Abstract：
BACKGROUND:A unique requirement for the molecular diagnosis of mitochondrial DNA (mtDNA) disorders is the ability to detect heteroplasmic mtDNA mutations and to distinguish them from homoplasmic sequence variations before further testing (e.g., sequencing) is performed. We evaluated the potential utility of temporal temperature gradient gel electrophoresis (TTGE) for these purposes in patients with suspected mtDNA mutations. METHODS:DNA samples were selected from patients with known mtDNA mutations and patients suspected of mtDNA disorders without detectable mutations by routine analysis. Six regions of mtDNA were PCR amplified and analyzed by TTGE. Electrophoresis was carried out at 145 V with a constant temperature increment of 1.2 degrees C/h. Mutations were identified by direct sequencing of the PCR products and confirmed by PCR/allele-specific oligonucleotide or PCR/restriction fragment length polymorphism analysis. RESULTS:In the experiments using patient samples containing various amounts of mutant mtDNA, TTGE detected as little as 4% mutant heteroplasmy and identified heteroplasmy in the presence of a homoplasmic polymorphism. In 109 specimens with 15 different known mutations, TTGE detected the presence of all mutations and distinguished heteroplasmic mutations from homoplasmic polymorphisms. When 11% of the mtDNA genome was analyzed by TTGE in 104 patients with clinically suspected mitochondrial disorders, 7 cases of heteroplasmy ( approximately 7%) were detected. CONCLUSIONS:TTGE distinguishes heteroplasmic mutation from homoplasmic polymorphisms and appears to be a sensitive tool for detection of sequence variations and heteroplasmy in patients suspected of having mtDNA disorders.
journal_name
Clin Chemjournal_title
Clinical chemistryauthors
Chen TJ,Boles RG,Wong LJsubject
Has Abstractpub_date
1999-08-01 00:00:00pages
1162-7issue
8 Pt 1eissn
0009-9147issn
1530-8561journal_volume
45pub_type
临床试验,杂志文章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：:The measurement of glycohemoglobin (GHb) by boronate affinity chromatography is useful in monitoring long-term glucose control in diabetic subjects. The inherent disadvantage of this method is the hands-on time required because the hemoglobin fractions are separated on individual disposable columns. To overcome this d...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1993-01-01 00:00:00
abstract：BACKGROUND:SELDI-TOF mass spectrometry (MS) is a high-throughput proteomic approach with potential for identifying novel forms of serum biomarkers of arthritis. METHODS:We used SELDI-TOF MS to analyze serum samples from patients with various forms of inflammatory arthritis. Several protein profiles were collected on d...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2007.099549
更新日期：2008-06-01 00:00:00
abstract：:Plasma concentrations of delta-9-tetrahydrocannabinol, the principal psychoactive cannabinoid in marijuana, decline to values substantially less than 1 microgram/L within a few hours after a subject has smoked a marijuana cigarette. Using a single-quadrupole gas chromatograph-mass spectrometer (GC/MS) operated in the ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1991-12-01 00:00:00
abstract：BACKGROUND:Genomic instability resulting in copy number variation is a hallmark of malignant transformation and may be identified through massive parallel sequencing. Tumor-specific cell free DNA (cfDNA) present in serum and plasma provides a real-time, easily accessible surrogate. METHODS:DNA was extracted from serum...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2014.226571
更新日期：2015-01-01 00:00:00
abstract：BACKGROUND:Recent studies have suggested that single-stranded DNA (ssDNA) library preparation can enrich short DNA species from the plasma of healthy individuals, cancer patients, and transplant recipients. Based on previous observations that fetal DNA molecules in the maternal plasma are shorter than maternal DNA mole...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2016.268656
更新日期：2017-05-01 00:00:00
abstract：:We have developed and validated simple, rapid immunoassays to measure concentrations of albumin in urine ranging from 5 to 200 mg/L. We use antiserum to human albumin, raised in sheep, and we separate the antibody-bound and free fractions of albumin by using a second antiserum, produced in donkeys, against the Fc frag...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1986-07-01 00:00:00
abstract：BACKGROUND:Cystic fibrosis is a life-threatening genetic disorder that has been associated with mutations in the CFTR [cystic fibrosis transmembrane conductance regulator (ATP-binding cassette sub-family C, member 7)] gene. Hundreds of CFTR mutations have been detected to date. Current CFTR genotyping assays target a s...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2013.206466
更新日期：2013-10-01 00:00:00
abstract：:We report here the normal range of serum alkaline phophatase activity as measured by the method proposed by Hausamen et al. [Clin. Chim. Acta 15, 241 (1967)] with a much larger sample size than used in previous investigations. In the statistical analysis the sample population is subdivided by sex and age, two variable...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1977-09-01 00:00:00
abstract：:We developed a radioimmunoassay with a gamma-emitting radioligand to measure androstenedione in human serum and in dried blood-spot samples from newborns. Antisera were raised in rabbits against androstenedione linked to bovine serum albumin at positions 3, 6, or 11 on the steroid nucleus. Radioligands were prepared b...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1989-08-01 00:00:00
abstract：BACKGROUND:The susceptibility of plasma lipids to oxidation is thought to be a factor contributing to atherogenic risk. Various groups have studied the in vitro oxidizability of isolated LDL and examined the effects of conventional antioxidants. The drawbacks associated with the isolation of LDL for evaluation of in vi...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2005.051078
更新日期：2005-11-01 00:00:00
abstract：:We evaluated the performance of two homogeneous assays for quantifying HDL cholesterol (HDL-C) and compared them with the phosphotungstic acid (PTA)/ MgCl2 assay. Both homogeneous HDL-C assays were precise, having a within-run CV of < 1.20% and a between-run CV of < 4.07%. The HDL-C values (y) measured by the two homo...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1997-06-01 00:00:00
abstract：BACKGROUND:Adiponectin is an adipocyte-derived hormone circulating in different multimer complexes. The high-molecular-weight (HMW) complex is likely the active form of this protein and has been recognized as a risk marker for type 2 diabetes and coronary artery disease (CAD). Because quantification of HMW adiponectin ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2008.112425
更新日期：2009-03-01 00:00:00
abstract：:This simple polarization fluoroimmunoassay for detection of amphetamine in urine involves the use of a single reagent: suitably diluted antiserum plus fluorescein-labeled amphetamine as tracer. To this, one adds 10 microL of urine, incubates for a few minutes at room temperature, and measures the fluorescence polariza...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1985-07-01 00:00:00
abstract：:The clinical utilities of the emerging biochemical markers for mammary adenocarcinoma CA 15-3, CA 549, CA M26, CA M29, and MCA (mucin-like carcinoma-associated antigen) were assessed by a formal rating according to six desirable marker characteristics. All five indicators similarly have good specificities but limited ...
journal_title：Clinical chemistry
pub_type： 杂志文章,评审
doi：
更新日期：1993-11-01 00:00:00
abstract：INTRODUCTION:Parathyroid hormone-related peptide (PTHrP) is involved in activating pathways, allowing tumor cells to form bone metastases. Measurement of PTHrP is used for the diagnosis and clinical management of patients suspected of hypercalcemia of malignancy. We developed an LC-MS/MS method for measuring PTHrP, est...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2015.244012
更新日期：2016-01-01 00:00:00
abstract：BACKGROUND:Age-adjusted morbidity and mortality rates from coronary heart disease (CHD) are higher in men than in women. Androgens are suspected to be responsible for the male disadvantage. The genomic effect of androgens is mediated by the androgen receptor (AR), which has a polymorphic CAG repeat in exon 1. The numbe...
journal_title：Clinical chemistry
pub_type： 杂志文章,多中心研究
doi：10.1373/clinchem.2005.049262
更新日期：2005-07-01 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：:We measured total serum bile acids on a fluorescence-light-scattering micro centrifugal analyzer by the direct enzymatic method with 3 alpha-hydroxysteroid dehydrogenase (EC 1.1.1.50) and with resazurin as a fluorogenic electron acceptor. We found that serum protein has an inhibitory effect on the measurement of bile ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1983-01-01 00:00:00
abstract：:A simple computer-based screening technique has been developed for classifying human expired air components into 16 chemical classes, based on empirical formulas. The sort procedure was developed to simplify the screening of the composition of expired air samples by sorting all components into chemical classes and cla...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1988-08-01 00:00:00
abstract：:An oligonucleotide "melting" procedure was developed whereby we can monitor for a sequence heterogeneity in the gene for apolipoprotein E production. Two oligonucleotides were synthesized, one recognizing the common epsilon 3 allele but not the common epsilon 2 allele, the other recognizing the epsilon 2 allele but no...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1986-07-01 00:00:00
abstract：BACKGROUND:The fibroblast growth factor (FGF) and FGF receptor (FGFR) axis plays a critical role in tumorigenesis, but little is known of its influence in ovarian cancer. We sought to determine the association of genetic variants in the FGF pathway with risk, therapeutic response, and survival of patients with ovarian ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2013.211490
更新日期：2014-01-01 00:00:00
abstract：BACKGROUND:Standardization of hemoglobin (Hb)A(1c) measurements is a process of considerable interest for quality assurance in diabetes management. To contribute to continuous quality improvement and fulfillment of the requirements for reference measurement procedures according to the standards of the International Org...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2009.139477
更新日期：2010-05-01 00:00:00
abstract：:A new method (enzyme-ligand immunoassay, ELIA) is described for the estimation of estrogen (ER) and progesterone (PR) receptors in microsamples of human breast cancer tissue. The technique, based on the nonisotopic measurement of receptor-bound estradiol and progesterone, involves three steps: (a) simultaneous saturat...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1995-01-01 00:00:00
abstract：BACKGROUND:Cardiovascular mortality is markedly higher in Russia than in Western Europe and the US. Little is known about indicators of atherosclerotic risk in the Russian population. To our knowledge, this is the first study of apolipoprotein (apo) AI and B in Russia based on the WHO-IFCC standard. METHODS:We measure...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2003.023853
更新日期：2004-02-01 00:00:00
abstract：:We have developed an automated latex agglutination test for Treponema pallidum (TPLA) for measurement of the concentration of treponemal antibodies in syphilitic sera. The assay needs neither a complicated pretreatment of the sera nor special techniques. Intra- and interassay precision studies showed high reproducibil...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1993-08-01 00:00:00
abstract：BACKGROUND:The hepatocyte-derived acute-phase reactant C-reactive protein (CRP) has been the subject of intense research over the last 2 decades for its possible role in the pathogenesis of cardiovascular diseases. This research has spawned interest in the use of the blood concentration of CRP for predicting a first co...
journal_title：Clinical chemistry
pub_type： 杂志文章,评审
doi：10.1373/clinchem.2008.115923
更新日期：2009-02-01 00:00:00
abstract：:The objective of this study was to develop a clinical laboratory method for subclass typing of human immunoglobulin G (IgG) paraproteins. Serum proteins were isoelectrically focused (IEF) in a mini-gel and passively blotted by capillary diffusion onto untreated nitrocellulose. Unreacted sites on the nitrocellulose wer...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1989-03-01 00:00:00
abstract：BACKGROUND:Lectin-like oxidized LDL receptor 1 (LOX-1) is implicated in atherothrombotic diseases. Activation of LOX-1 in humans can be evaluated by use of the LOX index, obtained by multiplying the circulating concentration of LOX-1 ligands containing apolipoprotein B (LAB) times that of the soluble form of LOX-1 (sLO...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2009.140707
更新日期：2010-04-01 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
