Abstract:
:We developed a new colorimetric method, DNA enzyme immunoassay (DEIA), for detecting specific hybrids of complementary nucleic acids and applied it to the detection of hepatitis B virus (HBV) DNA amplified from serum samples by means of the polymerase chain reaction (PCR) technique. The method is based on the ability of an anti-DNA monoclonal antibody to discriminate between single-stranded and double-stranded DNA. A solid phase was coated with a specific oligonucleotide probe, internal to the amplified region of HBV DNA, via an avidin-biotin bridge. The denatured PCR product was hybridized with the solid-phase probe, and the amplified DNA probe hybrid was then incubated with a monoclonal antibody specific for double- but not single-stranded DNA. Colorimetric detection of the DNA-antibody complex was achieved by adding an anti-mouse Ig antibody labeled with horseradish peroxidase. The combined use of DEIA and PCR can reveal a few HBV genome copies present in a serum sample. This method has several advantages: (a) the sensitivity is adequate for the detection of amplified DNA; (b) the signal is associated with the hybridization event, independently of modifications of the probe or of the amplification primers; and (c) the test is simple and rapid and, most importantly, requires only the standard facilities of a routine clinical laboratory.
journal_name
Clin Chemjournal_title
Clinical chemistryauthors
Mantero G,Zonaro A,Albertini A,Bertolo P,Primi Dsubject
Has Abstractpub_date
1991-03-01 00:00:00pages
422-9issue
3eissn
0009-9147issn
1530-8561journal_volume
37pub_type
杂志文章abstract:BACKGROUND:The management of critically ill infants and neonates includes frequent determination of arterial blood gas, electrolyte, and hematocrit values. An objective of attached point-of-care patient monitoring is to provide clinically relevant data without the adverse consequences associated with serial phlebotomy....
journal_title:Clinical chemistry
pub_type: 杂志文章,多中心研究
doi:
更新日期:2002-11-01 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::Four double-monoclonal time-resolved immunofluorometric assays (TrIFMAs) have been developed for the specific determination of pregnancy-associated plasma protein A/proeosinophil major basic protein (PAPP-A/ proMBP) complex in first-trimester maternal serum samples. The assays have a functional sensitivity of < 4 mIU/...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1997-12-01 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::Quantification of ethanol by a radiative energy attenuation (REA) technique was evaluated and compared with results by gas-liquid chromatography (GLC) and by the Du Pont aca. Within-assay CVs were less than 5.5%. Between-assay CVs ranged from 1.9% to 6.0% for serum and blood controls at concentrations of 0.5, 1.0, and...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1984-11-01 00:00:00
abstract::We determined fluoxetine (Prozac) and its major metabolite norfluoxetine in plasma by liquid chromatography with fluorescence detection. After liquid-liquid extraction from 1 mL of plasma, the extract was derivatized at room temperature with dansyl chloride, and the highly fluorescent derivatives were chromatographed ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1992-09-01 00:00:00
abstract:BACKGROUND:Although cardiac troponin (cTn) is a cornerstone marker in the assessment and management of patients with acute coronary syndrome (ACS) and heart failure (HF), cTn is not diagnostically specific for any single myocardial disease process. This narrative review discusses increases in cTn that result from acute...
journal_title:Clinical chemistry
pub_type: 杂志文章,评审
doi:10.1373/clinchem.2009.130799
更新日期:2009-12-01 00:00:00
abstract::We report the adaptation of four radioassays to the prototype of an automated radioimmunoassay system ("Centria," Union Carbide). The system consists of three integrated modules: (a) an automated pipettor, which dispenses samples and reagents; (b) the key module, an incubator/separator, in which centrifugal force is u...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1975-08-01 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::A modified fermentation procedure based on one described by Homer Smith in his textbook was used to eliminate glucose interference in the automated detrmination of inulin. The manifold of Fjeldbo and Stamey [J. Lab. Clin. Med. 72, 353 (1968)] was modified to enable us to insert the fermentation procedure without sacri...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1977-07-01 00:00:00
abstract:BACKGROUND:Recent studies have suggested that glial fibrillary acidic protein (GFAP) serum concentrations distinguish between intracerebral hemorrhage (ICH) and ischemic stroke (IS) shortly after symptom onset. In this prospective multicenter trial we validated GFAP in an independent patient cohort and assessed the qua...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2016.263335
更新日期:2017-01-01 00:00:00
abstract:BACKGROUND:Since its beginnings, newborn screening for cystic fibrosis (CF) using an assay for immunoreactive trypsinogen (IRT) has been plagued by a high rate of false-positive results (screen positive, diagnosis negative), despite attempts to reduce this rate by use of altered cutoffs and second-tier DNA testing. IRT...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2009.132480
更新日期:2010-03-01 00:00:00
abstract:BACKGROUND:Hyperhomocysteinemia (HHcy) has been linked to impaired left ventricular function and clinical class in patients with chronic heart failure. We hypothesized that HHcy stimulates myocardial brain natriuretic peptide (BNP) expression and induces adverse left ventricular remodeling. METHODS:We randomized 50 ra...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2006.077859
更新日期:2007-04-01 00:00:00
abstract:BACKGROUND:High circulating concentrations of N-terminal fragments of A- and B-type natriuretic peptides (NT-proANP and NT-proBNP) identify patients with impaired cardiac function. ProANP-derived peptides are particularly sensitive to increased preload of the heart and proBNP-derived peptides to increased afterload; th...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2004.039891
更新日期:2005-04-01 00:00:00
abstract::Taurine was measured in cerebrospinal fluid by reacting it with fluorescamine to form a fluorescent derivative, followed by separation on a reversed-phase column and fluorometric detection and evaluation. The assay is rapid (17 min) and sensitive to as little as 1 mumol/L. The mean value for 27 cerebrospinal fluid sam...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1979-08-01 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:
更新日期:1989-03-01 00:00:00
abstract::Measurement of the mass concentration of serum enzymes by radioimmunoassay provides direct quantitation of specific isoenzymes and may be less subject to some of the limitations of traditional assay procedures for enzymes. We describe the development of a sensitive and specific radioimmunoassay for the muscle isoenzym...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1978-03-01 00:00:00
abstract::A highly sensitive two-site enzyme immunoassay (Capcellia) was developed to determine the concentration of CD4 and CD8 molecules expressed on the surface of human T lymphocytes. This assay, performed in one step (20 min), involves the specific immunocapture of T lymphocytes and reaction of the CD4 or CD8 molecules wit...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1994-01-01 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
abstract::At the clinical level, we have seen the emergence of a subspecialty of affective disorders within psychiatry. Mood clinics, depression units, and affective disorders centers are appearing in many academic and clinical settings, where clinical skill and knowledge can be concentrated and new research furthered. By using...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1988-05-01 00:00:00
abstract:BACKGROUND:Recent publications have explored the possibility of using fingerprints to confirm drug use, but none has yet dealt with environmental contamination from fingertips. Here we explored the possibility of establishing an environmental cutoff for drug testing from a single fingerprint. METHODS:Fingerprint sampl...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2017.281469
更新日期:2018-06-01 00:00:00
abstract:BACKGROUND:The multiple endocrine neoplasia 2 (MEN 2) syndromes [MEN 2A, MEN 2B, and familial medullary thyroid carcinoma (FMTC)] are caused by germline mutations of the RET protooncogene. Because 85% of MEN 2A patients and 30% of FMTC patients have mutations at codon 634, the recommended molecular analyses begin at ex...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:2001-11-01 00:00:00
abstract::Market forces have dramatically influenced the environment in which healthcare is delivered, but these changes do not need to be interpreted negatively by community laboratorians. Only total vertical integration of laboratory medicine can control episode-of-care cost. Opportunities also exist for horizontal integratio...
journal_title:Clinical chemistry
pub_type: 杂志文章,评审
doi:
更新日期:1996-05-01 00:00:00
abstract::Erythrocyte protoporphyrin content is increased in several human and veterinary disorders in which heme synthesis is disrupted. The nature of the defect determines whether zinc protoporphyrin, unchelated (free) protoporphyrin, or both are present in the erythrocytes. Extraction of the protoporphyrin into acetone/water...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1981-02-01 00:00:00
abstract:BACKGROUND:Microsatellite instability (MSI) is a useful phenotype in cancer diagnosis and prognosis. Nevertheless, methods to detect MSI status from next generation DNA sequencing (NGS) data are underdeveloped. METHODS:We developed an approach to detect the MSI phenotype using NGS (mSINGS). The method was used to eval...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2014.223677
更新日期:2014-09-01 00:00:00
abstract::We measured the cross reactivity of two medications--digitoxin and spironolactone--in two digoxin radioimmunoassay (liquid and solid-phase) kit procedures. Both tests showed similar average percentages of cross reactivity with digitoxin (7.2 and 8.9% for intravenous, and 11.9 and 10.9% for oral administration), but no...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1978-04-01 00:00:00
abstract::The Dutch Foundation for Quality Assessment in Clinical Chemistry (SKZL) is the professional organization that conducts external quality-assessment schemes in The Netherlands. However, such schemes in fact assess the performance of the internal quality-control systems of the participating laboratories. In this paper w...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1991-07-01 00:00:00
abstract::We describe a novel ion-selective electrode (ISE) system that can be used in the Abbott Vision analyzer. A reusable sensor pack and a disposable test pack have been miniaturized to approximately the size and weight of a conventional Vision test pack, thus eliminating the need for a separate ISE module. The sensor pack...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1990-12-01 00:00:00
abstract:BACKGROUND:Subfractions of HDL, particularly large HDL (HDL2), are inversely correlated with the severity of coronary artery disease (CAD). alpha-Tocopherol (AT) is the main lipid-soluble antioxidant in plasma. Results of a previous small study (n = 44) suggested that either a combination of an antioxidant cocktail [80...
journal_title:Clinical chemistry
pub_type: 杂志文章,随机对照试验
doi:10.1373/clinchem.2006.078865
更新日期:2007-03-01 00:00:00
abstract::We describe a mechanized method for centrifugal analyzer determination of sorbitol dehydrogenase in serum, based on conversion of D-fructose to sorbitol with simultaneous oxidation of NADH, in triethanolamine buffer at pH 7.4 and 30 degrees C. The standard curve for this assay is linear to 200 U of activity per liter ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1979-12-01 00:00:00