Abstract：
:A new method for the reliable identification of localized variations in DNA by detection of associated diagnostic products with matrix-assisted laser desorption ionization time-of-flight mass spectrometry is described. The diagnostic products are generated by the primer oligo base extension (PROBE) reaction, which requires a single detection primer complementary to a region down-stream of a target strand's variable site. On addition of a polymerase, three dNTPs, and the fourth nucleotide in dideoxy form, the primer is extended through the mutation region until the first ddNTP is incorporated; the mass of the extension products determines the composition of the variable site. Tests for five cystic fibrosis mutations, including two exon 11 sites measured in a biplex reaction, and for differentiating between three common alleles of the poly(T) tract at the intron 8 splice acceptor site of the CFTR gene are presented. All experimental steps required for PROBE are amenable to the high degree of automation desirable for a high-through-put diagnostic setting. Furthermore, it requires no fluorescent, chemiluminescent, or radioactive labeling; the mass signals measured offer a far more analytically definitive signal, leading in all cases to high-quality unambiguous and easily interpreted results.
journal_name
Clin Chemjournal_title
Clinical chemistryauthors
Braun A,Little DP,Köster Hsubject
Has Abstractpub_date
1997-07-01 00:00:00pages
1151-8issue
7eissn
0009-9147issn
1530-8561journal_volume
43pub_type
杂志文章abstract：:The ultraviolet fluorescence emission spectra of sera from apparently healthy persons and of sera from cancer patients frequently show significantly different curve shapes. A biparametric fluorescence method has been developed to use these deviations to detect patients with malignant diseases (Clin Chem 1986;32:1974-8...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1990-11-01 00:00:00
abstract：:I evaluated use of the fluorescence polarization technique to measure neocarzinostatin, a proteinaceous antitumor antibiotic, and its antibody, in serum. The antigen (neocarzinostatin), labeled with fluorescein isothiocyanate, was allowed to interact with its antibody in a cuvet, in the instrument, yielding an increas...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1978-12-01 00:00:00
abstract：BACKGROUND:Distinguishing adenocarcinoma and squamous cell carcinoma subtypes of non-small cell lung cancers is critical to patient care. Preoperative minimally-invasive biopsy techniques, such as fine needle aspiration (FNA), are increasingly used for lung cancer diagnosis and subtyping. Yet, histologic distinction of...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1093/clinchem/hvaa207
更新日期：2020-11-01 00:00:00
abstract：:In this rapid, simple enzyme immunoassay for carcinoembryonic antigen (CEA) in serum the "sandwich" system is used. The CEA is first extracted from the serum into acetate buffer by heating at 70 degrees C, then incubated in a polyvinyl microtitration plate previously coated with anti-CEA. The bound CEA is then reacted...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1983-12-01 00:00:00
abstract：BACKGROUND:Bilirubin has antioxidative and cytoprotective properties. Low plasma concentrations of bilirubin are reportedly associated with the development of coronary and cerebrovascular disease, and bilirubin concentrations are strongly correlated with the enzyme activity of the hepatic uridine diphosphate glucuronos...
journal_title：Clinical chemistry
pub_type： 临床试验,杂志文章
doi：10.1373/clinchem.2007.102046
更新日期：2008-05-01 00:00:00
abstract：:We present a unified approach to the use of linear models and matrix least squares with the intention of providing a better understanding of the techniques themselves and of the statistics that arise from these techniques as they are used in clinical chemistry. Emphasis is placed on the importance of appropriate exper...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1979-06-01 00:00:00
abstract：BACKGROUND:The analysis of red cell folate (RCF) depends on complete hemolysis of erythrocytes, and it is assumed that complete hemolysis is achieved by 10-fold dilution of whole blood with hypotonic solutions of 10 g/L ascorbic acid/ascorbate. This report challenges this assumption. METHODS:The conventional method of...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：2000-12-01 00:00:00
abstract：:Optimal reaction conditions for assaying human lactate dehydrogenase pyruvate-to-lactate were determined for isoenzymes 1 and 5 at 25, 30, and 37 degrees C. Three of the nine different buffers examined--imidazole, triethanolamine, and N-tris(hydroxymethyl)-methyl-2-aminoethane sulfonic acid--are satisfactory. Beta-NAD...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1978-02-01 00:00:00
abstract：:Detection of intracellular antigens by flow cytometry requires effective fixation and permeabilization of the cell membrane. This study compares three fixation/permeabilization techniques: two commercial chemical reagents, the ORTHOPermeaFix (OPF) and the FIX&PERM Cell Permeabilization Kit (F&P), and a novel method ba...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1998-11-01 00:00:00
abstract：:We have modified the 63Ni radiochemical method of Ho (Anal Biochem 1970;36:105) for determination of free fatty acids (FFA) in plasma. Extracting 1 or 0.1 mL of plasma with Dole's mixture (J Biol Chem 1960;235:2595) and washing the "heptane" layer with two volumes of isopropanol/water/dilute (0.5 mol/L) H2SO4 (25/25/1...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1987-01-01 00:00:00
abstract：:Human myocardial creatine kinase isoenzyme MM is present as a single form in tissue, but upon its release into plasma two additional forms, with faster anodal migration, are apparent on polyacrylamide electrophoresis. We designate the three forms as MM3, MM2, and MM1 in increasing order of anodal mobility. When tissue...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1984-05-01 00:00:00
abstract：BACKGROUND:Endothelin-1 (ET-1), a potent vasoconstrictor, is difficult to measure because of its instability and its binding to receptors and plasma proteins. We report a rapid, robust way to indirectly quantify ET-1 release by measuring the C-terminal ET-1 precursor fragment (CT-proET-1) without an extraction step. M...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2005.065581
更新日期：2006-06-01 00:00:00
abstract：:We describe the performance of an RIA for the measurement of prostate-specific antigen (PA). Between-assay precision (CV) for control sera with various analyte concentrations was as follows: mean = 1.67 micrograms/L, CV = 7.1%; mean = 4.47 micrograms/L, CV = 5.6%; mean = 7.15 micrograms/L, CV = 5.5% (n = 19 each). Ana...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1984-05-01 00:00:00
abstract：:We have observed an electrophoretically abnormal, nonfunctional species of alpha 1-antitrypsin in serum from patients who were receiving bone-marrow transplants for treatment of leukemia or aplastic anemia. Three of four patients in whose serum this protein appeared died soon after; the fourth recovered, and the disap...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1987-01-01 00:00:00
abstract：:We describe a micro-scale method for determining serum theophylline. The chromatography system includes a muBondapack C18 column and acetonitrile, 70 ml/liter of sodium acetate buffer (10 mmol/liter, ph 4.0) as the mobile phase. Test serum or plasma, 30 mul, is mixed with an equal quantity of a solution containing the...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1977-03-01 00:00:00
abstract：:A method is described for separating antibody-bound from free fractions in ligand assays by column affinity chromatography, and its application to radioimmunoassay of choriomammotropin. In the method, 70 x 10 mm (i.d.) polypropylene columns containing about 150 mg of immunosorbent (goat anti-rabbit gamma-globulins cov...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1981-06-01 00:00:00
abstract：:The concentration of the conjugated bile acid, cholylglycine, in serum is a sensitive and specific indicator of hepatic function. We describe a convenient, specific, and precise radioimmunoassay for cholylglycine, in which 125I-labeled cholylglycyltyrosine is used as tracer. In addition, a blocking agent in the buffer...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1981-10-01 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 studied the specificity of kinetic methods of analysis, with emphasis on creatinine determination. The error contributed by an interferent that reacts with the analytical reagent and absorbs at the wavelength of analysis is shown to be related to the ratio of the equilibrium absorbances of the analyte and interfere...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1980-04-01 00:00:00
abstract：:We describe a sensitive method for the determination of vanadium in biological substances by atomic absorption spectroscopy. By using background correction and pyrolytically coated graphite tubes with flameless apparatus, this vanadium assay has a sensitivity of 65 pg and a detection limit of 30 pg, approximately sixf...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1982-01-01 00:00:00
abstract：BACKGROUND:Institutions face increasing charges related to molecular genetic testing for neurological diseases. The literature contains little information on the utilization and performance of these tests. METHODS:A retrospective utilization review was performed to determine the diagnostic yield of neurogenetic tests ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2006.083360
更新日期：2007-06-01 00:00:00
abstract：:A modified pentamer formation assay (PFA) for quantification of tacrolimus and active metabolites after extraction from whole blood is described. The lower limit of detection was 2 microg/L. Intraassay precision (CV) was 5.7-13.7%, and the interassay CV was 6. 1-14.9%. Tacrolimus trough concentrations in 104 whole blo...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1998-12-01 00:00:00
abstract：:We developed a new colorimetric method, DNA enzyme immunoassay (DEIA), for detecting specific hybrids of complementary nucleic acids and applied it to the detection of hepatitis B virus (HBV) DNA amplified from serum samples by means of the polymerase chain reaction (PCR) technique. The method is based on the ability ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1991-03-01 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：:Using Syva "EMIT" reagents and the Cobas Bio centrifugal analyzer, we have developed cost-effective assays for therapeutic drug monitoring of carbamazepine, phenobarbital, and phenytoin. With these optimized methods one can assay up to 2470 samples with a single 100-test EMIT kit with good precision and accuracy. Betw...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1989-08-01 00:00:00
abstract：:Four routine assays commonly used for monitoring plasma methotrexate (MTX) during high-dose therapy were validated by HPLC as the comparison method. MTX and its main metabolite, 7-hydroxymethotrexate (7-OHMTX), were analyzed by HPLC with postcolumn derivatization and fluorometric detection. About 200 clinical plasma s...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1996-01-01 00:00:00
abstract：:A highly sensitive two-site enzyme immunoassay (Capcellia) was developed to determine the concentration of CD4 and CD8 molecules expressed on the surface of human T lymphocytes. This assay, performed in one step (20 min), involves the specific immunocapture of T lymphocytes and reaction of the CD4 or CD8 molecules wit...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1994-01-01 00:00:00
abstract：BACKGROUND:Pompe disease (PD) is the first lysosomal storage disorder to be added to the Recommended Uniform Screening Panel for newborn screening. This condition has a broad phenotypic spectrum, ranging from an infantile form (IOPD), with severe morbidity and mortality in infancy, to a late-onset form (LOPD) with vari...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2016.259036
更新日期：2017-04-01 00:00:00
abstract：BACKGROUND:In vitro and animal studies suggest that tumor necrosis factor alpha (TNF-alpha) modulates intestinal iron transport. We hypothesized that the effect of TNF-alpha might be particularly relevant if iron absorption is not effectively controlled by the HFE gene. METHODS:In patients with homozygous C282Y hemoch...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2005.065417
更新日期：2006-08-01 00:00:00
abstract：:We describe a modification of the MacGee method [Anal. Chem. 42, 421 (1970)] for rapid determination of phenytoin (diphenylhydantoin) in plasma by gas-liquid chromatography, in which an ethylaing reagent is used instead of the more common methylating reagents. With this modification, several N-methylated antiepileptic...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1975-05-01 00:00:00
