Development of a Genotype 325–Specific proCPU/TAFI ELISA (original) (raw)
Related papers
Measurement of aminoterminal propeptide of type I procollagen (PINP) in serum
Clinical Biochemistry, 2012
Background: Recently, measurement of amino terminal propeptide of type III procollagen (PIIINP) was introduced as a part of the hepatic cirrhotic marker enhanced liver fibrosis TM test on the automated ADVIA Centaur ® immunoassay platform (Siemens Healthcare Diagnostics Inc., Tarrytown, NY, USA). In this article, we show that the Centaur PIIINP may be used in place of the much more labor-intensive RIA method, and we present an age stratified reference interval. Methods: We analyzed four control samples 20 times over a period of 5 days. Centaur PIIINP assay measurements were compared with the widely used UniQ PIIINP RIA assay (Orion Diagnostica, Espoo, Finland) using 55 patient samples (range = 3.7-43.3 µg/L). Furthermore, we established a reference interval based on samples from 287 blood donors. Results: In the concentration range 2.5-11.9 µg/L, the total imprecision was below 8%. Comparison with the UniQ PIIINP RIA assay yielded: Centaur PIIINP µg/L = 1.9 × (UniQ PIIINP RIA)+0.6 µg/L, r 2 = 0.94. The reference interval for the Centaur PIIINP assay showed no gender difference but was stratified by age [4.0-11.0 µg/L (18-40 years) and 3.5-9.5 µg/L (41-70 years)]. Conclusions: We conclude that the Centaur PIIINP assay is suitable for routine use with our newly defined reference interval. The results obtained by Centaur correlates well with those obtained by the previously employed RIA, though the absolute values are higher.
Enzyme immunoassay using a rat prolactin-alkaline phosphatase recombinant tracer
Analytical Chemistry, 1993
This paper describes a competitive enzyme immunoassay of rat prolactin (rPrl) using a recombinant conjugate as a colorimetric tracer. rPrl was inserted into the N-terminal end of Escherichia coli alkaline phosphatase (AP), using a n expression vector which allows insertion of foreign DNA sequences between codons +6 and +7 of the phoA gene. The assay was performed in 96-well microtiter plates coated with a mouse monoclonal antibody raised against rabbit immunoglobulin G.
Comparison of immunoturbidimetric and immunonephelometric assays for specific proteins
Clinical Biochemistry, 2009
Immunoturbidimetric assays for specific proteins are available on "open system" clinical chemistry analyzers. The analytical performance of nine immunoturbidimetric specific protein assays (C3, C4, CRP, Haptoglobin, IgA, IgG, IgM, RF, and Transferrin) was compared to immunonephelometry. Testing was performed on the Abbott ARCHITECT ci8200 and the Dade Behring BNII nephelometer and evaluated for precision, linearity, limit of detection, prozone phenomenon, method comparison, workflow, and proficiency testing survey comparison. Immunoturbidimetric assays performance was satisfactory for total precision, linearity, limit of detection and the prozone effect was not observed. Method comparison was acceptable for the immunoglobulins, CRP and transferrin but less favorable for the other assays, likely due to methodology and antibody specificity differences. Immunourbidimetric specific protein assays allow for efficient test consolidation on a general purpose clinical chemistry analyzer.
Molecular and Genetic Analyses of the Putative Proteus O Antigen Gene Locus
Applied and Environmental Microbiology, 2010
databases. Finally, Proteus-specific genes from these five serogroups were identified by screening 79 strains belonging to the 68 Proteus O antigen serogroups. To our knowledge, this is the first molecular characterization of the putative Proteus O antigen locus, and we describe a novel molecular classification method for the identification of different Proteus serogroups.
Pathology, 1999
This study compares the concordance of results in different ELISAs for antineutrophil cytoplasmic antibodies (ANCA) directed against proteinase 3 (PR3) or myeloperoxidase (MPO). Sera were considered ªtrue positivesº if they were positive according to the manufacturer's criteria in a least three of the five PR3-ANCA ELISAs, or in at least four of the six MPO-ANCA ELISAs. Of the 26 sera that demonstrated cytoplasmic fluorescence (C-ANCA), 23 (89%) contained PR3-ANCA and three (11%) had MPO-ANCA. Two sera that were negative by indirect immunofluorescence (IIF) contained PR3-ANCA. Of the 26 sera with perinuclear fluorescence (P-ANCA), 19 (73%) contained MPO-ANCA, and one (4%) had PR3-ANCA. Six sera with P-ANCA did not have PR3-or MPO-ANCA. No serum that was negative by IIF contained MPO-ANCA. For the different PR3-ANCA ELISAs, sensitivities ranged from 88 to 100%, and specificities from 91 to 100%. For the MPO-ANCA ELISAs, sensitivities varied from 59 to 100% and specificities from 83 to 100%. The highest sensitivity and specificity for both the PR3-and MPO-ANCA ELISAs were obtained with the IBL and Eurodiagnostica assays. The in-house PR3-ANCA ELISA performed slightly less well than the commercial assays, but the performance of the in-house MPO-ANCA assay was comparable or better.
Anti-Neu5Gc and anti-non-Neu5Gc antibodies in healthy humans
PloS one, 2017
Our group previously investigated the levels of anti-Gal and anti-nonGal IgM and IgG in a cohort of 75 healthy humans of various backgrounds, and found some significant differences related to factors such as age, gender, ABO blood group, diet, vaccination history, and geographic location during childhood. We have now expanded our cohort (n = 84) to investigate the levels of anti-Neu5Gc and anti-nonGal/nonNeu5Gc antibodies in healthy humans. Anti-nonGal and anti-nonGal/nonNeu5Gc human IgM and IgG binding to pRBCs and pAECs from GTKO/CD46 and GTKO/CD46/Neu5GcKO pigs were measured by flow cytometry. Anti-Gal and anti-Neu5Gc IgM and IgG levels were measured by ELISA. In summary, (i) the great majority (almost 100%) of humans had anti-Neu5Gc IgM and IgG antibodies that bound to pAECs and approximately 50% had anti-Neu5Gc antibodies that bound to pRBCs, (ii) there was significantly less human antibody binding to pig cells that did not express either Gal or Neu5Gc compared with those that ...
A Rapid ELISA Test for Detection of Human Paraproteins
Clinical Chemistry and Laboratory Medicine, 1996
A rapid ELISA test for detection, characterization and quantification of human paraproteins was developed. The proposed method is a sandwich ELISA, where the capture antibody is specific for a given heavy chain (γ, α or μ) and the labelled antibody is specific either for κ or for λ light chain. Both standard and patient sera are tested with all six possible antibody combinations. Each paraprotein produces a significant increase in titre (as compared with standard) only when tested with the relevant pair of antibodies. This enables the determination of the isotype and light chain type of the paraprotein and the evaluation of its relative quantity in patient serum. The accuracy of the assay (relative deviation) varies from 0.04 for γλ to 0.19 for ακ. The cutoff values for each type of polyclonal immunoglobulin were determined with 200 healthy donor sera. 103 patient sera were analysed. ELISA data are in good agreement with M-component and other clinical data.
Clinical Chemistry and Laboratory Medicine, 2008
Background: The single nucleotide Marburg I (MRI) polymorphism of the factor VII-activating protease (FSAP) gene, the prourokinase-activating activity of FSAP, and antigen levels of FSAP in plasma have been associated with incidence and progression of carotid stenosis and venous thromboembolism. However, more information on the extent of these associations, potential further ones, and respective clinical utilities remain to be determined. At present, testing is performed mainly by PCR assays based on probes or SYBR Green I. Some studies include testing for antigen levels of total FSAP and its ability to activate prourokinase. To test large cohorts, it is beneficial to rely on assays that are cost-effective, reliable, easy to use, rapid to perform, and that may eventually be automated. In addition, it appears advantageous to use functional tests or tests that determine antigen levels as they may relate more closely to the phenotype than the genotype does. Methods: Tests for the measurements of antigen levels of FSAP and its prourokinase-activating activity were improved and performance characteristics assessed. To determine the FSAP genotypes, an amplification created restriction site (ACRS) PCR test was developed. Results: Key performance characteristics of the FSAP activity and antigen tests were as follows: measuring range: 350-1400 mPEU/mL and 1.8-120 ng/mL, total a We are deeply saddened by the death of our very much liked and highly esteemed colleague and friend Stefan Teigelkamp. Stefan died much too early at the age of 46 on June 5th, 2008. During his tenure with Siemens Healthcare Diagnostics, Stefan made numerous valuable contributions in the area of product development and improvement. He will be missed for his energetic leadership in research and development. We will always keep him in very good memory.