Plasma 21-deoxycortisol: comparison of a time-resolved fluoroimmunoassay using a biotinylated tracer with a radioimmunossay using 125iodine

Fiet, Jean; Boudi, Ahmed; Giton, Frank; Villette, Jean‐Marie; Boudou, P.; Soliman, Hany; Morineau, Gilles; Galons, Hervé

doi:10.1016/s0960-0760(99)00151-x

Cited by 17 publications

(10 citation statements)

References 13 publications

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“…The chromatographic separation of androgens and precursors, 4-androstenedione (A), 17-OHP, testosterone (T), 11-OHA, 21-DF, and the preparation and the qualities of antibodies prepared in-house have been previously reported [13]. Immunoassays were carried out using RIA for all these steroids, except for 21-DF, which was measured by means of sensitive, time-resolved fluoroimmunoassay [14]. Progesterone, 11-deoxycorticosterone (DOC), 21-DB, aldosterone, and cortisol were separated by Celite + ethylene glycol chromatography, using the following elution sequence: pure isooctane (5 ml), isooctane/dichloromethane (DCM) (88/12, v/v) 5 ml, isooctane/DCM (65/35, v/v) 5 ml, isooctane/DCM (20/80, v/v) 5 ml, isooctane/ethylacetate (20/80, v/v) 6 ml (unpublished data).…”

Section: Steroid Studiesmentioning

confidence: 99%

Testicular adrenal rest tumours in salt wasting congenital adrenal hyperplasia (in vivo and in vitro studies)

Bercovici

Fiet

Gibault

et al. 2005

The Journal of Steroid Biochemistry and Molecular Biology

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Section: Steroid Studiesmentioning

confidence: 99%

Testicular adrenal rest tumours in salt wasting congenital adrenal hyperplasia (in vivo and in vitro studies)

Bercovici

Fiet

Gibault

et al. 2005

The Journal of Steroid Biochemistry and Molecular Biology

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“…22,33 Europium chelates used as nonradioactive labels and their sensitive quantitation based on time-resolved fluorescence have been used in a wide variety of routine and research applications in human and veterinary medicine. 2,3,14,15,25,36,41,43 The europium chelates give a strong long decay time fluorescence, creating high specific activity in the labeled immunoreagents. 32 They are ideal labels for fluoroimmunoassays when high sensitivity is required.…”

Section: Introductionmentioning

confidence: 99%

C-Reactive Protein Measurement in Canine Saliva

et al. 2005

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Abstract. An established time-resolved immunofluorometric assay designed for measurement of C-reactive protein (CRP) in canine blood was evaluated and validated for use in canine saliva. C-reactive protein was measured in saliva specimens from 5 healthy dogs before and after the injection of casein and in 37 dogs with different disease conditions. The analytical and functional limits of detection were 0.000053 g/ml and 0.0091 g/ml, respectively, and intra-and interassay coefficients of variation ranged between 6.7-9.9% and 8.5-16.5%, respectively. A recovery experiment showed no significant disagreement between detected values and expected ones, and saliva CRP concentration was measured in a linear and proportional manner. A positive correlation was found between CRP levels obtained in saliva and serum samples in the experimental (R 2 ϭ 0.76) and clinical studies (R 2 ϭ 0.70). The assay was able to detect significant differences between salivary CRP levels in healthy dogs and dogs with inflammatory processes. These results suggest that saliva can be used for CRP measurement in dogs. The use of saliva presents the advantage of an easier and less stressful sampling method for the animals, which might be performed outside of hospital environments.

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“…Time-resolved fluoroimmunoassays are a non-radioactive label technology which offers rapid and ultrasensitive tests for the measurement of several animal and human hormones [11,12,25,33] and also for the detection of drugs [15], specific antibodies [1,27] and cancer markers [32,37]. Other possible applications are the determination of acute phase proteins such as bovine haptoglobin [21] or human and canine C-reactive protein [26,34].…”

Section: Introductionmentioning

confidence: 99%

Development of a time-resolved fluorometry based immunoassay for the determination of canine haptoglobin in various body fluids

Parra

Väisänen

Cerón³

2005

Vet. Res.

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-A time-resolved immunofluorometric assay (TR-IFMA) was developed for the determination of haptoglobin (Hp) in canine serum. Haptoglobin was purified from canine acute phase serum by ammonium sulphate precipitation followed by gel filtration. This isolated dog Hp was used as the standard to calibrate the assay. Intra-and inter-assay coefficients of variation of the assay were, respectively, 5.7% and 16.6% at 0.51 mg/mL, 2.4% and 10.6% at 2.1 mg/mL and 10.5% and 11.9% at 32.5 mg/mL. The dilution of serum samples with high Hp concentrations resulted in linear regression equations with R 2 of 0.99 and 0.97. A high correlation was found in serum Hp measurements by TR-IFMA and a commercial assay based on peroxidase activity of haemoglobin bound to haptoglobin (R 2 = 0.96). The limit of detection for the TR-IFMA method was 0.002 µg/mL. The addition of fresh haemolysate to serum samples did not affect the haptoglobin concentration (P = 0.694). Statistical differences (P < 0.003) were found between healthy dogs and dogs with different pathological processes. In whole blood, Hp concentrations were much lower than in serum but closely related (R 2 = 0.84) whereas saliva Hp concentrations were poorly related with serum concentrations (R 2 = 0.53). However, the concentration of Hp in saliva was significantly (P < 0.039) higher in dogs with pathological processes compared to healthy dogs. The assay sensitivity was adequate to also be applied to whole blood and saliva specimens.canine / haptoglobin / time-resolved fluorometry / whole blood / saliva

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Plasma 21-deoxycortisol: comparison of a time-resolved fluoroimmunoassay using a biotinylated tracer with a radioimmunossay using 125iodine

Cited by 17 publications

References 13 publications

Testicular adrenal rest tumours in salt wasting congenital adrenal hyperplasia (in vivo and in vitro studies)

Testicular adrenal rest tumours in salt wasting congenital adrenal hyperplasia (in vivo and in vitro studies)

C-Reactive Protein Measurement in Canine Saliva

Development of a time-resolved fluorometry based immunoassay for the determination of canine haptoglobin in various body fluids

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