Abstract：
BACKGROUND:In the workup of patients with suspected hereditary nonpolyposis colorectal cancer (HNPCC), detection of loss of heterozygosity (LOH) could help pinpoint the mismatch-repair (MMR) gene carrying the germline mutation, but analysis of microsatellite markers has proved unreliable for this purpose. We developed a simple, low-cost method based on single-nucleotide polymorphism (SNP) genotyping and capillary electrophoresis for the assessment of LOH at 2 MMR loci simultaneously. METHODS:We used the Applied Biosystems SNaPshot Multiplex Kit with meticulously selected primers to assess 14 common SNPs in MLH1 [mutL homolog 1, colon cancer, nonpolyposis type 2 (E. coli)] and MSH2 [mutS homolog 2, colon cancer, nonpolyposis type 1 (E. coli)] and optimized the protocol for DNA isolated from peripheral blood and fresh/frozen or archival microsatellite-unstable tumors from patients with confirmed (n = 42) or suspected (n = 25) HNPCC. The 42 tumors from patients with confirmed MLH1 or MSH2 germline mutations were used to validate the method's diagnostic accuracy against results obtained with DNA sequencing or multiplex ligation-dependent probe amplification. RESULTS:The SNaPshot assay provided better detection of certain SNPs than DNA sequencing. The MLH1 and MSH2 SNP marker sets were informative in 82% and 76% of the 67 cases analyzed, respectively. The new assay displayed 100% specificity for detecting LOH and predicted the location of the germline mutation in 40% of the cases (54% of those involving MLH1, 22% in MSH2). CONCLUSIONS:Our SNP-based method for detecting LOH in MLH1 and MSH2 is simple to perform with instruments available in most clinical genetics laboratories. It can be a valuable addition to protocols now used to guide mutational screening of patients with suspected HNPCC.
journal_name
Clin Chemjournal_title
Clinical chemistryauthors
Bujalkova M,Zavodna K,Krivulcik T,Ilencikova D,Wolf B,Kovac M,Karner-Hanusch J,Heinimann K,Marra G,Jiricny J,Bartosova Zdoi
10.1373/clinchem.2008.108902subject
Has Abstractpub_date
2008-11-01 00:00:00pages
1844-54issue
11eissn
0009-9147issn
1530-8561pii
clinchem.2008.108902journal_volume
54pub_type
杂志文章abstract：:We used flameless atomic absorption spectrophotometry to measure concentrations of Fe, Mo, Li, As, and Zn in cerebrospinal fluid (CSF) of patients with malignant brain tumors benign brain tumors, non-brain malignant tumors and control (non-neoplastic disease) patients. Mean (and SD) concentrations (microgram/L) of the...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1986-12-01 00:00:00
abstract：:We have modified an automated measurement system of urinary iodine (UI) and established a sensitive UI assay system by using ultraviolet (UV) digestion. The automated system is sensitive enough to detect concentrations of UI < 0.78 mumol/L (< 10 micrograms/dL) in a small volume of urine (500 microL). Sample throughput...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1995-04-01 00:00:00
abstract：:We measured immunoglobulins in the sera of 33 patients on days 1, 3, 6, 10, and 17 and three to four weeks after surgical operations (mostly hysterectomy or appendectomy) or (six patients) after spinal injury. In the absence of infection or blood transfusion, IgG usually decreased slightly and transiently after hyster...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1975-05-01 00:00:00
abstract：BACKGROUND:The single cell represents the basic unit of all organisms. Most investigations have been performed on large cell populations, but understanding cell dynamics and heterogeneity requires single-cell analysis. Current methods for single-cell analysis generally can detect only one class of analytes. METHODS:Re...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2012.191445
更新日期：2012-12-01 00:00:00
abstract：:The concentrations of 25 plasma proteins were measured in 22 patients with membranous nephropathy. For some large proteins, the plasma concentrations were increased; there were also large proteins with low plasma concentrations, but small or medium-sized proteins showed uniformly lower plasma concentration than the co...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1997-07-01 00:00:00
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 precision of estimates of the sensitivity of diagnostic tests is evaluated. "Sensitivity" is defined as the fraction of diseased subjects with test values exceeding the 0.975-fractile of the distribution of control values. An estimate of the sensitivity is subject to sample variation because of variation of both c...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1985-04-01 00:00:00
abstract：:We developed a simple method for the analysis of bismuth (Bi) in biological tissue, using wet digestion for sample pretreatment. Bi was determined by electrothermal atomic absorption spectrometry with platinum as a matrix modifier to decrease the volatility of Bi. The furnace program included a gas stop for sensitivit...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1993-05-01 00:00:00
abstract：:We describe a simple polarization fluoroimmunoassay for the detection of several commonly abused drugs in urine. The single reagent used, prepared by mixing four different antisera and three fluorescein-labeled derivatives, produces a polarization signal that is the average of the individual signals of the derivatives...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1987-10-01 00:00:00
abstract：:We have separated serum proteins in only 15 min by high-performance liquid chromatography. The identification of several peaks in the chromatographic profile was greatly aided by the use of multiple-wavelength detection. We have found a good correlation between retention times and electrophoretic mobilities. Serum sam...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1980-09-01 00:00:00
abstract：BACKGROUND:Although lipoprotein-associated phospholipase A(2) (Lp-PLA(2)) levels are associated with cardiovascular events, Lp-PLA(2) is physically linked to LDL cholesterol (LDL-C). Whether measures of Lp-PLA(2) mass or activity continue to predict risk after LDL-C reduction by statin therapy is uncertain. METHODS:Lp...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2011.180281
更新日期：2012-05-01 00:00:00
abstract：:We describe a new and specific method for measurement of lecithin palmitic acid in amniotic fluid. Dipalmitoyl lecithin, the major alveolar surfactant, has previously been estimated by measuring the lecithin-sphingomyelin ratio, total lecithin, total phospholipid phosphorus, and (or) total palmitic acid. Our method is...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1977-01-01 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
abstract：:Inherited qualitative abnormalities of fibrinogen have been documented in more than 100 families. These dysfibrinogenemias usually are clinically silent, but in some cases are associated with bleeding, thrombosis, or defective wound healing. Abnormalities of the fibrinogen molecule may impair any of the major steps in...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1985-04-01 00:00:00
abstract：:Clinically relevant interassay precision profiles for thyrotropin (thyroid-stimulating hormone; TSH) were constructed with human serum pools measured over 4-8 weeks by six immunometric assays, in at least two different reagent lots. Functional sensitivities (the concentration at which the interassay CV is < or = 20%) ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1995-03-01 00:00:00
abstract：:Even though significant progress has been made in the development of electrodes for the in vivo detection of catecholamines, oxygen, and various cations, there has been little research on the feasibility of continuously monitoring drugs in whole blood by electrochemical sensing devices. Electroanalytical problems asso...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1982-09-01 00:00:00
abstract：:Each of 65 laboratories analyzed 10 whole-blood samples for erythrocyte protoporphyrin by one or more of several analytical procedures. These procedures were of two types: (a) extraction of protoporphyrin from the erythrocytes into ethyl acetate/acetic acid, re-extraction into hydrochloric acid, and fluorometric measu...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1978-12-01 00:00:00
abstract：BACKGROUND:Measurement of purine and pyrimidine metabolites presents complex problems for separations currently performed by HPLC and thin-layer chromatography in clinical practice. We developed a novel capillary electrophoresis method for this purpose. METHODS:Separations were performed in 60 mmol/L borate-2-amino-2-...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1999-12-01 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：:We describe an enzyme-linked immunosorbent assay for determination of light-chain ratios for IgG, IgA, and IgM in serum. A commercial serum with known overall kappa and lambda concentrations was used as standard. To capture the respective immunoglobulins, we used antibodies to gamma, lambda and mu chains coated onto m...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1990-03-01 00:00:00
abstract：BACKGROUND:D-2-Hydroxyglutaric aciduria (D-2-HGA), L-2-hydroxyglutaric aciduria (L-2-HGA), and the combined D/L-2-hydroxyglutaric aciduria (D/L-2-HGA) are poorly understood organic acidurias. To investigate the usefulness of cultured human skin fibroblasts for both diagnostic and research purposes, we measured disease-...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/49.7.1133
更新日期：2003-07-01 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：BACKGROUND:The analytical and clinical performance of homogeneous LDL-cholesterol assays has been reported, but their reactions with subfractions of LDL and VLDL have not been described in detail. METHODS:We evaluated reaction selectivity of two homogeneous LDL-cholesterol assays, LDLk (Kyowa Medex) and LDLd (Daiichi ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：2002-11-01 00:00:00
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-c...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1997-08-01 00:00:00
abstract：:Zinc protoporphyrin is the predominent fluorescent porphyrin accumulating in erythrocytes as a result of chronic lead absorption or iron-deficiency anemia. Although diagnostic concentrations or thresholds for it in erythrocytes have been inferred, normal adult averages have not been established. We quantitatively asse...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1980-09-01 00:00:00
abstract：:A method is presented for the assay of delta-aminolaevulinate dehydratase (EC 4.2.1.24) to be performed on 10 microL of whole blood. Results compare favorably with those obtained with an accepted method (J. Clin. Chem. Clin. Bio-chem. 12: 389-390, 1974) that involves use of 200 microL of blood. ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1981-01-01 00:00:00
abstract：:We describe a multiprobe colorimeter that permits the reading of enzyme-linked immunosorbent assays (ELISA) in situ, by use of microhemagglutination plates. The use of dipping probes eliminates interference from meniscus effects and air-bubble entrapment. Detailed validation tests have been done and potential sources ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1979-04-01 00:00:00
abstract：BACKGROUND:Iron may promote coronary atherosclerotic disease (CAD) by increasing lipid peroxidation. Studies on biochemical or genetic markers of body iron stores as risk factors for CAD have yielded conflicting results. METHODS:We studied 849 individuals with a clear-cut definition of the CAD phenotype, i.e., with (C...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：2002-01-01 00:00:00
abstract：BACKGROUND:Plasma B-type cardiac natriuretic peptides reflect cardiac structure and function and have proven roles in assisting in the diagnosis of acute heart failure. They are independent prognostic indicators across the full spectrum of cardiovascular disease. Serial changes in plasma B-type cardiac natriuretic pept...
journal_title：Clinical chemistry
pub_type： 杂志文章,评审
doi：10.1373/clinchem.2011.165704
更新日期：2012-01-01 00:00:00
abstract：:The IFCC-recommended procedure for aspartate aminotransferase (EC 2.6.1.1) was adapted to the centrifugal analyzer and evaluated during five years. The main hindrance to widespread use of the recommended method is the need for pre-incubation with pyridoxal phosphate. The present method includes a 10-min pre-incubation...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1981-01-01 00:00:00
