Abstract：
BACKGROUND:New genetic forms of hereditary hemochromatosis (HH) or hereditary hyperferritinemia (HF) have been identified over the last few years, and abnormalities of various genes may interact in a single patient. This study aimed to develop a rapid automated method for sequencing the main genes involved. METHODS:We used a standard 96-well microplate with a single PCR condition in an adaptation of the SCAIP (single-condition amplification with internal primer) method to sequence the HFE (hemochromatosis), HAMP (hepcidin antimicrobial peptide), HFE2/HJV [hemochromatosis type 2 (juvenile)], SLC40A1 (ferroportin), and TFR2 (transferrin receptor 2) genes, and the 5' untranslated region of the FTL (ferritin, light polypeptide) gene. To further simplify the method, we adjusted PCR conditions to avoid the use of an internal primer and applied this single-condition amplification method to 38 selected, unrelated patients. We tailored the genetic investigation according to the clinical picture, with the patients falling into 2 groups. Group 1 consisted of patients with hyperferritinemia and high transferrin saturation (TS) (classic adult and juvenile HH forms, groups 1A and 1B, respectively), and group 2 consisted of patients with hyperferritinemia and low, typical, or slightly increased TS, with or without iron overload (groups 2A and 2B, respectively). RESULTS:With this strategy we identified single-gene and multigene abnormalities, including 6 previously undescribed abnormalities in HFE (c.794dupA), HFE2 (c.-89-4dupT), and SLC40A1 (c.262A>G, c.533G>A, c.1468G>A, and c.-59_-45del). CONCLUSION:This method is a simple approach for investigating hereditary iron overload or HF and allows rapid evaluation of patients.
journal_name
Clin Chemjournal_title
Clinical chemistryauthors
Cunat S,Giansily-Blaizot M,Bismuth M,Blanc F,Dereure O,Larrey D,Quellec AL,Pouderoux P,Rose C,Raingeard I,Renard E,Schved JF,Aguilar-Martinez P,CHU Montpellier AOI 2004 Working Group.doi
10.1373/clinchem.2007.090605subject
Has Abstractpub_date
2007-12-01 00:00:00pages
2060-9issue
12eissn
0009-9147issn
1530-8561pii
clinchem.2007.090605journal_volume
53pub_type
杂志文章abstract：:We compared a stabilized-temperature L'vov platform furnace containing an end-heated graphite atomizer (HGA) and transverse Zeeman background-correction system with a side-heated furnace system (transversely heated graphite atomizer; THGA) containing a longitudinal Zeeman background-correction system for the determina...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1994-03-01 00:00:00
abstract：:We determined the LD-1/LD-2 isoenzyme ratio in hemolysates of erythrocytes by electrophoresis on cellulose acetate and on agarose. A ratio exceeding 1.0 was found with the former but not the latter. Results were similar for in vitro models of hemolytic disorders. Using cellulose acetate electrophoresis, we determined ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1981-08-01 00:00:00
abstract：BACKGROUND:C-reactive protein (CRP), the prototypic marker of inflammation, is present in atherosclerotic plaques and appears to promote atherogenesis. Also, CRP has been localized to monocytes and tissue macrophages, which are present in the necrotic core of lesions prone to plaque rupture. Leukocyte-derived myelopero...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2008.109207
更新日期：2009-02-01 00:00:00
abstract：BACKGROUND:High-resolution DNA melting analysis of small amplicons is a simple and inexpensive technique for genotyping. Microfluidics allows precise and rapid control of temperature during melting. METHODS:Using a microfluidic platform for serial PCR and melting analysis, 4 targets containing single nucleotide varian...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2017.276147
更新日期：2017-10-01 00:00:00
abstract：:A radioimmunoassay for barbiturates is shown to be equally sensitive to phenobarbital and its major urinary metabolite, p-hydroxyphenobarbital, in serum and urine. Interference by the metabolite can be essentially eliminated by selectively extracting phenobarbital into chloroform. The extraction efficiency of the meth...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1977-05-01 00:00:00
abstract：BACKGROUND:Measurements of the ionic current flowing through nanometer-scale pores (nanopores) have been used to analyze single DNA and RNA molecules, with the ultimate goal of achieving ultrafast DNA sequencing. However, attempts at purely electronic measurements have not achieved the signal contrast required for sing...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2007.091231
更新日期：2007-11-01 00:00:00
abstract：:We studied the effects on 25 analytes of duration of contact of serum with non-anticoagulated blood and of temperature. Serum was separated after blood was allowed to stand, for 0, 2, 4, 6, 8, 24, or 48 h at 4, 23, or 30 degrees C. Results obtained for bilirubin, albumin, zinc sulfate turbidity, thymol turbidity, chol...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1981-01-01 00:00:00
abstract：:Time-resolved fluoroimmunoassay was developed for the detection of synovial-type phospholipase A2 (s-PLA2) in human serum. This solid-phase, sandwich assay uses a polyclonal rabbit antibody raised against synovial-type group II PLA2 produced in Escherichia coli. No cross-reactions were detected between s-PLA2 and PLA2...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1992-09-01 00:00:00
abstract：:Diabetic patients in poor glycemic control show increased glycation of total plasma proteins, but little is yet known about the relative extents to which the various individual proteins are glycated. Thus, we studied the non-enzymic glycation of several major plasma proteins and plasma protein fractions in normal and ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1987-12-01 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：BACKGROUND:The genotype of hepatitis C virus (HCV) is a predictor of antiviral therapeutic response. We describe an approach for HCV genotype determination by real-time PCR and melting curve analysis. METHODS:After automated nucleic acid extraction, we used reverse transcription-PCR in a block cycler to amplify nucleo...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：2002-12-01 00:00:00
abstract：:We report a case of Fanconi's anemia with an extremely high proportion of fetal hemoglobin (Hb F). A three-year-old girl with multiple birth defects, mental retardation, and aplastic anemia consistent with Fanconi's anemia showed Hb AF by electrophoresis; the Kleihauer smear showed Hb F in 70% of her erythrocytes. Tot...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1985-10-01 00:00:00
abstract：:We report the accuracy, imprecision, total analytical errors, and patient misclassification errors for cholesterol measured from capillary whole blood, venous whole blood, and venous plasma samples by five devices used in public cholesterol screening environments: Reflotron, Vision, Ektachem DT-60, QuickRead, and Lipo...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1993-02-01 00:00:00
abstract：:We describe a liquid-chromatographic procedure for simultaneously determining eight steroids in serum. We used a Zorbax ODS column and a mobile phase of methanol/isopropanol/water (44/10/46, by vol), which well resolves the steroids cortisone, cortisol, corticosterone, 11-deoxycortisol, 11-deoxycorticosterone, androst...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1987-08-01 00:00:00
abstract：BACKGROUND:Thrombopoietin (TPO), the major hormone controlling platelet production, has been measured in thrombocytopenias with discordant results. The aim of our work was to assess the value of the TPO assay for differential diagnosis of thrombocytopenias in a large cohort of patients classified according to the resul...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：2001-09-01 00:00:00
abstract：BACKGROUND:Group B streptococcal infections are the leading cause of sepsis and meningitis in newborns. A rapid and reliable method for the detection of this pathogen at the time of delivery is needed for the early treatment of neonates. Isothermal amplification techniques such as recombinase polymerase amplification h...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2013.213504
更新日期：2014-04-01 00:00:00
abstract：:We have developed an interactive statistical quality-control system for the small- to medium-sized radioimmunoassay laboratory, which can be used in a programmable desk-top calculator instead of the medium- or large-scale computer systems usually required. The design of this quality-control system is modeled after the...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1978-07-01 00:00:00
abstract：BACKGROUND:Concomitant quantification of multiple mutant KRAS (v-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog) alleles may provide information in addition to that provided by standard mutation-detection procedures. We assessed the feasibility of a nanofluidic digital PCR array platform to detect and quantify KRAS...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2012.186577
更新日期：2012-09-01 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：:The currently available radioimmunoassays of trypsin measure total immunoreactive trypsin (EC 3.1.1.7), which includes both trypsinogen and alpha 1-protease inhibitor-bound trypsin. Hitherto, the only way to differentiate these two forms of trypsin has been to fractionate them on a gel-filtration column. We describe h...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1986-09-01 00:00:00
abstract：:We developed and evaluated a direct ultraviolet method for the enzymatic determination of uric acid in serum, plasma, or urine, without deproteinization of sera and plasma. Equilibrium and nonlinear curve-fitting kinetic options are evaluated and compared, and results of the proposed method are compared with those of ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1986-03-01 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：:I describe a simple, single-tube batch fractionation procedure for separating MM and MB isoenzymes of creatine kinase on a macroporous strong anion exchanger (AG MP-1, Bio-Rad Laboratories). The isoenzymes can be separated in less than 3 min, with a resulting dilution of the serum with no more than an equal volume of ...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1976-01-01 00:00:00
abstract：BACKGROUND:We have developed a silicon-based biosensor that generates a visual signal in response to nucleic acid targets. METHODS:In this system, capture oligonucleotide probes are immobilized on the surface of the biosensor. Interaction of the capture probes with a complementary target and a biotinylated detector ol...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1999-09-01 00:00:00
abstract：:The formal definition of sensitivity associates the term with the change in the response of a system for a small change of the stimulus causing the response, i.e., the ratio of the response of a system to the stimulus causing it. One interpretation of sensitivity associates the rate of change of the response for a sma...
journal_title：Clinical chemistry
pub_type： 评论,杂志文章
doi：
更新日期：1997-10-01 00:00:00
abstract：:Over the past decade, significant improvements in immunochemical and electrophoretic techniques have enabled collection of heretofore unavailable data on proteins in biological fluids, greatly increasing our understanding of protein physiology in the various body compartments and providing the foundation for clinical ...
journal_title：Clinical chemistry
pub_type： 杂志文章,评审
doi：
更新日期：1982-05-01 00:00:00
abstract：:The apolipoprotein A-I (apo A-I) radioimmunoassay established in our laboratory involves use of purified apo A-I as the primary standard for quantifying apo A-I in plasma and a pooled plasma (quality-control sample) as a secondary standard to decrease interassay variation. The measured values for apo A-I increased ove...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1986-07-01 00:00:00
abstract：:The use of 4-chlorotestosterone acetate by farmers for cattle fattening was recently demonstrated although the use of this anabolic steroid is strictly forbidden in the European Union. We investigated the metabolism of 4-chlorotestosterone acetate in the bovine species after intramuscular and oral administration. Nine...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1998-05-01 00:00:00
abstract：:In this enzyme-linked immunoassay for human thyrotropin (TSH) in unextracted serum we use 96-well immunoenzymometric assay plates, first coated with polyclonal antibody to TSH, then incubated with the serum samples and reacted with mouse monoclonal antibody to human TSH. After incubation with alkaline phosphatase-labe...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1985-07-01 00:00:00
abstract：:We adapted an inhibition assay for methotrexate, involving dihydrofolate reductase from bovine liver, for use with a discrete analyzer (the ABA-100). The analyzer was used both for dilution and a 5-min pre-incubation of the sample with NADPH--enzyme reagent, and for the assay itself. The standard curve was linear betw...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1980-02-01 00:00:00
