Plasma 17-hydroxyprogesterone determination with two commercial immunoassays

Nahoul, K

doi:10.1016/0960-0760(94)90029-9

Cited by 11 publications

(8 citation statements)

References 16 publications

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“…Screening for CAH in newborns has become a routine part of many programs by measuring levels of 17αhydroxyprogesterone (17OH-P) (1,2). Currently, neonatal screening procedures for CAH are based on immunoassays (3,4). This approach, while practical, lacks some degree of specificity because of cross-reacting congeners that cannot be separated from 17OH-P in the direct assay (4)(5)(6).…”

Section: Introductionmentioning

confidence: 99%

Rapid screening assay of congenital adrenal hyperplasia by measuring 17α‐hydroxyprogesterone with high‐performance liquid chromatography/electrospray ionization tandem mass spectrometry from dried blood spots

Lai

Tsai

et al. 2002

Clinical Laboratory Analysis

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A rapid, simple, and specific method was developed for the diagnosis of congenital adrenal hyperplasia (CAH) from dried blood spots on newborn screening cards based on high-performance liquid chromatography/electrospray ionization tandem mass spectrometry (HPLC/ESI-MS/MS). The usefulness of 17 alpha-hydroxyprogesterone (17 OH-P) determination on dried filter-paper blood samples from patients with CAH caused by 21-hydroxylase deficiency was evaluated. The LC/MS/MS detection of 17 OH-P was rapid, <4 min. The intra- and interday accuracy and precision of the method were <7%. Our procedure maintained good linearities (R(2) > 0.992) and recovery rate (>83%). We used this new method to directly determine the 17 OH-P levels in dried blood specimens from abnormal children of various ages, with a detection limit of 20 ng/ml (approximately 240 pg), to avoid the time-consuming derivatization steps required by the gas-chromatography/mass spectrometry (GC/MS) method. Four dried filter-paper blood samples of CAH patients (three girls and one boy, 1-14 years old) were all quantified in an LC/MS/MS study and revealed high 17 OH-P levels (>90 ng/ml). After treatment, all of the elevated 17 OH-P levels either decreased or disappeared. Compared with CAH patients, 17 OH-P was nearly undetectable (<20 ng/ml) in the normal infants by LC/MS/MS. This LC/MS/MS assay is not only useful for both diagnosis and monitoring of treatment of CAH in all other age groups, it also can be used as a screening test for CAH infants. In this study, we provided the first data on 17 OH-P in dried blood specimens affected with CAH using HPLC/ESI-MS/MS.

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Section: Introductionmentioning

confidence: 99%

Rapid screening assay of congenital adrenal hyperplasia by measuring 17α‐hydroxyprogesterone with high‐performance liquid chromatography/electrospray ionization tandem mass spectrometry from dried blood spots

Lai

Tsai

et al. 2002

Clinical Laboratory Analysis

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“…Monitoring and screening for CAH patients by measuring levels of 17α‐hydroxyprogesterone (17OHP) or other steroids has become a routine part of many programmes 1,. 2 Numerous methods have been described to determine these steroid hormones, such as fluorimetry,3,, 4 radioimmunoassay (RIA),5–8 and high‐performance liquid chromatography (HPLC) 4,. 9,, 10 Most of these, however, are affected by a degree of interference or cross‐reactivity with other steroids.…”

mentioning

confidence: 99%

Rapid monitoring assay of congenital adrenal hyperplasia with microbore high‐performance liquid chromatography/electrospray ionization tandem mass spectrometry from dried blood spots

Lai

Tsai

et al. 2001

Rapid Comm Mass Spectrometry

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17-hydroxyprogesterone (17OHP) is the most important plasma parameter for diagnosing and monitoring congenital adrenal hyperplasia (CAH) caused by 21-hydroxylase deficiency. A rapid, simple, and specific method based on microbore high-performance liquid chromatography/electrospray ionization tandem mass spectrometry (micro-HPLC/ESI-MS/MS) was developed to determine the presence of 17OHP on dried filter-paper blood samples from patients with CAH caused by 21-hydroxylase deficiency. 17OHP from dried blood spots formed by the action of Girard reagent P (GirP) turned out to be a water-soluble hydrazone complex. Derivatization with GirP led to higher ESI sensitivity for 17OHP. The LC/MS/MS detection of GirP-derivatized 17OHP (GirP-17OHP) was rapid (<3 min). The method is repeatable and reproducible, with CVs <7% and 12%, respectively. This new method was used for direct determination of 17OHP in dried blood specimens obtained from abnormal children and infants of various ages with a detection limit of 10 ng/mL ( approximately 12 microL blood). The method described allows for rapid and reliable measurements of 17OHP in dried blood specimens from patients affected by CAH.

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“…Moreover, high 17-OHPreg levels may also interfere with the most commonly used laboratory methods (such as RIA protocols) to measure 17-OHP levels [20,21]. In our study, 17-OHP was measured by an enzyme immunoassay (17α-OH-progesterone Elisa, DRG Instruments GmbH) and cross reactivity with 17-OHPreg was not tested because the assay for 17-OHPreg was not available in our laboratory.…”

Section: Discussionmentioning

confidence: 99%

A New Homozygous Frameshift Mutation in the <b><i>HSD3B2</i></b> Gene in an Apparently Nonconsanguineous Italian Family

et al. 2016

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Background: 3β-Hydroxysteroid dehydrogenase (3β-HSD) deficiency is a rare cause of congenital adrenal hyperplasia (CAH) caused by inactivating mutations in the HSD3B2 gene. Patient and Methods: We report the molecular and structural analysis of the HSD3B2 gene in a 46,XY child born to apparently nonconsanguineous parents and presenting ambiguous genitalia and salt wasting. The steroid profile showed elevated concentrations of 17-hydroxyprogesterone, androstenedione, ACTH and plasma renin, but normal values of cortisol and dehydroepiandrosterone sulfate. Unexpectedly, plasma aldosterone was high. For structural and functional analyses, the three-dimensional structure of 3β-HSD2 was modeled using the crystal structure of the short-chain dehydrogenase Gox2253 from Gluconobacter oxydans as a template. Results: The direct DNA sequence of the child revealed a new homozygous frameshift mutation in exon 4 of the HSD3B2 gene, a single nucleotide deletion at codon 319 [GTC(Val)→GC], yielding premature stop codon in position 367. Molecular homology modeling and secondary structure predictions suggested that the variant sequence might both alter the substrate-binding cleft and compromise the overall stability of the enzyme. Conclusion: We have described the first HSD3B2 gene mutation in the Italian population and analyzed its effect in the context of the 3β-HSD2 structure and function.

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Plasma 17-hydroxyprogesterone determination with two commercial immunoassays

Cited by 11 publications

References 16 publications

Rapid screening assay of congenital adrenal hyperplasia by measuring 17α‐hydroxyprogesterone with high‐performance liquid chromatography/electrospray ionization tandem mass spectrometry from dried blood spots

Rapid screening assay of congenital adrenal hyperplasia by measuring 17α‐hydroxyprogesterone with high‐performance liquid chromatography/electrospray ionization tandem mass spectrometry from dried blood spots

Rapid monitoring assay of congenital adrenal hyperplasia with microbore high‐performance liquid chromatography/electrospray ionization tandem mass spectrometry from dried blood spots

A New Homozygous Frameshift Mutation in the <b><i>HSD3B2</i></b> Gene in an Apparently Nonconsanguineous Italian Family

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