Microfilter Paper Method for 17α-Hydroxyprogesterone Radioimmunoassay: Its Application for Rapid Screening for Congenital Adrenal Hyperplasia

Pang, Songja; Hotchkiss, J.; Drash, Allan L.; Levine, Lenore S.; New, M.I.

doi:10.1210/jcem-45-5-1003

Cited by 263 publications

(84 citation statements)

References 11 publications

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“…Elevated 17-OHP blood level is normally used as an indicator of CAH. In general, the technique of analyzing 17-OHP in neonatal screenings is radioimmunoassay in fi lter-paper blood samples (6,7). It has been discussed by several authors that this procedure generate a high rate of positive results attributable to physiological reasons and to cross-reactions with steroids other than 17-OHP, especially in preterm neonates (9,10).…”

Section: Discussionmentioning

confidence: 99%

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Heterozygosis for CYP21A2 mutation considered as 21-hydroxylase deficiency in neonatal screening

Soardi

Lemos-Marini

Coeli

et al. 2008

Arq Bras Endocrinol Metab

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Steroid 21-hydroxylase defi ciency (21-OHD) accounts for more than 90% of congenital adrenal hyperplasia. CAH newborn screening, in general, is based on 17-hydroxyprogesterone dosage (17-OHP), however it is complicated by the fact that healthy preterm infants have high levels of 17-OHP resulting in false positive cases. We report on molecular features of a boy born pre-term (GA = 30 weeks; weight = 1,390 g) with elevated levels of 17-OHP (91.2 nmol/L, normal < 40) upon neonatal screening who was treated as having CAH up to the age of 8 months. He was brought to us for molecular diagnosis. Medication was gradually suspended and serum 17-OHP dosages mantained normal. The p.V281L mutation was found in compound heterozygous status with a group of nucleotide alterations located at the 3` end intron 4 and 5' end exon 5 corresponding to the splice site acceptor region. Molecular studies continued in order to exclude the possibility of a nonclassical 21-OHD form. The group of three nucleotide changes was demonstrated to be a normal variant since they failed to interfere with the normal splicing process upon minigene studies. A defi ciência de 21-hidroxilase (21-OHD) é uma doença autossômica recessiva que contribui com mais de 90% dos casos de hiperplasia congênita da adrenal. O teste de dosagem de 17-hidroxiprogesterona (17-OHP) por radioimunoensaio em amostras de sangue colhidas em papel de fi ltro tem sido o método mais usado nos programas de triagem neonatal. No entanto, essa triagem pode apresentar alto número de falso-positivos pelo fato de os recém-nascidos prematuros apresentarem dosagens mais elevadas deste esteróide. Apresentamos aqui os estudos moleculares de uma criança, sexo masculino, nascida pré-termo (IG = 30 sem; peso = 1.390 g) que apresentava valores elevados de 17-OHP sérica (91,2 nmol/L, normal < 40) na triagem neonatal e que foi tratada como portadora da forma clássica da 21-OHD até a idade de 8 meses quando nos foi encaminhada para diagnóstico molecular. A terapia foi, então, gradativamente descontinuada, sendo que as concentrações séricas de 17-OHP se mantiveram normais. A mutação p.V281L foi encontrada em heterozigose composta com um grupo de alterações no terminal 3' do íntron 4 e no terminal 5' do éxon 5 correspondendo à região do sítio aceptor de splicing. A análise do gene CYP21A2 prosseguiu para se excluir a possibilidade de a criança ser afetada com a forma não-clássica de 21-OHD. Pela análise de minigene fi cou demonstrado que o grupo de três trocas nucleotídicas não afeta o processo normal de transcrição. Concluindo, a criança é apenas heterozigota da mutação p.V281L sem necessidade de tratamento.

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Section: Discussionmentioning

confidence: 99%

“…There is a high number of different techniques for diagnosing 21-OHD, but most CAH newborn screening is based on 17-hydroxyprogesterone radioimmunoassay on dried blood on fi lter paper (6)(7)(8). Nationwide and regional CAH screening programs have been introduced in several countries (7,9,10) including in a few regions in Brazil (11,12).…”

Section: Introductionmentioning

confidence: 99%

Heterozygosis for CYP21A2 mutation considered as 21-hydroxylase deficiency in neonatal screening

Soardi

Lemos-Marini

Coeli

et al. 2008

Arq Bras Endocrinol Metab

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“…17-Hydroxyprogesterone (17-OHP) is one of the biomarker molecules for the early diagnosis of CAH. Following the development of an immunoassay to determine the 17-OHP concentration in blood spots in 1977 (Pang et al, 1977), population newborn screening was introduced. Unfortunately, due to a physiologically delayed expression of the enzyme 11β-hydroxylase in premature babies, illness, birth stress, impaired kidney function, interference from 17-OHP metabolite(s) (Fingerhut, 2009) and cross-reactivity of antibodies with other steroids (Wong et al, 1992), particularly 17-hydroxypregnenolone (17-OHPreg), the positive predictive rate for CAH population screening using immunoassay remains below 1%, despite attempts to adjust cutoff levels for birth weight, gestational age and stress factors (Matern et al, 2007).…”

Section: Applications Newborn Screeningmentioning

confidence: 99%

Dried blood spot sampling in combination with LC‐MS/MS for quantitative analysis of small molecules

Li¹,

Tse²

2009

Biomedical Chromatography

536

428

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The collection of whole blood samples on paper, known as dried blood spot (DBS), dates back to the early 1960s in newborn screening for inherited metabolic disorders. DBS off ers a number of advantages over conventional blood collection. As a less invasive sampling method, DBS off ers simpler sample collection and storage and easier transfer, with reduced infection risk of various pathogens, and requires a smaller blood volume. To date, DBS-LC-MS/MS has emerged as an important method for quantitative analysis of small molecules. Despite the increasing popularity of DBS-LC-MS/MS, the method has its limitations in assay sensitivity due to the small sample size. Sample quality is often a concern. Systematic assessment on the potential impact of various blood sample properties on accurate quantifi cation of analyte of interest is necessary. Whereas most analytes may be stable on DBS, unstable compounds present another challenge for DBS as enzyme inhibitors cannot be conveniently mixed during sample collection. Improvements on the chemistry of DBS card are desirable. In addition to capturing many representative DBS-LS-MS/MS applications, this review highlights some important aspects of developing and validating a rugged DBS-LC-MS/MS method for quantitative analysis of small molecules along with DBS sample collection, processing and storage.

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“…This technique was initially developed in Alaska and used radioimmunoassay developed by Pang and cols. (12).…”

mentioning

confidence: 99%

Ten-year evaluation of a Neonatal Screening Program for Congenital Adrenal Hyperplasia

Nascimento

Cristiano

Campos

et al. 2014

Arq Bras Endocrinol Metab

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Objective: Evaluate the Neonatal Screening Program (NSP) for congenital adrenal hyperplasia (CAH) of the Department of Health of the State of Santa Catarina (Secretaria de Estado da Saúde de Santa Catarina, SES/SC), and provide information to improve the program. Subjects and methods: Descriptive, retrospective study of 748,395 children screened between January 2001 and December 2010. We analyzed the coverage of the NSP-SES/SC prevalence of CAH, child's age when the first sample for 17-hydroxyprogesterone (17OHP) measurement was collected, levels of 17OHP, mean age at treatment onset and main clinical manifestations. Results: The NSP-SES/ SC covered 89% of the live newborns in the State. It diagnosed 50 cases of CAH, yielding an incidence of 1:14,967. Mean age at collection of the first sample was 7.3 days and mean level of 17OHP was 152.9 ng/mL. The most frequent manifestations were virilized genitalia with nonpalpable gonads, clitoromegaly and genital hyperpigmentation. In three girls, the genre established at birth was incorrect. The salt-wasting form was present in 74% of the cases. There was no occurrence of shock or death. Mean age at treatment onset in the salt-wasting form was 17.4 days compared with 54.9 days in those without the salt-wasting form of the disease. All children were treated with hydrocortisone, and those with salt-wasting CAH were also treated with fludrocortisone. Conclusions: The incidence of CAH was 1 case to 14,967 live newborns. Collection of the first sample occurred outside the recommended time, resulting in delays in treatment onset. Arq Bras Endocrinol Metab. 2014;58(7):765-71

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Microfilter Paper Method for 17α-Hydroxyprogesterone Radioimmunoassay: Its Application for Rapid Screening for Congenital Adrenal Hyperplasia

Cited by 263 publications

References 11 publications

Heterozygosis for CYP21A2 mutation considered as 21-hydroxylase deficiency in neonatal screening

Heterozygosis for CYP21A2 mutation considered as 21-hydroxylase deficiency in neonatal screening

Dried blood spot sampling in combination with LC‐MS/MS for quantitative analysis of small molecules

Ten-year evaluation of a Neonatal Screening Program for Congenital Adrenal Hyperplasia

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