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 facilitate rapid screening of clinically relevant TPMT mutations, we developed a strategy of high-throughput genotyping by applying denaturing HPLC (DHPLC). METHODS:To test the specificity and efficiency of the DHPLC method, 98 DNA samples from a selected population of patients receiving thiopurine therapy or with previous thiopurine withdrawal were analyzed for the most frequent mutant TPMT alleles, *2 and *3A, which contain key mutations in exons 5, 7, and 10 to identify clearly different elution profiles. All fragments were examined by direct sequencing. Additionally, to test the sensitivity of DHPLC analysis, genotyping for the *2 and *3A alleles of all 98 DNA samples was performed by PCR-based methods (PCR-restriction fragment polymorphism analysis and allele-specific PCR). RESULTS:The presence of mutations discriminating for alleles *2, *3A, *3C, and *3D, as well as various silent and intron mutations, were correctly predicted by DHPLC in 100% of the samples as confirmed by direct sequencing. Comparison with PCR-based methods for alleles *2 and *3 produced an agreement of 100% with no false-negative signals. CONCLUSIONS:DHPLC offers a highly sensitive, rapid, and efficient method for genotyping of the relevant TPMT mutations, discriminating at least for alleles *2 and *3, in clinical and laboratory practice. Additionally, DHPLC allows a simultaneous screening for novel genetic variability in the TPMT gene.
journal_name
Clin Chemjournal_title
Clinical chemistryauthors
Schaeffeler E,Lang T,Zanger UM,Eichelbaum M,Schwab Msubject
Has Abstractpub_date
2001-03-01 00:00:00pages
548-55issue
3eissn
0009-9147issn
1530-8561journal_volume
47pub_type
杂志文章abstract：:We evaluated a new agarose-gel-electrophoretic procedure (Corning) (I) for separating and quantitating of high-density lipoprotein cholesterol (HDLC), comparing it with the modified Lipid Research Clinics (LRC) procedure (heparin 183 kilounits/L, MnCl2 92 mmol/L) (II). Method I was insensitive to an HDLC concentration...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1979-11-01 00:00:00
abstract：:We describe a mechanized chromogenic assay for factor X, the results of which correlate well with those for the one-stage clotting assays for factor X in which it is activated either via the extrinsic pathway by thromboplastin or directly by Russell's viper venom. We purified human factor X and raised monospecific ant...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1980-06-01 00:00:00
abstract：:We compare measurement of total fast hemoglobin (HbA1) by "high-performance" liquid chromatography and by electrophoresis on agar gel. Blood samples were obtained from a diverse population (n = 222): offspring of two diabetic parents, diabetic patients with and without retinopathy, diabetic and non-diabetic pregnant w...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1981-03-01 00:00:00
abstract：:Red blood cell (RBC) replacement solutions are being developed as alternatives to allogeneic RBC use in blood transfusions in the treatment of massive trauma, to achieve hemodynamic stability during elective surgery, and to increase oxygen-carrying capacity in anemia. Hemoglobin-based oxygen carrier (HBOC)-201 (Biopur...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1997-09-01 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：BACKGROUND:Reduced renal function is an established risk factor for cardiovascular events. We compared 3 measures of renal function--serum cystatin C, serum creatinine, and calculated creatinine clearance--as predictors of subsequent cardiovascular events in a community-based population of elderly individuals. METHODS...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2009.135962
更新日期：2010-05-01 00:00:00
abstract：:Using image-analyzing equipment, we measured the effect of quinidine on the activity of naphthol AS-D chloroacetate esterase, alpha-naphthyl butyrate esterase, alpha-naphthyl acetate esterase, and acid phosphatase in individual cells of a human Hep G2 hepatoma cell line. The impact of the drug on the morphology of the...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1988-03-01 00:00:00
abstract：:The commonly used methods of assessing the precision of the automated leukocyte differential have certain drawbacks that affect the validity and comparability of results. In the present report, we introduce a procedure based on building precision profiles from a large number of within-run imprecision experiments. The ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1996-07-01 00:00:00
abstract：:Using a solid-phase monoclonal antibody enzyme immuno-assay, we evaluated in a multicenter study (18 laboratories) the utility of evaluating catalytic activity of human placental alkaline phosphatase (hPLAP, EC 3.1.3.1) in serum as a potential tumor marker. We determined hPLAP in serum samples from 130 patients with o...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1988-10-01 00:00:00
abstract：:In the present clinical evaluation of Triage immunoassay for detection of drugs of abuse, we examined a possible connection between the laboratory skills of the personnel using it and the quality of analytical results; we also evaluated the degree of concordance between the Triage results and those obtained by an inst...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1996-09-01 00:00:00
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journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1977-09-01 00:00:00
abstract：:Six alpha-amylase (EC 3.2.1.1) isoenzymes have been resolved electrophoretically on cellulose acetate membranes in a discontinuous buffer system. The fastest migrating isoenzymes are of salivary origin (S1, S2, S3), the slower ones of pancreatic origin (P1, P2, P3). We determined the amylase isoenzyme distribution in ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1976-01-01 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：:Major depression is one of the most common clinical problems seen by primary-care physicians. The prevalence is 5% to 10% in primary-care outpatients and 15% in medically ill inpatients. Despite this high prevalence rate, depression is frequently not accurately diagnosed. Evidence suggests that misdiagnosis often occu...
journal_title：Clinical chemistry
pub_type： 杂志文章,评审
doi：
更新日期：1988-05-01 00:00:00
abstract：BACKGROUND:Increased plasma troponin T (cTnT), but not troponin I (cTnI), is frequently observed in end-stage renal failure patients. Although generally considered spurious, we previously reported an associated increased mortality at 12 months. METHODS:We studied long-term outcomes in 244 patients on chronic hemodialy...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：2001-03-01 00:00:00
abstract：BACKGROUND:Data regarding the prognostic value of programmed cell death ligand 1 (PD-L1) expression on circulating tumor cells (CTCs) are lacking. However, CTCs could represent an alternative approach to serial biopsies, allowing real-time monitoring of cancer phenotype. METHODS:We evaluated, in a dedicated prospectiv...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1093/clinchem/hvaa121
更新日期：2020-08-01 00:00:00
abstract：:We report an easy, rapid method for quantifying bone isoenzyme of alkaline phosphatase (EC 3.1.3.1., ALP) in serum. The original method described by Rosalki and Ying Foo (Clin Chem 1984;30:1182-6) was somewhat simplified. In contrast to their results, we found that bone ALP is precipitated quantitatively by wheat-germ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1986-10-01 00:00:00
abstract：BACKGROUND:Recent recognition of its broad pathophysiological importance has triggered an increased interest in 25-hydroxyvitamin D [25(OH)D]. By consequence, throughput in 25(OH)D testing has become an issue for clinical laboratories, and several automated assays for measurement of 25(OH)D are now available. The aim o...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2011.176545
更新日期：2012-03-01 00:00:00
abstract：:In this rapid, cost-effective, enzyme-linked immunosorbent assay for 17 alpha-hydroxyprogesterone (17-OHP) eluted from dried blood spotted on filter paper, second antibody is coated onto the microwell plate and horseradish peroxidase (EC 1.11.1.7) is the label enzyme. Antiserum to 17-OHP was prepared by using 4-(2-car...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1987-06-01 00:00:00
abstract：BACKGROUND:Emerging technologies for rapid identification of microbes demonstrate a shift from traditional biochemical and molecular testing algorithms toward methods using mass spectrometry (MS) for the semiquantitative analysis of microbial proteins and genetic elements. This study was performed to assess the diagnos...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2011.161968
更新日期：2011-07-01 00:00:00
abstract：BACKGROUND:Increased cardiac troponin concentrations in acute coronary syndrome (ACS) identify patients with ongoing cardiomyocyte necrosis who are at increased risk. However, with the use of more precise assays, cardiac troponin increases are commonly noted in other cardiovascular conditions as well. This has generate...
journal_title：Clinical chemistry
pub_type： 杂志文章,评审
doi：10.1373/clinchem.2016.261495
更新日期：2017-01-01 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：PURPOSE:To review information on the use of laboratory tests in screening, diagnosis, and monitoring of acute and chronic hepatic injury. DATA SOURCES AND STUDY SELECTION:A MEDLINE search was performed for key words related to hepatic diseases, including acute hepatitis, chronic hepatitis, alcoholic hepatitis, cirrhos...
journal_title：Clinical chemistry
pub_type： 杂志文章,评审
doi：
更新日期：2000-12-01 00:00:00
abstract：BACKGROUND:The Dutch project "Calibration 2000" aims at harmonization of laboratory results via calibration by development of matrix-based secondary reference materials. We considered the selection, preparation, and characterization of 34 potential reference materials (PRMs). METHODS:Sixteen PRMs were prepared either ...
journal_title：Clinical chemistry
pub_type： 杂志文章,多中心研究
doi：
更新日期：2002-09-01 00:00:00
abstract：BACKGROUND:We previously reported a family in which the propositus and both her father and paternal grandmother had high serum aminopeptidase N (CD13; EC 3.4.11.2) activity (autosomal dominant). The molecular mass of the serum CD13 polypeptide of the propositus was larger than that of normal CD13, suggesting either a m...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：2001-02-01 00:00:00
abstract：BACKGROUND:Band 5 tartrate-resistant acid phosphatase (TrACP; EC 3. 1.3.2) consists of two isoenzymes, bands 5a and 5b, of which band 5b TrACP is considered to be derived from bone. However, no kinetic method for the specific measurement of band 5b TrACP in serum is available. Our aim was to develop a kinetic assay met...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：2000-04-01 00:00:00
abstract：:This case demonstrates the importance of monitoring theophylline concentrations in serum during therapy with this drug. The commonly used body-weight-derived guidelines for therapeutic dose are not adequate and do not preclude the necessity for such monitoring. The ultraviolet spectrophotometric methods available for ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1978-09-01 00:00:00
abstract：:Development of a new, sensitive immunoassay for measuring transforming growth factor beta 1 (TGF-beta1) is described and compared with four commercially available TGF-beta1 immunoassays. Preanalytical conditions were evaluated. The nonlinearity found in serum or plasma is due to masking of TGF-beta1 by binding protein...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1997-10-01 00:00:00
abstract：:In this automated system for assaying guanidino compounds, the compounds are resolved on an ion-exchange (Dowex 50) column, then reacted with phenanthrenequinone in alkaline solution. This mixture is then acidified and the resulting fluorescence is measured (lambda ex, 305 nm; lambda em, 395 nm). I applied the system ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1984-02-01 00:00:00
abstract：:We compared the ionic activity of sodium, as measured with glass electrodes, with sodium concentration in 23 healthy persons, 15 persons with acute renal failure, and before and after dialysis in 46 patients with chronic renal failure. In healthy persons the mean (+/- SEM) sodium concentration was 139.1 +/- 0.6 mmol/L...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1985-11-01 00:00:00
