An enzyme immunoassay for galactosyltransferase isoenzyme II, and its clinical application to cancer diagnosis

1990 ◽  
Vol 36 (4) ◽  
pp. 598-601 ◽  
Author(s):  
M Uemura ◽  
M Yamasaki ◽  
S Yoshida ◽  
T Uezima ◽  
T Sakaguchi ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Esophageal Cancer ◽  
Enzyme Immunoassay ◽  
Cancer Diagnosis ◽  
Blood Type ◽  
Benign Diseases ◽  
Standard Curve ◽  
Color Development ◽  
Sandwich Type ◽  
Normal Controls

Abstract In this "sandwich"-type enzyme immunoassay of galactosyltransferase isoenzyme II (GT-II), we used the monoclonal antibody MAb 3872 previously established and characterized to be specific for GT-II. Plastic beads coated with MAb 3872 were incubated with serum and buffer, washed, then incubated with MAb 3872 conjugated with horseradish peroxidase and again washed. Peroxidase activity remaining on the beads was measured by color development with o-phenylenediamine. The assay standard curve was linear from 0 to 150 kU/L. Inter- and intra-assay CVs were less than 9% and less than 7%, respectively. Mean GT-II values for 370 normal controls were 6.8 (SD 3.4) kU/L with no significant sex-, smoking-, or blood-type-related differences. We also assayed serum from patients with 11 different cancers or with matched benign diseases. The diagnostic sensitivity of GT-II for liver and esophageal cancer was 0.91 and 0.72, respectively, apparently higher than for carcinoembryonic antigen (CEA) or alpha-fetoprotein. GT-II and CEA concentrations in sera were not correlated. Evidently this assay may be useful as a cancer diagnostic test to supplement existing serological methods.

Sandwich enzyme immunoassay of osteocalcin in serum with use of an antibody against human osteocalcin

1993 ◽  
Vol 39 (6) ◽  
pp. 942-947 ◽  
Author(s):  
D A Monaghan ◽  
M J Power ◽  
P F Fottrell
Keyword(s):  
Monoclonal Antibody ◽  
Enzyme Immunoassay ◽  
Solid Phase ◽  
Sandwich Elisa ◽  
Normal Subjects ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Standard Curve ◽  
Microtiter Plates ◽  
Human Osteocalcin

Abstract We have developed and thoroughly validated a solid-phase sandwich enzyme-linked immunosorbent assay (ELISA) on microtiter plates for osteocalcin in human serum with use of an antibody raised against human osteocalcin. We used a monoclonal antibody against bovine osteocalcin as the capture antibody; the second antibody was a polyclonal antibody against human osteocalcin. The amount of bound second antibody was determined with use of swine anti-rabbit antibody labeled with horseradish peroxidase. We demonstrated independence of volume and determined the recovery of added standard and within- and between-assay precision. The minimal detection limit for osteocalcin was between 1.0 and 1.5 micrograms/L and the midpoint of the standard curve ranged from 14 to 17 micrograms/L. The intraassay CV was < or = 8% in the range 2.7-52 micrograms/L; the interassay CV was usually < or = 15% in the same range. Analytical recovery of human osteocalcin standard added to serum samples was consistently > 90%. Values for osteocalcin measured in serum from 44 normal subjects were similar to those obtained with a competitive enzyme immunoassay (EIA) that used a monoclonal antibody against bovine osteocalcin. There was a good correlation between the two assays [r2 = 0.877, slope and intercept (+/- SE) = 0.88(+/- 0.051) and 0.316(+/- 0.523), respectively]. The range and mean (+/- SD) for the sandwich ELISA and the competitive EIA were 1.7-18.1 micrograms/L [8.7(+/- 4.4) micrograms/L] and 1.9-22.8 micrograms/L [9.1(+/- 4.4) micrograms/L], respectively.

scholarly journals Potential miRNA biomarkers for the diagnosis and prognosis of esophageal cancer detected by a novel absolute quantitative RT-qPCR method

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Zhiyuan Lin ◽  
Yongquan Chen ◽  
Yanling Lin ◽  
Huayue Lin ◽  
Hongwei Li ◽  
...  
Keyword(s):  
Esophageal Cancer ◽  
High Sensitivity ◽  
Benign Diseases ◽  
Diagnosis And Prognosis ◽  
Qpcr Method ◽  
Sensitivity Specificity ◽  
Combined Detection ◽  
Normal Controls ◽  
Potential Biomarkers ◽  
High Application

AbstractmiRNAs are expected to become potential biomarkers in the diagnosis and prognosis of Esophageal cancer (EC). Through a series of screening, miR-34a-5p, miR-148a-3p and miR-181a-5p were selected as EC-associated miRNAs. Based on AllGlo probe, a novel absolute quantitative RT-qPCR method with high sensitivity, specificity and accuracy was established for detecting miRNAs. Then the clinical significance of these 3 miRNAs was explored with 213 patients (166 cases with EC and 47 cases with benign diseases) and 170 normal controls. Compared with normal controls, the level of miR-34a-5p increased while miR-148a-3p and miR-181a-5p decreased in EC and benign patients (P < 0.001), and the level of miR-181a-5p in early EC patients was significantly lower (P < 0.001). According to logistic regression analysis, combined detection of miR-34a-5p, miR-148a-3p and Cyfra21-1 provided the highest diagnosis efficiency of 85.07% with sensitivity and specificity reaching 85.45% and 84.71%. Compared with preoperative samples, the level of miR-34a-5p decreased while miR-148a-3p and miR-181a-5p increased in postoperative samples (P < 0.001). Collectively, this first developed, novel absolute quantitative RT-qPCR method exhibits high application value in detecting miRNAs, miR-34a-5p, miR-148a-3p and miR-181a-5p may serve as potential biomarkers in the diagnosis and prognosis of EC, and miR-181a-5p probably could serve as a new biomarker for early EC.

A Monoclonal Antibody-Based Enzyme Immunoassay for Fibrin Degradation Products in Plasma

1988 ◽  
Vol 59 (02) ◽  
pp. 310-315 ◽  
Author(s):  
P W Koppert ◽  
E Hoegee-de Nobel ◽  
W Nieuwenhuizen
Keyword(s):  
Enzyme Immunoassay ◽  
Degradation Products ◽  
Blood Clot ◽  
Tissue Type ◽  
Fibrin Degradation Products ◽  
Type Plasminogen Activator ◽  
Strong Positive Reaction ◽  
Sandwich Type ◽  
Capture Antibody

SummaryWe have developed a sandwich-type enzyme immunoassay (EIA) for the quantitation of fibrin degradation products (FbDP) in plasma with a time-to-result of only 45 minutes.* The assay is based on the combination of the specificities of two monoclonal antibodies (FDP-14 and DD-13), developed in our institute. FDP-14, the capture antibody, binds both fibrinogen degradation products (FbgDP) and FbDP, but does not react with the parent fibrin(ogen) molecules. It has its epitope in the E-domain of the fibrinogen molecule on the Bβ-chain between amino acids 54-118. Antibody DD-13 was raised using D-dimer as antigen and is used as a tagging antibody, conjugated with horse-radish peroxidase. A strong positive reaction is obtained with a whole blood clot lysate (lysis induced by tissue-type plasminogen activator) which is used as a standard. The EIA does virtually not detect FbgDP i. e. purified fragments X, Y, or FbgDP generated in vitro in plasma by streptokinase treatment. This indicates that the assay is specific for fibrin degradation products.We have successfully applied this assay to the plasma of patients with a variety of diseased states. In combination with the assay previously developed by us for FbgDP and for the total amount of FbgDP + FbDP (TDP) in plasma, we are now able to study the composition of TDP in patients plasma in terms of FbgDP and FbDP.

Development of a Monoclonal Antibody-Based Enzyme Immunoassay for the Pyrethroid Insecticide Deltamethrin

2010 ◽  
Vol 58 (14) ◽  
pp. 8189-8195 ◽  
Author(s):  
Ye Kong ◽  
Qi Zhang ◽  
Wen Zhang ◽  
Shirley J. Gee ◽  
Peiwu Li
Keyword(s):  
Monoclonal Antibody ◽  
Enzyme Immunoassay ◽  
Pyrethroid Insecticide

scholarly journals Bound/free separation methods in steroid enzyme immunoassay with monoclonal antibody.

1988 ◽  
Vol 36 (9) ◽  
pp. 3525-3531 ◽  
Author(s):  
HIROSHI HOSODA ◽  
REIKO TSUKAMOTO ◽  
SAKIKO TAMURA ◽  
TOSHIO NAMBARA
Keyword(s):  
Monoclonal Antibody ◽  
Enzyme Immunoassay

Quantitative Assessment of the BCR-ABL Transcript Using the Cepheid Xpert BCR-ABL Monitor Assay

2007 ◽  
Vol 131 (6) ◽  
pp. 947-950
Author(s):  
Scott D. Dufresne ◽  
Dorothy R. Belloni ◽  
Norman B. Levy ◽  
Gregory J. Tsongalis
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcriptase ◽  
Coefficient Of Variation ◽  
Quantitative Assessment ◽  
Myelogenous Leukemia ◽  
Chain Reaction ◽  
Normal Peripheral Blood ◽  
Standard Curve ◽  
Polymerase Chain ◽  
Normal Controls

Abstract Context.—Chronic myelogenous leukemia (CML) and the assessment of the BCR-ABL transcript has become a new paradigm. Novel tyrosine kinase inhibitors as mainstream therapeutic options for the CML patient warrant routine quantification of the BCR-ABL transcript. The Xpert BCR-ABL Monitor assay is a nested reverse transcriptase polymerase chain reaction that greatly reduces technical time by using a single cartridge to isolate RNA and run a quantitative reverse transcriptase polymerase chain reaction. Objective.—To evaluate the Xpert BCR-ABL Monitor assay for quantitative assessment of the BCR-ABL transcript in CML patients. Design.—A standard curve of K-562 cells diluted in normal peripheral blood was used to test the sensitivity, linearity, and percent coefficient of variation of the assay. Specimen stability was tested by running standard curves immediately and after 24 hours or 96 hours of storage at 4°C. Specimens from normal controls, patients known to have CML, or patients suspected of having CML were also tested. Results.—The sensitivity of the assay was sufficient to detect 1 K-562 cell in 105 normal cells. The R2 of the standard curve was 0.98 and the percent coefficient of variation for each data point was 15% to 24%. Eleven of 14 patients with known CML on imatinib treatment tested positive for the BCR-ABL transcript, whereas 10 normal controls tested negative. Conclusions.—The Xpert BCR-ABL Monitor assay is a rapid, sensitive method for monitoring the presence of the BCR-ABL transcript in CML patients. The single-use cartridge minimizes hands-on technical time, minimizes the potential for contamination, and allows quantitative BCR-ABL testing to be performed in a random access fashion.

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