Abstract:
BACKGROUND:A single nonfunctional chimeric gene with its 5' and 3' ends corresponding to CYP21P and CYP21, respectively, is caused by unequal gene crossover in the CYP21 genes during meiosis. The presence of the chimeric CYP21P/CYP21 molecule can not be detected by conventional PCR methods and therefore may be lost in PCR amplification. This leads to a false result and diagnostic discordance. METHODS:We developed a rapid and direct method to detect a chimeric CYP21P/CYP21 gene that uses a 3'-specific primer for the CYP21 gene and two different 5' primers for both CYP21 and CYP21P to amplify the wild-type CYP21 and the chimeric CYP21P/CYP21 genes. A secondary PCR that can differentiate the chimeric from the wild-type gene was also performed. The PCR product was directly analyzed on agarose gel. RESULTS:After careful titration, we found that earlier failure to detect the chimeric CYP21P/CYP21 gene could be caused by unequal concentrations of two independent alleles as the PCR template or by the lack of primers to amplify chimeric molecules. We successfully amplified the chimeric gene using our improved method. CONCLUSIONS:The chimeric CYP21P/CYP21 is present in a large portion of congenital adrenal hyperplasia patients. By adding a CYP21P/CYP21-specific primer, we were able to amplify and detect both homozygous and heterozygous chimeric genes. Therefore, our new PCR-based assay is a more effective way to analyze congenital adrenal hyperplasia mutations.
journal_name
Clin Chemjournal_title
Clinical chemistryauthors
Lee HH,Chang JG,Tsai CH,Tsai FJ,Chao HT,Chung Bsubject
Has Abstractpub_date
2000-05-01 00:00:00pages
606-11issue
5eissn
0009-9147issn
1530-8561journal_volume
46pub_type
杂志文章abstract::Sensitivity, stability, and specificity of the color-producing reaction of hydrogen peroxide with bezidine, leuco-malachite green, or o-dianisidine were tested in numerous systems containing hemoglobin and other hemoproteins. Use of urea or low temperature (to minus 12 degrees C), or both, was highly beneficial, espec...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1975-03-01 00:00:00
abstract::In this simple and low-cost radioimmunoassay (RIA) procedure for thyroxin (T4), ethacridine (6,9-diamino-2-ethoxyacridine lactate; Rivanol) is used to separate antibody-bound ligand and free ligand. This acridine dye precipitates free T4 not bound to antibody. At pH 8.6, it precipitates more than 90% of 125I-labeled T...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1983-11-01 00:00:00
abstract::We describe a simple and convenient method for processing small amounts of tissue samples for trace-metal measurements by atomic absorption spectrometry, by use of a modified Parr microwave digestion bomb. Digestion proceeds rapidly (less than or equal to 90 s) in a sealed Teflon-lined vessel that eliminates contamina...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
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journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2006.083311
更新日期:2007-04-01 00:00:00
abstract::The purpose of Medicare's Demonstration of Competitive Bidding for Clinical Laboratory Services is to determine whether competitive bidding can be used to provide quality laboratory services at prices below current Medicare reimbursement rates. Here, we present key features of the preliminary design for the demonstrat...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1998-08-01 00:00:00
abstract:BACKGROUND:Monogenic autoinflammatory diseases are caused by pathogenic variants in genes that regulate innate immune responses, and are characterized by sterile systemic inflammatory episodes. Since symptoms can overlap within this rapidly expanding disease category, accurate genetic diagnosis is of the utmost importa...
journal_title:Clinical chemistry
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doi:10.1093/clinchem/hvaa024
更新日期:2020-04-01 00:00:00
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journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1985-04-01 00:00:00
abstract::The reaction of 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB, Ellman's reagent) with thiols is sensitive to daylight, in particular to ultraviolet radiation at wavelengths around 325 nm. Exposure to light at the absorbance maximum of the yellow product (the thionitrobenzoate ion) at 410 nm had no effect on the reaction. ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1987-10-01 00:00:00
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journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2016.256768
更新日期:2016-07-01 00:00:00
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journal_title:Clinical chemistry
pub_type: 杂志文章,评审
doi:10.1373/clinchem.2010.156901
更新日期:2011-02-01 00:00:00
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journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2007.091488
更新日期:2007-12-01 00:00:00
abstract::A highly sensitive sandwich-type chemiluminescence enzyme immunoassay for plasma endothelin-I (ET-1), involving no extraction steps, has been developed. Two populations of polyclonal antibodies were used in the present study: One is specific to the C-terminus of the endothelin family of peptides; the other, a Fab' fra...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1996-08-01 00:00:00
abstract::Determination of the concentration of erythrocyte folate by means of competitive protein binding assay critically depends on the extraction procedure applied. Results will be influenced by variable factors such as the in vitro age of the blood samples, the degree of hemolysis, the presence of ascorbic acid, and the pH...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1976-07-01 00:00:00
abstract::Antiserum performance in a nephelometric system can be characterized by parameters derived from measuring reaction rates. The characterization process is derived from a series of dose-response curves (elicited nephelometric response vs antigen concentration) generated from various dilutions of the antiserum being test...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1981-11-01 00:00:00
abstract::To correct the results of a radioimmunoassay for beta-thyrotropin (TSH) subunit by eliminating the proportion ascribable to intact TSH, we have devised a method that experimentally reproduces the conditions under which the interference develops. Beta-TSH subunit was assayed in several preparations containing known con...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1987-05-01 00:00:00
abstract::We describe the development of a stable reference material for prostatic acid phosphatase, derived from human prostatic tissue and human seminal fluid. The enzyme was purified by an L-tartramic acid affinity-chromatography technique. Two-dimensional electrophoresis revealed essentially no contaminating proteins, and s...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1984-08-01 00:00:00
abstract::We describe a procedure for determining thiocyanate in plasma with a Technicon AutoAnalyzer I, modified with a 50-mm tubular flow cell to increase the sensitivity of thiocyanate detection. The procedure is suitable for distinguishing cigarette smokers from non-smokers, the mean for smokers being 151 (SD 43) micromol/L...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1980-03-01 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::We investigated the efficiency, accuracy, and reliability of the ultrafiltration/dipstick methodology commonly used to diagnose myoglobinuria. Twenty-five myoglobin-containing urine specimens were filtered by centrifugation for 15 min at 1500g through a Centricon-30 membrane filter. Both the original specimen and filt...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1996-06-01 00:00:00
abstract::In considering factors that might influence measurement of free thyroxin (T4), we evaluated the proportion (%) of T4 that could be stripped from thyroxin-binding globulin (TBG). The percentage of free T4 was measured in serially diluted sera from four normal subjects, four patients with hyperthyroidism or hypothyroidi...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1990-02-01 00:00:00
abstract::Glucose was measured by the ferricyanide, the Beckman glucose oxidase, and the hexokinase procedures in 228 plasma samples taken during standard oral glucose-tolerance tests in 17 normal subjects and in 21 chemical diabetics. The neocuproine method was also used to measure glucose concentration in 156 samples (78 befo...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1982-12-01 00:00:00
abstract::I describe an assay for serum estriol during pregnancy, which does not require prior extraction or chromatography of the serum. Fifty microliters of 20-fold diluted serum can be assayed to detect as little as 20 pg of estriol. For a 20 mug/liter serum estriol concentration the CV was 16%; for 100 mug/liter concentrati...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1976-04-01 00:00:00
abstract:BACKGROUND:Direct equilibrium dialysis and direct ultrafiltration free thyroxine (T(4)) assays rely on semipermeable membranes to exclude T(4)-binding serum proteins from dialysates and ultrafiltrates. The presence of these proteins in dialysates or ultrafiltrates will yield spuriously high free T(4) values when free T...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2007.085316
更新日期:2007-05-01 00:00:00
abstract::We measured thyroid hormones and thyroxin-binding proteins in serum from 62 normal lactating and 52 nonlactating women at three months postpartum, and compared these values with those for 42 nonpregnant control women of similar age. Mean thyroxin concentrations in the lactating and nonlactating women were significantl...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1987-07-01 00:00:00
abstract:BACKGROUND:Atherosclerosis is a systemic disease with focal rupture of vulnerable plaque responsible for major clinical events. Several population-based studies indicate an association between lipoprotein-associated phospholipase A2 (Lp-PLA2) and cardiovascular events. Lp-PLA2 is emerging as a biomarker that may be a p...
journal_title:Clinical chemistry
pub_type: 杂志文章,评审
doi:10.1373/clinchem.2006.070672
更新日期:2006-09-01 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::Establishing associations between environmental agents and disease presents challenges to both epidemiologists and toxicologists, particularly in cases of complex gene-environment interactions and when there is a long latency between exposure and disease. Biologic markers, physiological signals that reflect exposure, ...
journal_title:Clinical chemistry
pub_type: 杂志文章,评审
doi:
更新日期:1994-07-01 00:00:00
abstract:BACKGROUND:In this study we demonstrate the technical application of flow cytometry and cell sorting combined with gene-rearrangement clonality profiling to detect and confirm minimal disease in 2 leukemia and 2 lymphoma cases. METHODS:Specimens with low percentages (0.05%-5%) of abnormal lymphoid populations were ide...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2005.061259
更新日期:2006-03-01 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