scholarly journals Development of a highly sensitive chemiluminescent enzyme immunoassay for fragmented cytokeratin 18 using new antibodies

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Minori Yamada ◽  
Akiko Eguchi ◽  
Koji Okuno ◽  
Koji Sakaguchi ◽  
Tetsuji Yamaguchi
Keyword(s):  
Enzyme Immunoassay ◽  
Detection Sensitivity ◽  
Amino Acid Residues ◽  
Cytokeratin 18 ◽  
E Coli ◽  
Coefficients Of Variation ◽  
Highly Sensitive ◽  
Clinical Diagnoses ◽  
Analysis Of Performance ◽  
Chemiluminescent Enzyme Immunoassay

AbstractFragmented cytokeratin 18 (fCK18) released from epithelial cells undergoing apoptosis is widely studied in various diseases. However, fCK18 measurement is not utilized in clinical practice due to imprecise disease-state cutoff values. Therefore, we set out to generate new monoclonal antibodies (mAbs) and a recombinant fCK18 (rfCK18) calibrator in an effort to develop a highly sensitive chemiluminescent enzyme immunoassay (CLEIA). New capture mAb (K18-624) had a high binding ability compared to the current commercial antibody. New detection mAb (K18-328) recognized 323S-340G of CK18. A rfCK18 was expressed in the soluble fraction of E. coli when the N-terminal region (260 amino acid residues) of CK18 was truncated. Analysis of performance and measurement of human fCK18 were evaluated using K18-624 and K18-328 in a highly sensitive CLEIA. The coefficients of variation (CV) for within-run and between-day repeatability were below 10% and the recoveries were in the range of 15%. The detection sensitivity was 0.056 ng/mL. Serum fCK18 levels were significantly increased in non-alcoholic steatohepatitis (NASH) patients when compared to healthy individuals. Our new fCK18 mAbs showed high affinity and sensitivity. CLEIA using our new antibodies will be useful in measuring fCK18 in human blood thereby generating accurate clinical diagnoses of human liver diseases.

Specific enzyme immunoassay for human chorionic gonadotrophin

1981 ◽  
Vol 97 (4) ◽  
pp. 562-568 ◽  
Author(s):  
Tatsuhiro Sekiya ◽  
Yoshihito Furuhashi ◽  
Setsuko Goto ◽  
Shigeaki Kaseki ◽  
Yutaka Tomoda ◽  
...  
Keyword(s):  
Enzyme Immunoassay ◽  
Solid Phase ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Serum Samples ◽  
Coefficients Of Variation ◽  
Highly Sensitive ◽  
Assay Tube ◽  
Sandwich Type ◽  
Serial Samples

Abstract. A sandwich-type enzyme immunoassay system specific for human chorionic gonadotrophin (hCG) was prepared with the antibody Fab'-β-D-galactosidase complex and the antibody F(ab')2-immobilized silicone rubber solid phase by using a purified antibody to β subunit of hCG (hCGβ). The assay system cross-reacted less than 4% with human luteinizing hormone (hLH) and human follicular stimulating hormone (hFSH), and proved to be highly sensitive with hCG measurable at levels as low as 0.3 mIU per assay tube. Using 50 μl of serum sample, 6–600 mIU/ml of hCG in serum could be determined specifically with the same degree of precision as in radioimmunoassay but without sample interference with the assay. The coefficients of variation within-run and between-run were 8.6–8.9%, and 4.9– 10.7%, respectively. Values obtained with the enzyme immunoassay correlated well with those of radioimmunoassay ([unk] = 0.98, slope = 0.94, y-intercept = 10.2 mIU/ml for 75 serum samples). Results of the immunoassay of hCG levels in serial samples of serum from healthy women and patients with choriocarcinoma show that this method is useful in the clinical diagnosis of trophoblastic disease.

Highly Sensitive Sandwich Enzyme Immunoassay for Alpha-Fetoprotein in Human Saliva

1992 ◽  
Vol 29 (5) ◽  
pp. 519-522 ◽  
Author(s):  
Xian Yang Yio ◽  
Jing Jiang ◽  
Fen Zhi Yin ◽  
Ke-He Ruan
Keyword(s):  
Enzyme Immunoassay ◽  
Human Saliva ◽  
Minimum Detectable Concentration ◽  
Igg Antibody ◽  
Coefficients Of Variation ◽  
Highly Sensitive ◽  
Detectable Concentration ◽  
Polystyrene Plate ◽  
Normal Human ◽  
Sandwich Enzyme Immunoassay

To determine α-fetoprotein (AFP) in human saliva, a highly sensitive sandwich enzyme immunoassay for saliva AFP was developed. AFP standards and saliva samples were added into the wells of a polystyrene plate coated with goat IgG antibody against human AFP. After incubation, the wells were washed and horseradish peroxidase-labelled antibody was added. The enzyme activity specifically bound to the well was assayed using 3,3′,5,5′-tetramethylbenzidine and hydrogen peroxide as substrate. The reaction was stopped by addition of 2 M sulphuric acid and the AFP concentration was determined from the absorbance at 450 nm. The minimum detectable concentration was 8 ng/L. The recovery of AFP mixed with human saliva was 91·1–102·4%. The within-assay and between-assay coefficients of variation were 6·5–8·9% and 7·6–10·8%, respectively. The assay correlated well with a radioimmunoassay for human AFP ( r = 0·985, n = 13, P < 0·001). The mean concentration of AFP in normal human saliva was 14·3 ng/L (SEM = 4·9 ng/L, n = 10) and significantly higher levels of saliva AFP were observed in hepatocellular carcinoma patients with positive serum AFP (mean 1367·8 ng/L, SEM 595·4 ng/L, n = 6; P < 0·001). Strong correlation was observed between saliva AFP and serum AFP ( r = 0·978, P < 0·01, n = 13).

Highly Sensitive Chemiluminescent Enzyme Immunoassay with Gelatin-Coated Ferrite Solid Phase

1994 ◽  
Vol 40 (9) ◽  
pp. 1830-1831 ◽  
Author(s):  
Mitsuo Isomura ◽  
Masayoshi Ueno ◽  
Kazuya Shimada ◽  
Hiroyuki Kogaki ◽  
Yoshihiro Ashihara
Keyword(s):  
Enzyme Immunoassay ◽  
Solid Phase ◽  
Highly Sensitive ◽  
Chemiluminescent Enzyme Immunoassay

Production of Antibodies and Development of Enzyme Immunoassay for Determination of Monensin in Biological Samples

1987 ◽  
Vol 70 (2) ◽  
pp. 201-205
Author(s):  
Michael E Mount ◽  
Douglas L Failla
Keyword(s):  
Enzyme Immunoassay ◽  
Immunosorbent Assay ◽  
Biological Samples ◽  
Enzyme Linked Immunosorbent Assay ◽  
Coefficients Of Variation ◽  
Highly Sensitive

Abstract Monensin is converted to monensin bromoacetate, which provides successful coupling to protein and allows production of monensinspeciflc rabbit antisera. A modified indirect enzyme-linked immunosorbent assay (ELISA) was developed, which is highly sensitive (2 ng/mL) to monensin determination. Monensin recovery was 53- 81% at 10 ppb in sera or urine, and 81-130% at 100 ppb in dichloromethane- extracted feces or water. Overall recovery was 98.8% with coefficients of variation from 6 to 52% over the range of monensin concentrations studied. This is the first immunoassay reported for the carboxylic ionophore antibiotics.

scholarly journals Evaluation of the highly sensitive chemiluminescent enzyme immunoassay “Lumipulse HBsAg-HQ” for hepatitis B virus screening

2017 ◽  
Vol 32 (4) ◽  
pp. e22334 ◽  
Author(s):  
Matsuo Deguchi ◽  
Masanori Kagita ◽  
Nori Yoshioka ◽  
Hiroko Tsukamoto ◽  
Miyuki Takao ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Enzyme Immunoassay ◽  
Highly Sensitive ◽  
B Virus ◽  
Chemiluminescent Enzyme Immunoassay ◽  
Virus Screening

scholarly journals Comparison of a Miniaturized Cassette PCR System with a Commercially Available Platform for Detecting Escherichia coli in Beef Carcass Swabs

Micromachines ◽  
2021 ◽  
Vol 12 (8) ◽  
pp. 959
Author(s):  
Dammika P. Manage ◽  
Jana Lauzon ◽  
Linda M. Pilarski ◽  
Patrick M. Pilarski ◽  
Lynn M. McMullen
Keyword(s):  
Escherichia Coli ◽  
Curve Analysis ◽  
Detection Sensitivity ◽  
Conventional Pcr ◽  
False Negatives ◽  
Melt Curve Analysis ◽  
E Coli ◽  
Highly Sensitive ◽  
Multiple Samples ◽  
Beef Carcass

Detection sensitivity of cassette PCR was compared with a commercial BAX® PCR system for detection of eae and stx genes in Escherichia coli from 806 beef carcass swabs. Cassette PCR detects multiple genetic markers on multiple samples using PCR and melt curve analysis. Conventional PCR served as a gold standard. Overall, for positive and negative concordance, cassette PCR was 98.6% concordant with conventional PCR, and BAX PCR was 65.4% concordant. Of 806 beef carcass swabs, 339 by cassette PCR and 84 by BAX PCR harbored eae + stx+E. coli. For BAX PCR reactions, 84% of eae+ swabs, 79% of stx+ swabs, and 86% of eae + stx+ swabs were also detected by cassette PCR. For cassette PCR reactions, 457 swabs were eae+ with only 117 scored as eae+ using BAX PCR for 26% positive concordance. For stx primers, cassette PCR scored 480 samples as stx+ but only 215 samples were stx+ by BAX PCR, giving 45% positive concordance. Importantly, cassette PCR scored 339 swabs as harboring eae + stx+ E. coli, but BAX PCR detected only 71 positives giving only 21% positive concordance, with many false negatives. Cassette PCR is a highly sensitive method for detection of STEC genes in E. coli found in carcass swabs.

Highly sensitive phage-magnetic-chemiluminescent enzyme immunoassay for determination of zearalenone

2020 ◽  
Vol 325 ◽  
pp. 126905 ◽  
Author(s):  
Ruxia Liu ◽  
Ruirui Shi ◽  
Wenting Zou ◽  
Wenhua Chen ◽  
Xianchao Yin ◽  
...  
Keyword(s):  
Enzyme Immunoassay ◽  
Highly Sensitive ◽  
Chemiluminescent Enzyme Immunoassay
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