Abstract：
BACKGROUND:False-positive results are a common problem in real-time PCR identification of DNA sequences that differ from near neighbors by a single-nucleotide polymorphism (SNP) or deletion. Because of a lack of sufficient probe specificity, post-PCR analysis, such as a melting curve, is often required for mutation differentiation. METHODS:Tentacle Probes, cooperative reagents with both a capture and a detection probe based on specific cell-targeting principles, were developed as a replacement for 2 chromosomal TaqMan-minor groove binder (MGB) assays previously developed for Yersinia pestis and Bacillus anthracis detection. We compared TaqMan-MGB probes to Tentacle Probes for SNP and deletion detection based on the presence or absence of a growth curve. RESULTS:With the TaqMan-MGB Y. pestis yp48 assays, false-positive results for Yersinia pseudotuberculosis occurred at every concentration tested, and with the TaqMan-MGB B. anthracis gyrA assays, false-positive results occurred in 21 of 29 boil preps of environmental samples of near neighbors. With Tentacle Probes no false-positive results occurred. CONCLUSIONS:The high specificity exhibited by Tentacle Probes may eliminate melting curve analysis for SNP and deletion mutation detection, allowing the diagnostic use of previously difficult targets.
journal_name
Clin Chemjournal_title
Clinical chemistryauthors
Satterfield BC,Kulesh DA,Norwood DA,Wasieloski LP Jr,Caplan MR,West JAdoi
10.1373/clinchem.2007.091488subject
Has Abstractpub_date
2007-12-01 00:00:00pages
2042-50issue
12eissn
0009-9147issn
1530-8561pii
clinchem.2007.091488journal_volume
53pub_type
杂志文章abstract：:We describe specifications for high-purity 4-nitrophenol, which is suitable for spectrophotometric standardization. Such a reference material is needed in clinical enzymology to establish the proper molar absorptivity of 4-nitrophenol under final reaction conditions, particularly for measuring alkaline phosphatase act...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1980-05-01 00:00:00
abstract：:Solid-phase-based immunoassays have traditionally been plagued by nonspecific binding to the solid phase and by slow reaction kinetics relative to reactants that are free to diffuse in solution. We have developed two novel immunoassays in which the solid phase is generated in situ after the specific binding reaction h...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1987-09-01 00:00:00
abstract：BACKGROUND:24,25-Dihydroxyvitamin D [24,25(OH)2D] in serum may be both a nuisance and nutritionally valuable. METHODS:We investigated the impact of 24,25(OH)2D3 on the performance of commercially available immunoassays for serum total 25-hydroxyvitamin D [25(OH)D] using (a) serum from a nationally representative sampl...
journal_title：Clinical chemistry
pub_type： 杂志文章,随机对照试验
doi：10.1373/clinchem.2014.234955
更新日期：2015-04-01 00:00:00
abstract：:A new type of multilayer film analytical element--consisting of a spreading layer, a blocking layer, an enzymic reagent layer, and a transparent layer--has been developed for blood glucose determination. After a spot of whole blood is dropped on the film, the plasma glucose concentration is determined in 6 min without...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1981-07-01 00:00:00
abstract：:We present an improved fructosamine assay for use with the Technicon RA-1000 analyzer. Features of this assay include: specimen throughput of 100 per hour, within-batch CV of 1.5%, between-batch CV of 2.4%, standard curve linear up to 24.9 mmol of fructosamine per liter, and good correlation (r = 0.82) with hemoglobin...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1985-12-01 00:00:00
abstract：:We evaluated superoxide dismutase (EC 1.15.1.1) and glutathione peroxidase (EC 1.11.1.9) activities in the erythrocytes of lead-exposed and nonexposed workers from distinct industries localized in the State of São Paulo, Brazil. Concentrations of either lead or protoporphyrin IX, or both, in blood were used to indicat...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1985-10-01 00:00:00
abstract：:We describe a highly accurate and precise method for determination of total cholesterol in serum by isotope dilution/mass spectrometry. The method was developed for a Study Group of the Committee on Standards of the American Association for Clinical Chemistry, for use in establishing the accuracy of a candidate refere...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1980-06-01 00:00:00
abstract：:We describe a completely automated enzymic system for measuring total cholesterol in serum. All reagents are contained in an analytical test pack and the test is performed on Du Pont's Automatic Clinical Analyzer (aca), which mixes the sample (20 microliter) and reagents and performs the necessary absorbance measureme...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1978-01-01 00:00:00
abstract：BACKGROUND:The new Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equation was developed to address the systematic underestimation of the glomerular filtration rate (GFR) by the Modification of Diet in Renal Disease (MDRD) Study equation in patients with a relatively well-preserved kidney function. The per...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2009.135111
更新日期：2010-03-01 00:00:00
abstract：BACKGROUND:The promise of targeted therapies in molecularly defined subsets of cancer has led to a transformation of the process of drug development in oncology. To target cancer successfully and precisely requires high-quality translational data. Such data can be generated by the use of biomarkers that answer key ques...
journal_title：Clinical chemistry
pub_type： 杂志文章,评审
doi：10.1373/clinchem.2012.191072
更新日期：2013-01-01 00:00:00
abstract：BACKGROUND:The recent discovery of the presence of circulating cell-free fetal DNA in maternal plasma opens up new prenatal diagnostic applications and provides new avenues for clinical investigation. It is of research and potential diagnostic interest to determine whether fetal trisomy 21 may be associated with quanti...
journal_title：Clinical chemistry
pub_type： 临床试验,杂志文章,多中心研究
doi：
更新日期：1999-10-01 00:00:00
abstract：:A modified heparin-Sepharose affinity chromatography procedure (Boberg et al., J. Lipid Res. 18:544-547, 1977) was developed to determine two different triglyceride lipase activities in human post-heparin plasma: hepatic triglyceride lipase (I) and lipoprotein lipase (II). With this procedure, lipoproteins were separa...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1981-05-01 00:00:00
abstract：BACKGROUND:The use of nonstandardized N-terminal pro-B-type natriuretic peptide (NT-proBNP) assays can contribute to the misdiagnosis of heart failure (HF). Moreover, there is yet to be established a common consensus regarding the circulating forms of NT-proBNP being used in current assays. We aimed to characterize and...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2012.200204
更新日期：2013-10-01 00:00:00
abstract：:Creatine kinase (CK) isoenzymes MM, MB, and BB are located primarily in the cell cytosol, and increased CKMB in plasma is the hallmark of myocardial infarction. However, whether CK is released with reversible ischemic injury remains controversial. Here, we assessed plasma CK activity--cytosolic and mitochondrial CK--i...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1997-03-01 00:00:00
abstract：:We describe a new double-antibody enzyme radioimmunoassay for thyroxin-binding globulin (TBG) in serum. A TBG-beta-D-galactosidase complex was prepared with m-maleimidobenzoyl N-hydroxysuccinimide ester. The mean analytical recovery of TBG added to serum was 99%, sample volume and TBG value were linearly related, and ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1982-12-01 00:00:00
abstract：:A convenient screening procedure for presence of drugs of abuse in urine consists of two steps: adsorption of the drugs from urine onto a paper loaded with cation-exchange resin and detection of the adsorbed drugs by direct radioimmunoassay. The first step can be performed in the field, the second in a central laborat...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1977-10-01 00:00:00
abstract：:Backpropagation neural networks are a computer-based pattern-recognition method that has been applied to the interpretation of clinical data. Unlike rule-based pattern recognition, backpropagation networks learn by being repetitively trained with examples of the patterns to be differentiated. We describe and analyze t...
journal_title：Clinical chemistry
pub_type： 临床试验,杂志文章,随机对照试验
doi：
更新日期：1993-09-01 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: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：BACKGROUND:PCR-based detection of minimal residual disease (MRD) in neuroblastoma (NB) patients can be used for initial staging and monitoring therapy response in bone marrow (BM) and peripheral blood (PB). PHOX2B has been identified as a sensitive and specific MRD marker; however, its expression varies between tumors....
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2008.117945
更新日期：2009-07-01 00:00:00
abstract：:We measured the concentration of erythrocyte protoporphyrin in 43 blood samples by two fluorometric methods. In the indirect method, protoporphyrin was extracted from blood and the fluorescence of the free acid in the extract was determined with a filter-fluorometer. These results were compared with results of direct ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1978-09-01 00:00:00
abstract：:We describe the sensitive and precise determination 5-hydroxytryptophan in plasma from patients treated with this compound. The method is based on solvent extraction of plasma, high-performance liquid chromatography and continuous fluorometric detection after reaction with phthaldialdehyde in strong acid. The combined...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1978-02-01 00:00:00
abstract：:We describe radioimmunoassay for human prostatic acid phosphatase [orthophosphoric-monoester phospho-hydrolase (acid optimum), EC 3.1.3.2] in serum, with use of monospecific antisera raised in rabbits against highly purified acid phosphatase from human prostates. The antiserum did not cross react with partly purified ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1978-11-01 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：:Cholinesterase activity in human serum was measured calorimetrically and compared to values obtained with the spectrophotometric method of Rappaport et al. [Clin. Chim. Acta 4, 227 (1959)]. The response of the calorimeter upon mixing serum with acetylcholine corresponded to the cholinesterase activity of the serum, th...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1977-10-01 00:00:00
abstract：BACKGROUND:In patient and population samples, generation of analytical results that are comparable and independent of the measurement system, time, and location is essential for the utility of laboratory information supplied in healthcare. Obtaining analytical measurement results with such characteristics is the aim of...
journal_title：Clinical chemistry
pub_type： 杂志文章,评审
doi：10.1373/clinchem.2008.107052
更新日期：2009-06-01 00:00:00
abstract：:Humans are exposed daily to low concentrations of many different chemical substances, natural and some man-made. Although many of these substances can be toxic at high levels, typical exposures are far below the effect levels. The responses produced by man-made aromatic hydrocarbon receptor agonists, such as dioxins, ...
journal_title：Clinical chemistry
pub_type： 杂志文章,评审
doi：
更新日期：1996-08-01 00:00:00
abstract：:We describe a sensitive beta-endorphin (beta-EP) radioimmunoassay specific for beta-EP 1-31 applied to human ventricular and lumbar cerebrospinal fluid (CSF). Specificity was documented by reversed-phase HPLC of CSF pools. Simultaneous ventricular and lumbar CSF samples from 13 patients suspected of having normal-pres...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1992-06-01 00:00:00
abstract：:Synovial fluid samples were collected from 45 patients with rheumatoid arthritis, spondylarthropathy, or osteoarthritis, to study their content of elastase (EC 3.4.21.37) and of cysteine proteinases (EC 3.4.22.1, 3.4.22.15). We measured both elastase complexed with alpha 1-proteinase inhibitor and elastase activity to...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1992-09-01 00:00:00
abstract：:Abnormal creatine kinase (CK) isoenzyme patterns were observed in the serum of a 64-year-old woman with severe heart disease. Agarose electrophoresis revealed the presence of all the usual CK isoenzymes (MM, MB, and BB) plus an extra band between MM and MB. Total serum CK activity was within the normal range. Within 2...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1982-10-01 00:00:00
