Abstract:
BACKGROUND:Suspension array technology has surpassed ELISA for automated, simultaneous detection and quantification of soluble biomarkers such as virus-specific antibodies. We describe assays in which antigens are attached to a lipid bilayer surrounding color-coded particles. METHODS:We used layer-by-layer technology to establish a multiplex suspension array with distinguishable microbeads coated with authentic viral surfaces to catch and quantify virus-specific antibodies in a flow cytometric analysis. Antigenic surfaces were generated by chimeric and wild-type baculoviruses plus 2 different influenza A virus subtypes fused to a lipid bilayer surrounding distinctly colored particles. Specificity of binding of chosen antibodies and sera was detected by immunofluorescence. Results of multiplex analysis were compared with results of ELISA. RESULTS:Titrations of virus-specific antibodies in the multiplex suspension array demonstrated specific binding to the viral surface proteins. The multiplex suspension array gave positive results for up to log 5-diluted primary antibodies with an approximately 5- to 10-fold reduced dynamic range compared with the respective ELISA. CONCLUSIONS:The bead-based multiplex suspension array is customizable and easy to establish. By displaying native influenza A virus surfaces and recombinant HIV-1 epitopes, the new assay provides a tool for the detection of major viral infections in humans.
journal_name
Clin Chemjournal_title
Clinical chemistryauthors
Toellner L,Fischlechner M,Ferko B,Grabherr RM,Donath Edoi
10.1373/clinchem.2005.065789subject
Has Abstractpub_date
2006-08-01 00:00:00pages
1575-83issue
8eissn
0009-9147issn
1530-8561pii
clinchem.2005.065789journal_volume
52pub_type
杂志文章abstract::The manual Reference Method of the Centers for Disease Control for serum iron (CDC/RM/Fe) and a semiautomated adaptation of it were used to evaluate two working methods: one, a detergent solubilization procedure for the Roche Cobas-Bio analyzer, the other, the Kodak Ektachem 700 procedure, based on dry-film technology...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1990-10-01 00:00:00
abstract:BACKGROUND:The TDx Fetal Lung Maturity II (FLM II) assay uses amniotic fluid to assess lung maturity of the unborn infant. We investigated common preanalytical factors that influence FLM results, including centrifugation, sample storage, and contamination by whole blood. METHODS:We tested 18 specimens after centrifuga...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/49.6.935
更新日期:2003-06-01 00:00:00
abstract:BACKGROUND:Increased concentrations of cell-free DNA have been found in several disorders and have been interpreted as evidence of increased rates of cell death or turnover. Evidence from in vitro and animal experiments suggests that DNA may play a role in the pathogenesis of rheumatoid arthritis (RA). METHODS:We meas...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2006.084509
更新日期:2007-09-01 00:00:00
abstract::We describe a method used for determining serotonin and tryptamine in urine after oral loading with their precursor amino acid. Several factors affecting prepurification and chromatographic separation of both indoleamines were optimized, including sample extraction and mobile-phase composition. Under optimal condition...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1989-01-01 00:00:00
abstract::Highly purified human lactate dehydrogenase 1 has been used in an interlaboratory evaluation and improvement progran in clinical chemistry in New York State since 1971. Although there are difficulties in determining and assigning the most nearly accurate values for test samples in the absence of a reference method and...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1977-06-01 00:00:00
abstract::We describe a modified tantalum ribbon atomic absorption procedure for determining lead in undiluted whole blood. An instrumentation Laboratory (I.L.) Model 151 atomic absorption spectrophotometer equipped with an I.L. Model 355 Flameless Sampler was used. The Flameless Sampler was slightly modified to include three-c...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1978-07-01 00:00:00
abstract::For normal and above-normal concentrations of angiotensin-converting enzyme (ACE; EC 3.4.15.1) activity in plasma, results of a manual fluorometric method [with hippuryl-histidyl-leucine (HHL), 5 mmol/L, as substrate] correlated well with those of an automated spectrophotometric method [with 3-(2-furylacryloyl)-L-phen...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1991-08-01 00:00:00
abstract::We describe an enzyme-linked immunosorbent assay for determination of light-chain ratios for IgG, IgA, and IgM in serum. A commercial serum with known overall kappa and lambda concentrations was used as standard. To capture the respective immunoglobulins, we used antibodies to gamma, lambda and mu chains coated onto m...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1990-03-01 00:00:00
abstract::We investigated two techniques of immunoblotting--the Western blot and the dot blot--for use in detecting prostatic acid phosphatase (PAP, EC 3.1.3.2). We used polyclonal antisera to human PAP, produced in rabbits by hyperimmunization with purified PAP, and PAP-specific monoclonal antibodies in the immunoenzymatic pro...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1986-10-01 00:00:00
abstract::Detection and characterization of small IgM paraproteins by immunoelectrophoresis are often difficult because of (a) slow diffusion of the macromolecular immunoglobulin, and (b) the obscuring of the light chain, especially kappa, by the light chains of background polyclonal immunoglobulin (the "umbrella effect"). Vari...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1988-08-01 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::This new assay procedure for diaphorase eliminates problems of high blank rates and nonlinear kinetics associated with other methods. The dye thiazolyl blue tetrazolium bromide is reduced in the presence of NADH and diaphorase to yield a colored formazan, which as maximum absorbance at 560 nm. ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1979-12-01 00:00:00
abstract::In this liquid-chromatographic method for the determination of bromide in human serum, an important and unique feature is pre-analysis ultrafiltration of the sample, which effectively removes proteins. The chromatographic system consists of an anion-exchange column (Whatman Partisil SAX 10-micron particles), an isocra...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1984-05-01 00:00:00
abstract::In the Prospective Cardiovascular Münster (PROCAM) study, serum lipoprotein(a) [Lp(a)] and its relationship to pro- and anticoagulatory as well as fibrinolytic indices were determined in a large group of employees: 864 men (m) and 373 women (f), ages 16-65 years. Univariate statistical analysis showed Lp(a) concentrat...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1991-11-01 00:00:00
abstract::The fructosamine test is considered clinically useful for assessing short-term integrated control of blood glucose, but there are few published data to support this hypothesis. We fractionated glycated and nonglycated proteins by affinity chromatography on phenylboronate columns and, with specific immunochemical metho...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1987-10-01 00:00:00
abstract::Numerous papers have been written to show which combinations of Shewhart-type quality-control charts are optimal for detecting systematic shifts in the mean response of a process, increases in the random error of a process, and linear drift effects in the mean response across the assay batch. One paper by Westgard et ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1993-08-01 00:00:00
abstract::An enzymatic, fluorometric method is described for determination of serum urea on silicone-rubber pads. In this method, the reagents are lyophilized on the surface of the pads, NADH on one side and a mixture of urease, glutamate dehydrogenase, and alpha-ketoglutarate on the other. The rate of disappearance of NADH flu...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1975-01-01 00:00:00
abstract::Gene probes can now be used to detect a variety of mutations that produce single-gene disorders. In present clinical practice, restriction endonuclease analysis is used for the prenatal diagnosis of sickle cell anemia, alpha-thalassemia, and beta-thalassemia. Direct detection of the mutation is possible in alpha-thala...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1985-09-01 00:00:00
abstract:BACKGROUND:Nandrolone is one of the most abused anabolic steroids, and its use in doping is increasing, as revealed by numerous positive cases during recent years in various sports. Different authors have reported the possible natural production of nandrolone metabolites in humans, and some of these authors argued that...
journal_title:Clinical chemistry
pub_type: 临床试验,杂志文章
doi:10.1373/clinchem.2003.022848
更新日期:2004-02-01 00:00:00
abstract::The method is based on the selective activating capacity of dithiothreitol on creatine kinase isoenzyme MB, after isoenzyme MM is activated by glutathione. Isolated isoenzymes MM and MB of human and canine origin were assayed individually and in mixtures of known activities. When glutathione was present in the assay m...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1975-10-01 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::We present data on a filter-paper collection and assay method for measurement of glycated hemoglobin (gHb). Onto filter paper dipped into a solution of glucose oxidase (EC 1.1.3.4) and allowed to dry, approximately 20 microL of capillary blood was spotted. For analysis, we eluted the dried blood spot from the paper by...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1986-05-01 00:00:00
abstract::Purified arylsulfatase A (EC 3.1.6.1) from human urine was radioiodinated under conditions that caused no significant loss of antigenic activity. We used this labeled arylsulfatase A (specific radioactivity 4-7.5 Ci/g) together with nonlabeled enzyme and rabbit antiserum produced against homogeneous enzyme to develop ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1985-03-01 00:00:00
abstract:BACKGROUND:Improved methods are required to screen drug candidates for their influences on protein interactions. There is also a compelling need for miniaturization of screening assays, with attendant reductions in reagent consumption and assay costs. METHODS:We used sensitive, miniaturized proximity ligation assays (...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2007.099424
更新日期:2008-07-01 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::A fixed-time incubation method for measuring acetylsalicylic acid esterase was improved by using a higher concentration of the substrate, acetylsalicylic acid, and an activator, Ca2+. The enzymatic activity in serum was 25-fold that measured by earlier methods. Inhibition studies showed a pattern similar to that repor...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1983-03-01 00:00:00
abstract::We describe use of near-infrared reflectance spectroscopy (NIRRS) to estimate the amount of fat in feces, for diagnosis of steatorrhea. After sample homogenization, the spectrum of the fecal homogenate is scanned over the near-infrared region. Assay of 94 samples of feces having a known concentration of fat showed the...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1987-04-01 00:00:00
abstract:BACKGROUND:B-type natriuretic peptide (BNP), which is activated in heart failure (HF), is processed to an active form by corin. The corin gene is expressed in the human heart and kidney, but corin protein expression in the heart, kidney, and circulation, along with whether proBNP is processed by circulating corin, rema...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2010.153908
更新日期:2011-01-01 00:00:00
abstract::A new method for the reliable identification of localized variations in DNA by detection of associated diagnostic products with matrix-assisted laser desorption ionization time-of-flight mass spectrometry is described. The diagnostic products are generated by the primer oligo base extension (PROBE) reaction, which req...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1997-07-01 00:00:00
abstract::We evaluated the EMIT (enzyme-multiplied immuno technique) and FPIA (fluorescence polarization immunoassay) urine screening systems for detection of benzodiazepine intake. Healthy male volunteers were given single oral therapeutic doses of alprazolam (2 mg), chlordiazepoxide (25 mg), flunitrazepam (1 mg), lorazepam (3...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1992-02-01 00:00:00