Design, Construction, and Two Applications for an Automated Flow—Cell Polarization Fluorometer with Digital Read Out: Enzyme-Inhibitor (Antitrypsin) Assay and Antigen—Antibody (Insulin—Insulin Antiserum) Assay

Abstract We have constructed and automated a flow cell polarization fluorometer and demonstrated two specific clinical applications of fluorescence polarization assay, i.e., the enzyme-inhibitor assay of antitrypsin in serum and the antigen-antibody assay of insulin and its antibody. A signal is displayed directly and is immediately available for chart recording and (or) digital data processing. Since fluorescence polarization offers a number of choices in assay parameters and the use of different fluorescent probes, its development for rapid simultaneous measurements of multiple components in body fluids should be possible.

Download Full-text

Fluorescence polarization assay improves the rapid detection of human brucellosis in China

Infectious Diseases of Poverty ◽

10.1186/s40249-021-00834-3 ◽

2021 ◽

Vol 10 (1) ◽

Author(s):

Shuai-Bing Dong ◽

Di Xiao ◽

Jing-Yao Liu ◽

Hui-Mei Bi ◽

Zun-Rong Zheng ◽

...

Keyword(s):

Fluorescence Polarization ◽

Clinical Symptoms ◽

Confirmatory Test ◽

Human Brucellosis ◽

Coincidence Rate ◽

Epidemiological Risk ◽

Kappa Value ◽

Fluorescence Polarization Assay ◽

Sensitivity Specificity ◽

Antigen Antibody

Abstract Background Brucellosis is an infectious-allergic zoonotic disease caused by bacteria of the genus Brucella. Early diagnosis is the key to preventing, treating, and controlling brucellosis. Fluorescence polarization immunoassay (FPA) is a new immunoassay for relatively rapid and accurate detection of antibodies or antigens based on antigen–antibody interaction. However, there is no report on FPA-based detection of human brucellosis in China. Therefore, this study is to evaluate the value of FPA for the diagnosis of human brucellosis in China. Methods We recruited 320 suspected brucellosis cases who had the clinical symptoms and epidemiological risk factors between January and December, 2019. According to China Guideline for Human Brucellosis Diagnosis, the Rose Bengal test (RBT) was used for the screening test, and the serum agglutination test (SAT) was used as the confirmatory test. Brucellosis was confirmed only if the results of both tests were positive. Additionally, FPA and enzyme linked immune sorbent assay (ELISA) were compared with SAT, and their sensitivity, specificity, coincidence rate and consistency coefficient (Kappa value) as diagnostic tests were analyzed individually and in combination. The optimal cut-off value of FPA was also determined using the receiver operator characteristic (ROC) curve. Results The optimum cut-off value of FPA was determined to be 88.5 millipolarization (mP) units, with a sensitivity of 94.5% and specificity of 100.0%. Additionally, the coincidence rate with the SAT test was 96.6%, and the Kappa value (0.9) showed excellent consistency. The sensitivity and specificity of FPA and ELISA combined were higher at 98.0% and 100.0% respectively. Conclusions When the cut-off value of FPA test is set at 88.5 mP, it has high value for the diagnosis of brucellosis. Additionally, when FPA and ELISA are combined, the sensitivity of diagnosis is significantly improved. Thus, FPA may have potential in the future as a diagnostic method for human brucellosis in China. Graphic abstract

Download Full-text

Development of a fluorescence polarization assay to screen for inhibitors of the FtsZ/ZipA interaction

Analytical Biochemistry ◽

10.1016/j.ab.2003.08.033 ◽

2003 ◽

Vol 323 (2) ◽

pp. 224-233 ◽

Cited By ~ 49

Author(s):

Cynthia Hess Kenny ◽

Weidong Ding ◽

Kerry Kelleher ◽

Susan Benard ◽

Elizabeth Glasfeld Dushin ◽

...

Keyword(s):

Fluorescence Polarization ◽

Fluorescence Polarization Assay

Download Full-text

Efficient and Expedient Two-Step Pyranose-Retaining Fluorescein Conjugation of Complex Reducing Oligosaccharides: Galectin Oligosaccharide Specificity Studies in a Fluorescence Polarization Assay

Bioconjugate Chemistry ◽

10.1021/bc034130j ◽

2003 ◽

Vol 14 (6) ◽

pp. 1289-1297 ◽

Cited By ~ 18

Author(s):

Christopher T. Öberg ◽

Susanne Carlsson ◽

Eric Fillion ◽

Hakon Leffler ◽

Ulf J. Nilsson

Keyword(s):

Fluorescence Polarization ◽

Fluorescence Polarization Assay

Download Full-text

AUTOMATIC EEG TAPE-COMPUTER SYSTEM FOR ANALOG-DIGITAL DATA PROCESSING OF FREQUENCY-SPECTRAL INFORMATION

Acta Neurologica Scandinavica ◽

10.1111/j.1600-0404.1966.tb04766.x ◽

1966 ◽

Vol 42 (S22) ◽

pp. 5-19 ◽

Cited By ~ 1

Keyword(s):

Data Processing ◽

Computer System ◽

Digital Data ◽

Spectral Information ◽

Digital Data Processing

Download Full-text

A Fluorescence Polarization Assay for Cyclic Nucleotide Phosphodiesterases

CrossRef Listing of Deleted DOIs ◽

10.1177/108705710200700305 ◽

2002 ◽

Vol 7 (3) ◽

pp. 215-222 ◽

Cited By ~ 38

Author(s):

Wei Huang ◽

Yan Zhang ◽

J. Richard Sportsman

Keyword(s):

Cyclic Nucleotide ◽

Fluorescence Polarization ◽

Drug Targets ◽

Second Messengers ◽

Reaction Products ◽

Cyclic Nucleotide Phosphodiesterases ◽

Extracellular Signals ◽

Fluorescence Polarization Assay ◽

Hydrolysis Of ◽

Nucleotide Phosphodiesterases

Cyclic nucleotide phosphodiesterases (PDEs) catalyze the hydrolysis of the 3′-ester bond of cyclic AMP (cAMP) and cyclic GMP (cGMP), important second messengers in the transduction of a variety of extracellular signals. There is growing interest in the study of PDEs as drug targets for novel therapeutics. We describe the development of a homogeneous fluorescence polarization assay for PDEs based on the strong binding of PDE reaction products (i.e., AMP or GMP) onto modified nanoparticles through interactions with immobilized trivalent metal cations. This assay technology (IMAP) is applicable to both cAMP- and cGMP-specific PDEs. Results of the assay in 384- and 1536-well microplates are presented.

Download Full-text