{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T16:59:44Z","timestamp":1780505984352,"version":"3.54.1"},"reference-count":22,"publisher":"Public Library of Science (PLoS)","issue":"5","license":[{"start":{"date-parts":[[2021,5,20]],"date-time":"2021-05-20T00:00:00Z","timestamp":1621468800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Hungarian-Romanian Cross Border Cooperation Project","award":["0802\/008 SCREENGEN"],"award-info":[{"award-number":["0802\/008 SCREENGEN"]}]},{"DOI":"10.13039\/501100015763","name":"University of Szeged","doi-asserted-by":"crossref","award":["Open Access Fund 5121"],"award-info":[{"award-number":["Open Access Fund 5121"]}],"id":[{"id":"10.13039\/501100015763","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>In newborn screening, samples suspected for congenital adrenal hyperplasia (CAH), a potentially lethal inborn error of steroid biosynthesis, need to be confirmed using liquid chromatography\u2013tandem mass spectrometry. Daily quality controls (QCs) for the 2<jats:sup>nd<\/jats:sup>-tier CAH assay are not commercially available and are therefore generally prepared within the laboratory. For the first time, we aimed to compare five different QC preparation approaches used in routine diagnostics for CAH on the concentrations of cortisol, 21-deoxycortisol, 11-deoxycortisol, 4-androstenedione and 17-hydroxyprogesterone in dried blood spots. The techniques from Prep1 to Prep5 were tested at two analyte concentrations by spiking aliquots of a steroid-depleted blood, derived from washed erythrocyte suspension and steroid-depleted serum. The preparation processes differed in the sequence of the preparation steps and whether freeze-thaw cycles were used to facilitate blood homogeneity. The five types of dried blood spot QCs were assayed and quantitated in duplicate on five different days using a single calibration row per day. Inter-assay variations less than 15% and concentrations within \u00b115% of the nominal values were considered acceptable. Results obtained by means of the four dried blood spot QC preparation techniques (Prep1, Prep2, Prep4 and Prep5) were statistically similar and remained within the \u00b115% ranges in terms of both reproducibility and nominal values. However, concentration results for Prep3 (spiking prior to three freeze-thaw cycles) were significantly lower than the nominal values in this setting, with differences exceeding the \u00b115% range in many cases despite acceptable inter-assay variations. These findings have implications for the in-house preparation of QC samples in laboratory developed tests for CAH, including 2<jats:sup>nd<\/jats:sup>-tier assays in newborn screening.<\/jats:p>","DOI":"10.1371\/journal.pone.0252091","type":"journal-article","created":{"date-parts":[[2021,5,20]],"date-time":"2021-05-20T17:57:30Z","timestamp":1621533450000},"page":"e0252091","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":6,"title":["Comparison of different preparation techniques of dried blood spot quality controls in newborn screening for congenital adrenal hyperplasia"],"prefix":"10.1371","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3355-9113","authenticated-orcid":true,"given":"N\u00f3ra","family":"Grecs\u00f3","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anita","family":"Z\u00e1dori","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"\u00c1kos","family":"Bar\u00e1th","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zsolt","family":"Galla","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"G\u00e1bor","family":"R\u00e1cz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Csaba","family":"Bereczki","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3591-6054","authenticated-orcid":true,"given":"P\u00e9ter","family":"Monostori","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2021,5,20]]},"reference":[{"key":"pone.0252091.ref001","doi-asserted-by":"crossref","first-page":"2194","DOI":"10.1016\/S0140-6736(17)31431-9","article-title":"Congenital adrenal hyperplasia","volume":"390","author":"D El-Maouche","year":"2017","journal-title":"Lancet"},{"key":"pone.0252091.ref002","doi-asserted-by":"crossref","first-page":"490","DOI":"10.1038\/nrendo.2009.148","article-title":"Neonatal screening for congenital adrenal hyperplasia","volume":"5","author":"PC White","year":"2009","journal-title":"Nat Rev Endocrinol"},{"key":"pone.0252091.ref003","doi-asserted-by":"crossref","first-page":"15","DOI":"10.3390\/ijns7010015","article-title":"Neonatal Screening in Europe Revisited: An ISNS Perspective on the Current State and Developments Since 2010","volume":"7","author":"JG Loeber","year":"2021","journal-title":"Int J Neonatal Screen"},{"key":"pone.0252091.ref004","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1016\/j.mce.2012.12.026","article-title":"Mass spectrometry theory and application to adrenal diseases","volume":"371","author":"KM Wooding","year":"2013","journal-title":"Mol Cell Endocrinol"},{"key":"pone.0252091.ref005","doi-asserted-by":"crossref","first-page":"263","DOI":"10.3343\/alm.2019.39.3.263","article-title":"Dried Blood Spot Multiplexed Steroid Profiling Using Liquid Chromatography Tandem Mass Spectrometry in Korean Neonates","volume":"39","author":"R Choi","year":"2019","journal-title":"Ann Lab Med"},{"key":"pone.0252091.ref006","doi-asserted-by":"crossref","first-page":"375","DOI":"10.4155\/bio.16.6","article-title":"Determination of a steroid profile in heel prick blood using LC\u2013MS\/MS","volume":"8","author":"A Boelen","year":"2016","journal-title":"Bioanalysis"},{"key":"pone.0252091.ref007","doi-asserted-by":"crossref","first-page":"311","DOI":"10.1159\/000439380","article-title":"Concurrent Confirmation and Differential Diagnosis of Congenital Adrenal Hyperplasia from Dried Blood Spots: Application of a Second-Tier LC-MS\/MS Assay in a Cross-Border Cooperation for Newborn Screening","volume":"84","author":"P Monostori","year":"2015","journal-title":"Horm Res Paediatr"},{"key":"pone.0252091.ref008","doi-asserted-by":"crossref","first-page":"1437","DOI":"10.1016\/j.steroids.2011.07.013","article-title":"Rapid steroid hormone quantification for congenital adrenal hyperplasia (CAH) in dried blood spots using UPLC liquid chromatography-tandem mass spectrometry","volume":"76","author":"N Janzen","year":"2011","journal-title":"Steroids"},{"key":"pone.0252091.ref009","doi-asserted-by":"crossref","first-page":"621","DOI":"10.1373\/clinchem.2003.027193","article-title":"Improved specificity of newborn screening for congenital adrenal hyperplasia by second-tier steroid profiling using tandem mass spectrometry","volume":"50","author":"JM Lacey","year":"2004","journal-title":"Clin Chem"},{"key":"pone.0252091.ref010","unstructured":"CDC 2020 Set1 Quality Control Report. 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