMIT; Syva). Results for split samples were highly correlated, but...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1988-11-01 00:00:00
abstract::The immunoreactivity of anti-triiodothyronine (anti-T3) IgG, pretreated at acidic pH and adsorbed onto polystyrene beads, was significantly greater than that determined for native anti-T3 IgG immobilized at optimum pH 3.5. Acidic-pH-pretreated IgG, adsorbed at pH 7.0 onto ethanol-treated beads, had less immunoreactivi...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1990-03-01 00:00:00
abstract::We measured three renal tubular brush-border enzymes (lactate dehydrogenase, LDH, EC 1.1.1.27; gamma-glutamyltransferase, GGT, EC 2.3.2.2; and alkaline phosphatase, AP, EC 3.1.3.1) in morning urine samples from 48 healthy subjects to check whether different storage times and temperatures could modify enzyme concentrat...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1991-08-01 00:00:00
abstract::We report a case of hyperphosphatasemia in a 35-year-old patient with hepatitis B who underwent an orthotopic xenogeneic liver transplant. Marked increases in total alkaline phosphatase (ALP; EC 3.1.3.1) activity began 5 days posttransplantation (six times human normal) and increased to approximately 17 times normal a...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1994-07-01 00:00:00
abstract::Currently, we can make the following generalizations about the major long-term complications of diabetes. First, they occur commonly after 10 to 15 years of diabetes. Second, the complications of renal and retinal disease, but not of atherogenic and neurological disease, appear to be related to the severity of hypergl...
journal_title:Clinical chemistry
pub_type: 临床试验,杂志文章,评审
doi:
更新日期:1986-10-01 00:00:00
abstract::We find lag phases exceeding 20 min in measuring creatine kinase activity, by using the kinetic creatin phosphate leads to creatine assay, in sera of some patients with carcinoma metastatic to the liver. Such long lag phases are accompanied by a decreased apparent enzyme activity. These problems are eliminated by addi...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1976-11-01 00:00:00
abstract:BACKGROUND:To allow multianalyte binding assays, we have developed a novel polystyrene microtiter plate containing 24 main wells, each divided into 7 subwells. We explored its clinical potential by developing a PCR-chemiluminescent immunoassay (PCR-CLEIA) for simultaneous detection and typing of seven high oncogenic ri...
journal_title:Clinical chemistry
pub_type: 临床试验,杂志文章
doi:
更新日期:2002-10-01 00:00:00
abstract::A fixed-time incubation method for measuring acetylsalicylic acid esterase was improved by using a higher concentration of the substrate, acetylsalicylic acid, and an activator, Ca2+. The enzymatic activity in serum was 25-fold that measured by earlier methods. Inhibition studies showed a pattern similar to that repor...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1983-03-01 00:00:00
abstract::We report the results of a laboratory and clinical evaluation of a commercial kit procedure (Immuno Nuclear Corp.) for measuring the mid-region of the parathyrin molecule. The estimated dose at 50% binding averaged 130 pmol/L, and the minimum detectable concentration was 14 pmol/L. The within-assay CV was less than or...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1986-11-01 00:00:00
abstract:BACKGROUND:Massively parallel sequencing has recently been used in noninvasive prenatal diagnosis. The current costs of this technology are still relatively expensive, however, and sample throughput is still relatively low when it is used as a molecular diagnostic tool. Rather than nonselectively sequencing the genome,...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2010.154336
更新日期:2011-01-01 00:00:00
abstract:BACKGROUND:The optimal situation in external quality assessment (EQA) is to use commutable materials. No previous study has examined the commutability of a whole-blood material for point-of-care (POC) testing. The aim of this study was to determine the commutability of the Norwegian Quality Improvement of Laboratory Ex...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2018.300202
更新日期:2019-06-01 00:00:00