Direct solid-phase chemiluminescence immunoassay for salivary progesterone.

1986 ◽  
Vol 32 (5) ◽  
pp. 763-767 ◽  
Author(s):  
J De Boever ◽  
F Kohen ◽  
D Vandekerckhove
Keyword(s):  
Detection Limit ◽  
Luteal Phase ◽  
Solid Phase ◽  
Free Ligand ◽  
Follicular Phase ◽  
Chemiluminescence Immunoassay ◽  
Coefficients Of Variation ◽  
Analytical Recovery ◽  
Unstimulated Saliva ◽  
Direct Solid

Abstract A simple, direct chemiluminescence immunoassay for progesterone in mixed, unstimulated saliva is described. We use purified polyclonal anti-progesterone antibodies covalently coupled to polyacrylamide beads and progesterone-11 alpha-hemisuccinyl-aminobutylethyl isoluminol as the chemiluminescent ligand marker. Bound and free ligand are separated by simple centrifugation. The detection limit of the assay is 1.5 pg per tube (38 pmol/L). Intra- and interassay coefficients of variation for low, medium, and high progesterone concentrations are 7.9, 6.8, 8.8% and 9.3, 6.9, 8.5%, respectively. Analytical recovery of added progesterone is 99.6%. Mean +/- SD progesterone concentrations (pmol/L) in saliva are 178 +/- 46 in the follicular phase, 313 +/- 90 in the periovulatory phase, and 658 +/- 166 in the luteal phase of the menstrual cycle. Correlation between progesterone concentrations in serum (assayed by RIA after extraction) and in saliva is good (r = 0.88, p less than 0.001, n = 96). The assay is simple, fast (4 h, including 1.5 h of incubation time), and reliable.

Direct chemiluminescence immunoassay of estradiol in saliva

1990 ◽  
Vol 36 (12) ◽  
pp. 2036-2041 ◽  
Author(s):  
J De Boever ◽  
F Kohen ◽  
J Bouve ◽  
D Leyseele ◽  
D VandeKerckhove
Keyword(s):  
Detection Limit ◽  
Incubation Time ◽  
Solid Phase ◽  
Chemiluminescence Immunoassay ◽  
Microtiter Plates ◽  
Analytical Recovery ◽  
Plate Reader ◽  
Steroid Binding ◽  
Method R ◽  
Direct Solid

Abstract A sensitive and simple direct solid-phase chemiluminescence immunoassay is described for estradiol in saliva. In this assay, a second antibody is bound to the wells of microtiter plates. Either buffer with standards or saliva (100 microL) is incubated in these wells with monoclonal anti-estradiol antibody and with estradiol-isoluminol conjugate. Incubation time is 2 h. Chemiluminescence of the bound fraction is measured in a manually operated luminometer (Biocounter). The assay has a detection limit of 3.8 pmol/L; analytical recovery of added estradiol is 96.8% (SD 7.0%); within- and between-assay CVs range between 2.5% and 12.7%. Forty unknown saliva samples can be assayed and results calculated within 4.5 h. Results of a slightly modified procedure-with black microtiter plates and a prototype of an automated plate reader (Luminoskan)--compare well with those of the described method (r = 0.97). Because steroid-binding globulins have been found in saliva, the effect of displacing agents on the results of the direct chemiluminescence assay is described.

Evaluation of a direct solid-phase radioimmunoassay for progesterone, useful for monitoring luteal function.

1984 ◽  
Vol 30 (2) ◽  
pp. 284-286 ◽  
Author(s):  
N P Kubasik ◽  
G D Hallauer ◽  
R G Brodows
Keyword(s):  
Detection Limit ◽  
Luteal Phase ◽  
Solid Phase ◽  
Cross Reactivity ◽  
Luteal Function ◽  
Menstrual Cycles ◽  
Analytical Recovery ◽  
Solid Phase Radioimmunoassay ◽  
Clinically Significant ◽  
Direct Solid

Abstract We have evaluated a commercially available, direct, solid-phase radioimmunoassay kit for progesterone determination in serum or plasma. The assay is precise, within-run precision (CV) in the clinically significant ranges being 2.5 to 5.2%, between-run 5.5 to 5.8%. Mean analytical recovery of different concentrations of progesterone added to serum was 99.7% (range 95.3 to 102.7%). Fourteen closely related steroids showed no cross reactivity. The minimum detection limit was 0.5 microgram/L. Luteal-phase progesterone concentrations in serum were increased (greater than 3 micrograms/L) in 19 normal ovulatory menstrual cycles and decreased (less than 1.5 micrograms/L) in two nonovulatory cycles. We found this direct assay for progesterone to be analytically and clinically sound, and useful for assessing luteal-phase function.

Direct solid-phase enzymoimmunoassay of testosterone in saliva.

1989 ◽  
Vol 35 (10) ◽  
pp. 2044-2047 ◽  
Author(s):  
K Howard ◽  
M Kane ◽  
A Madden ◽  
J P Gosling ◽  
P F Fottrell
Keyword(s):  
Detection Limit ◽  
Analytical Procedure ◽  
Solid Phase ◽  
Medical Applications ◽  
Microtiter Plates ◽  
Large Numbers ◽  
Coefficients Of Variation ◽  
Serial Sampling ◽  
Direct Solid

Abstract This competitive, solid-phase enzymoimmunoassay for testosterone in saliva is carried out on microtiter plates and involves no chromatographic or extraction steps. With an overnight incubation the detection limit of the assay is 230 fg per well (16.1 pmol/L). There was a good correlation (correlation coefficient 0.95) between testosterone concentrations measured with and without prior extraction of the saliva samples. Repeated assay of three control saliva samples containing a range of testosterone concentrations (200-1000 pmol/L) gave within- and between-assay coefficients of variation of 5.5-13.2%. The analytical procedure is simple and closely resembles already published procedures for the determination of progesterone and estrone (with extraction) in saliva. One person can assay 200 samples in 24 h and the assay is suitable for reproductive and sports medical applications, particularly for projects involving serial sampling and yielding large numbers of samples.

Solid-phase enzymoimmunoassay for osteocalcin in human serum or plasma, with use of a monoclonal antibody.

1989 ◽  
Vol 35 (10) ◽  
pp. 2087-2092 ◽  
Author(s):  
M J Power ◽  
P F Fottrell
Keyword(s):  
Monoclonal Antibody ◽  
Detection Limit ◽  
Horseradish Peroxidase ◽  
Human Serum ◽  
Solid Phase ◽  
Sample Volume ◽  
Microtiter Plates ◽  
Analytical Recovery ◽  
Peroxidase Conjugate

Abstract In this solid-phase enzymoimmunoassay on microtiter plates for osteocalcin in serum or plasma, we use an osteocalcin-horseradish-peroxidase conjugate and a monoclonal antibody raised against bovine osteocalcin. We thoroughly standardized the assay for measurement of osteocalcin in both serum and plasma, demonstrating independence of sample volume, and determining the analytical recovery and within-and between-assay CVs. The detection limit was between 0.6 and 1.1 micrograms/L and the ED50 was 16 micrograms/L for a 5-microL sample volume. The intra-assay CV over the range 3 to 74 micrograms/L was less than or equal to 15%. The interassay CV over the range 3.6 to 46 micrograms/L was less than or equal to 16%. Results by this assay and by an in-house radioimmunoassay in which the same monoclonal antibody was used correlated well (r2 = 0.948). Osteocalcin concentrations in serum and plasma as measured with the present assay agreed well with published values.

Solid-phase enzymoimmunoassay of estrone in saliva.

1989 ◽  
Vol 35 (4) ◽  
pp. 569-572 ◽  
Author(s):  
J Folan ◽  
J P Gosling ◽  
M F Finn ◽  
P F Fottrell
Keyword(s):  
Detection Limit ◽  
Diethyl Ether ◽  
Analytical Procedure ◽  
Solid Phase ◽  
Ovarian Activity ◽  
Microtiter Plates ◽  
Coefficients Of Variation ◽  
Serial Sampling ◽  
High Concentrations ◽  
Working Day

Abstract In this solid-phase enzymoimmunoassay for estrone in saliva, microtiter plates are used after extraction of the sample with diethyl ether. No chromatographic step is involved. The detection limit of the assay is 450 fg per well (33 pmol/L). Intra- and interassay coefficients of variation for the assay of low, medium, and high concentrations of estrone in saliva were respectively 4.2, 12.7; 5.2, 8.7; and 2.7, 5.8%. Using this assay, we found a highly significant correlation (P less than 0.001) between estrone concentrations in time-matched serum and saliva samples. The analytical procedure is rapid and relatively simple. One person can assay 50-60 saliva samples during a normal working day. We conclude that the assay is very suitable, even in small laboratories, for saliva estrone measurements, which, in facilitating serial sampling, enables dynamic observations of estrone concentrations and ovarian activity to be more easily made.

Solid-phase enzymoimmunoassay of estrone in serum.

1988 ◽  
Vol 34 (9) ◽  
pp. 1843-1846 ◽  
Author(s):  
J Folan ◽  
J P Gosling ◽  
P F Fottrell
Keyword(s):  
Detection Limit ◽  
Correlation Coefficient ◽  
Diethyl Ether ◽  
Analytical Procedure ◽  
Solid Phase ◽  
Serum Samples ◽  
Microtiter Plates ◽  
Coefficients Of Variation ◽  
High Concentrations ◽  
Working Day

Abstract This rapid solid-phase enzymoimmunoassay for estrone in serum or plasma is done on microtiter plates after the serum is extracted with diethyl ether. No chromatographic or centrifugation steps are involved. The detection limit of the assay is 380 fg per well (28 pmol/L). Intra- and interassay coefficients of variation for the assay of low, medium, and high concentrations of estrone in plasma were respectively 4.4, 9.3; 2.3, 9.1; and 2.0, 6.3 percent. There was a good correlation (correlation coefficient 0.95) between estrone concentrations measured with this assay and with a commercial radioimmunoassay. The analytical procedure is simple, and one person can assay 80 serum samples per working day. We conclude that the assay is very suitable for serum estrone measurements and is more convenient than published radioimmunoassays.

Direct chemiluminescence immunoassay for estradiol in serum.

1986 ◽  
Vol 32 (10) ◽  
pp. 1895-1900 ◽  
Author(s):  
J De Boever ◽  
F Kohen ◽  
C Usanachitt ◽  
D Vandekerckhove ◽  
D Leyseele ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Solid Phase ◽  
Cross Reactivity ◽  
Serum Samples ◽  
Chemiluminescence Immunoassay ◽  
Binding Reaction ◽  
High Affinity ◽  
Microtiter Plates ◽  
Analytical Recovery

Abstract In this simple, reliable, fast solid-phase chemiluminescence immunoassay for directly measuring (i.e., without prior extraction) estradiol-17 beta in serum, a monoclonal antibody is used that binds estradiol with high affinity (Ka = 10(10) L/mol), and does not bind other steroids tested, the highest cross reactivity observed being 0.1% for estradiol-17 alpha. In this system the monoclonal antibody is bound to the wells of microtiter plates via a second antibody directed against the monoclonal antibody. Fifty microliters of serum and estradiol-displacing agents are added, followed by 100 pg of estradiol-isoluminol conjugate, and the label is measured by luminometry after the binding reaction. The sensitivity of the assay is 180 pmol per liter of serum, and the effective working range at less than or equal to 10% CV is 270 to 6700 pmol/L. Analytical recovery of added estradiol averaged 99.7% (SD 6.5%). Within- and between-assay CVs ranged between 5 and 12.7%. Thirty-five unknown serum samples can be assayed within 4 h. Results correlated well with those obtained with a direct RIA: r = 0.94 (n = 149). This assay opens new perspectives for chemiluminescence immunoassays.

Chemiluminescence immunoassay for somatomedin C in serum.

1987 ◽  
Vol 33 (11) ◽  
pp. 1989-1993 ◽  
Author(s):  
B Tode ◽  
G Messeri ◽  
F Bassi ◽  
M Pazzagli ◽  
M Serio
Keyword(s):  
Detection Limit ◽  
Solid Phase ◽  
Regression Equation ◽  
Sensitive Detection ◽  
Competitive Immunoassay ◽  
Chemiluminescence Immunoassay ◽  
Somatomedin C ◽  
Correlation Studies ◽  
Carboxy Terminal ◽  
Chemiluminescent Method

Abstract To select the best tracer for use in a competitive immunoassay, we conjugated human somatomedin C (SmC) to various chemiluminescent compounds via two different synthetic pathways. Naphthylhydrazides and arylhydrazides, used as the labels, were incorporated via their imidate or their succinimide esters. Conjugating the carboxy terminal of (amino ethyl)ethyl-isoluminol to SmC via a succinimide linkage supplied the most sensitive detection limit and the most immunoreactive conjugate. We developed an immunoassay based on the use of this conjugate, and evaluated dextran-coated charcoal, second-antibody precipitation, and solid-phase immunoprecipitation for separating bound and free label. This chemiluminescent method has a detection limit of 16 pg per tube, and it is accurate and precise. Correlation studies with a conventional radioimmunoassay (x) for SmC gave the following regression equation: y = 0.66x + 3.76 (r = 0.953, n = 30); the slight discrepancies between the two methods are probably ascribable to the use of different antibodies. We thus propose this chemiluminescence immunoassay as an inexpensive and sensitive alternative to radioimmunoassay for measuring SmC in serum or in extracts of serum.

Determination of Residual Acrylamide Monomer in Medical Polyacrylamide Hydrogel by HPLC-SPE

2005 ◽  
Vol 288-289 ◽  
pp. 405-408
Author(s):  
Zi Yi Wan ◽  
Ting Fei Xi ◽  
P. Zhao ◽  
Y. Sun ◽  
Z.G. Feng
Keyword(s):  
Detection Limit ◽  
Solid Phase ◽  
Polyacrylamide Hydrogel ◽  
Simple Method ◽  
Lower Detection Limit ◽  
Response Range ◽  
Coefficients Of Variation ◽  
Lower Detection ◽  
Linear Response Range

The polyacrylamide hydrogel (PAMG) has been used in cosmetology in China, Ukraine and Russia since 1990s. Because the monomer acrylamide(AM) used to produce PAMG has been implicated as a potential mutagen and reproductive toxicant[1,2], it is important to accurately determine the amount of residual AM monomer in the PAMG. In this study, a quick, practical and simple method to determine AM is presented with respect to the hydrogel. AM is analysed quantitatively by ODS-3 column with ultraviolet (UV) absorbance detector. AM is separated from interferential component with an aqueous solution of 0.9%NaCl (NS) adjusted at pH~3.7 using hydrochloric acid and then detected at a UV wavelength of 210 nm. The results show that ODS-3 is effective approach for quantifying AM concentrations in PAMG. This method has a lower detection limit of 0.003µg/ml and a linear response range of 0.003 and 0.9 µg/ml (depending on the range required for analysis). Precision studies give coefficients of variation of <3.2%(n=5) for 0.003µg/ml. The recoveries for this method are greater than 90%. When AM content in PAMG is lower than the detection limit of this method, SPE (solid phase extraction) could be used to concentrate AM. In the case, C18 cartridge is used. And the recoveries are about 70% for SPE when AM concentration is lower than ppb.

A fully automated solid-phase radioimmunoassay for triiodothyronine.

1977 ◽  
Vol 23 (5) ◽  
pp. 851-854 ◽  
Author(s):  
N E Rugg ◽  
M J Hasler ◽  
R E Bjornsen ◽  
K Painter
Keyword(s):  
Bovine Serum Albumin ◽  
Bovine Serum ◽  
Solid Phase ◽  
Correlation Coefficients ◽  
Ammonium Sulfate Precipitation ◽  
Men And Women ◽  
Coefficients Of Variation ◽  
Analytical Recovery ◽  
Solid Phase Radioimmunoassay ◽  
Normal Men

Abstract We report the first solid-phase radioimmunoassay for triiodothyronine with use of antibody-coated tubes, which is designed specifically for the fully automated radioimmunoassay instrument, Micromedic Systems' "Concept 4 Automatic Radioassay." Antisera to triiodothyropropionic acid/bovine serum albumin were raised in rabbits, purified by ammonium sulfate precipitation, and coated onto polypropylene tubes. Analytical recovery of exogenous triiodothyronine added to sera from normal men and women and pregnant women was quantitative. Intra-assay and inter-assay coefficients of variation are 4-5 and 6-9%, respectively. Correlation coefficients (r) for comparison of sample values with those obtained by three commercial laboratories were 0.95, 0.84, and 0.91. The sensitivity of the assay is 0.5 microng/liter. The assay can be performed either manually or be fully automated on the "Concept 4."

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