Particle concentration fluorescence immunoassay: a new immunoassay technique for quantification of human immunoglobulins in serum.

1985 ◽  
Vol 31 (9) ◽  
pp. 1487-1490 ◽  
Author(s):  
C MacCrindle ◽  
K Schwenzer ◽  
M E Jolley
Keyword(s):  
Particle Concentration ◽  
Solid Phase ◽  
Front Surface ◽  
Human Antibody ◽  
Fluorescence Immunoassay ◽  
Polystyrene Spheres ◽  
Human Immunoglobulins ◽  
Immunoassay Technique ◽  
Total Particle ◽  
Sandwich Assays

Abstract A new fluorescence immunoassay technique, particle concentration fluorescence immunoassay (PCFIA), has been developed for quantifying the human immunoglobulins (IgA, IgM, and IgG). In these "two-site sandwich assays," the capture antibody is immobilized on small polystyrene spheres and the tracer is fluorescein-labeled antibody. Polystyrene particles less than 1 microm in diameter make up the solid phase, to which goat anti-human antibody for each respective assay is attached. Serum specimens are diluted (5000-fold for IgA or IgM, 20 000-fold for IgG) placed on the 96-well Pandex assay plate; and mixed with the solid phase and tracer (fluorescein-labeled goat anti-human IgA, IgM, or IgG), which are added automatically by the Pandex Screen Machine. This instrument incubates the reaction mixture for 17 min at ambient temperature, separates the bound and free label by filtration, washes the solid phase, and determines the total particle-bound fluorescence by front-surface fluorimetry or epifluorescence, calculates results, and generates detailed reports. Ninety-six specimens may be analyzed in 29 min or 960 specimens in 136 min. Results by PCFIA for IgA, IgM, and IgG in serum correlated well with those by rate nephelometry.

Particle concentration fluorescence immunoassay (PCFIA): a new, rapid immunoassay technique with high sensitivity

1984 ◽  
Vol 67 (1) ◽  
pp. 21-35 ◽  
Author(s):  
Michael E. Jolley ◽  
Chao-Huei J. Wang ◽  
Steven J. Ekenberg ◽  
Mark S. Zuelke ◽  
David M. Kelso
Keyword(s):  
Particle Concentration ◽  
High Sensitivity ◽  
Fluorescence Immunoassay ◽  
Immunoassay Technique

scholarly journals Simultaneous Determination of Different Antibiotic Residues in Bovine and in Porcine Kidneys by Solid-Phase Fluorescence Immunoassay

2003 ◽  
Vol 86 (2) ◽  
pp. 236-240 ◽  
Author(s):  
Lieve Okerman ◽  
Katia De Wasch ◽  
Jan Van Hoof ◽  
Walter Smedts
Keyword(s):  
Solid Phase ◽  
Kidney Tissue ◽  
Test System ◽  
Penicillin G ◽  
Fluorescence Immunoassay ◽  
Beta Lactam ◽  
Residue Detection ◽  
Beta Lactam Antibiotics ◽  
Minimal Sample

Abstract Parallux®, a solid-phase fluorescence immunoassay (SPFIA) developed for antibiotic residue detection in milk, was used for analysis of bovine and porcine kidney tissue. Four tetracyclines, 2 broad-spectrum cephalosporins, 3 beta-lactam antibiotics, and cephapirin were detected in one run after minimal sample preparation. This commercially available test system is designed as cartridges, each with a combination of 1–4 tests. One cartridge can be used to detect 4 analytes in the same sample, or 1 or 2 analytes in different samples. The cartridge with the combination tetracyclines–ceftiofur–penicillin–cephapirin was selected because tetracyclines, beta-lactam antibiotics as well as cephalosporins, are registered for oral or parenteral use in bovines and pigs in Europe. The test is qualitative and is recommended only for screening. Tetracycline, oxytetracycline, chlortetracycline, and doxycycline were easily detected at 300 ppb with the tetracyclines channel; ceftiofur at 1000 ppb and cefquinome at 200 ppb with the ceftiofur channel; penicillin G, ampicillin, and amoxicillin at 50 ppb with the penicillin channel; and cephapirin at 100 ppb with the cephapirin channel. These levels are equal to or lower than the corresponding maximal residue limits in kidney tissue. Cephalexin was not detected. The SPFIA test can be used as an alternative to classical inhibition tests and for post-screening inhibitor- positive kidneys, because it detects 3 specific groups of antibiotics, which enables selection of specific confirmatory methods for identification and quantification.

Application of a solid-phase fluorescence immunoassay to determine amoxicillin residues in fish tissue

2010 ◽  
Vol 58 (1) ◽  
pp. 83-89 ◽  
Author(s):  
Eun-Kee Park ◽  
Won Jung ◽  
Hu-Jang Lee
Keyword(s):  
Solid Phase ◽  
Paralichthys Olivaceus ◽  
Fish Tissue ◽  
Sea Bream ◽  
Sebastes Schlegeli ◽  
Withdrawal Period ◽  
Fluorescence Immunoassay ◽  
Red Sea Bream ◽  
Pagrus Major ◽  
Farmed Fish

The present study demonstrates an application of Parallux™ (a solid-phase fluorescence immunoassay) for amoxicillin analysis in fish tissue. Amoxicillin at the recommended therapeutic dose (400 mg/kg body weight) was orally administered to three groups of 25 olive flounder (Paralichthys olivaceus), 25 rockfish (Sebastes schlegeli) and 25 red sea bream (Pagrus major) for 7 consecutive days. Amoxicillin was detected in the muscle of fish treated by the 3rd day of the withdrawal period. The recovery rates of all spiked muscle samples were > 86% of the spiked values. The present study showed that solid-phase fluorescence immunoassay can be easily adopted in predicting amoxicillin residues in the muscle tissue of farmed fish.

Bacteria as solid phase in a concentration fluorescence immunoassay analysis of antibodies to surface antigens

1990 ◽  
Vol 126 (2) ◽  
pp. 247-252 ◽  
Author(s):  
William R. Schwan ◽  
Carl Waltenbaugh ◽  
James L. Duncan
Keyword(s):  
Solid Phase ◽  
Surface Antigens ◽  
Fluorescence Immunoassay ◽  
As Solid Phase

The specificity of a solid phase radioimmunoassay for human immunoglobulins

1975 ◽  
Vol 7 (2-3) ◽  
pp. 179-185 ◽  
Author(s):  
R.P. Eady ◽  
J.C. Chapple ◽  
D.W. Hough ◽  
G.T. Stevenson
Keyword(s):  
Solid Phase ◽  
Human Immunoglobulins ◽  
Solid Phase Radioimmunoassay

Development and application of monoclonal antibodies to human cardiac myoglobin in a rapid fluorescence immunoassay

1991 ◽  
Vol 37 (8) ◽  
pp. 1356-1364 ◽  
Author(s):  
D P Silva ◽  
Y Landt ◽  
S E Porter ◽  
J H Ladenson
Keyword(s):  
Monoclonal Antibodies ◽  
Particle Concentration ◽  
High Dose ◽  
Fluorescence Immunoassay ◽  
Hook Effect ◽  
One Step ◽  
Analytical Importance ◽  
Rapid Kinetics ◽  
Kinetics Of Reaction ◽  
Kinetics Of

Abstract Myoglobin (Mb) is considered a useful marker for early detection of myocardial infarction and for monitoring cardiac reperfusion after thrombolytic therapy. We developed eight monoclonal antibodies to human cardiac Mb, characterized their epitopic reactivity, and determined which combinations of the antibodies are useful in two-site immunoassays. We configured two of the monoclonal antibodies in a one-step, two-site particle concentration fluorescence immunoassay (PCFIA) for measurement of Mb. The PCFIA has rapid kinetics of reaction, being complete in 15 min, and has a linear analytical range of 20-675 micrograms/L for human Mb. Although the PCFIA has a high-dose "hook" effect, this is of no analytical importance at concentrations of Mb less than or equal to 148,000 micrograms/L. The assay is not subject to interference from icterus (bilirubin less than or equal to 360 mg/L), has no cross-reaction with hemoglobin (less than or equal to 42 g/L), and may be performed with either plasma or serum in approximately 1 h. The intra- and interassay imprecisions (CV) of the method are less than 10% for concentrations of Mb within the normal range and less than 4% at higher concentrations. A comparison of the PCFIA with a commercial radioimmunoassay showed that results of the two assays correlate well (PCFIA = 0.88 x RIA + 18, r = 0.990, n = 171).

Ultrasensitive Thyrotropin Immunoassay Based on Enzymatically Amplified Time-Resolved Fluorescence with a Terbium Chelate

1992 ◽  
Vol 38 (4) ◽  
pp. 545-548 ◽  
Author(s):  
A Papanastasiou-Diamandi ◽  
T K Christopoulos ◽  
E P Diamandis
Keyword(s):  
Alkaline Phosphatase ◽  
Detection Limit ◽  
Solid Phase ◽  
Thyroid Stimulating Hormone ◽  
Fluorescence Immunoassay ◽  
Time Resolved Fluorescence ◽  
Time Resolved ◽  
Highly Sensitive ◽  
Assay Time ◽  
Highly Sensitive Detection

Abstract We describe an ultrasensitive, enzymatically amplified time-resolved fluorescence immunoassay of thyrotropin (thyroid-stimulating hormone) in serum with use of a terbium chelate as the detectable moiety. In this assay, thyrotropin is first simultaneously reacted with a solid-phase (microtiter well) monoclonal antibody and a soluble biotinylated monoclonal detection antibody. After washing, a streptavidin-alkaline phosphatase conjugate is added, followed by another washing. Alkaline phosphatase acts on the substrate 5-fluorosalicyl phosphate (FSAP) to produce 5-fluorosalicylic acid (FSA). FSA, but not FSAP, can then form with Tb3+ and EDTA a highly fluorescent ternary complex of long fluorescence lifetime. This complex is quantified with time-resolved fluorometry. The thyrotropin assay is highly sensitive (detection limit approximately 0.003 milli-int. unit/L when a total assay time of 85 min is used), precise, and accurate. The thyrotropin assay can also be completed in less than 30 min (detection limit 0.013 milli-int. unit/L), thus making this procedure a candidate technology for high-throughput automated analyzers.

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