8c2dd974172ded4eb63632284a7211e7 pmc_33809035.pdf d962a5d9d5c0452c3d44389371296c50faa1f714 pmc_33809035.pdf a353a68327edaa61b632dc72f02657d0ccc5eaba9bc67da5c17990bb2e1b3125 pmc_33809035.pdf Title: Comprehensive Genetic Testing of CYP21A2: A Retrospective Analysis in Patients with Suspected Congenital Adrenal Hyperplasia Subject: The most common form of congenital adrenal hyperplasia (CAH) results from a deficiency of the 21-hydroxylase enzyme (21-OHD), presenting with a broad spectrum of clinical phenotypes according to the CYP21A2 gene mutations. Of the 59 patients with suspected CAH, 62.7% presented a positive genetic result. Of them, 78.4% and 18.9% presented with non-classical and classical forms, respectively. An overall phenotype-genotype correlation of 88.9% was observed. Biochemically, 17-hydroxiprogesterone concentrations were significantly higher in genetically confirmed patients. Genetically, 36 patients presented with previously reported pathogenic variants, and one presented a new variant in homozygosis. Among the 74 alleles tested, point mutations were found in 89.2% and large rearrangements were found in the rest. The most prevalent pathogenic variant was p.(Val282Leu). The inclusion of relatives revealed one further case. Interestingly, 87.5% of relatives were carriers of a pathogenic variant, including two siblings initially classified as genetically positive. In addition, the study of male partners with gestational desire identified several carriers of mild mutations. Studying the allelic distribution of the variants also allowed for reclassifying one patient. In conclusion, a genetic approach including Sanger sequencing, multiplex ligation-dependent probe amplification (MLPA) analysis, and allelic distribution of the pathogenic variants represents a beneficial tool for better classifying patients with 21-OHD. Keywords: congenital adrenal hyperplasia; CYP21A2 gene; genotype analysis; 17-OH progesterone; genotype-phenotype correlation Author: Madalina Nicoleta Nan, Rosa Roig, Susana Martínez, Jose Rives, Eulàlia Urgell, Juan José Espinós, Mireia Tirado, Gemma Carreras, Anna Aulinas, Susan M. Webb, Rosa Corcoy, Francisco Blanco-Vaca and Mireia Tondo Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Tue Mar 16 16:23:09 2021 CET ModDate: Tue Mar 16 16:27:46 2021 CET Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 13 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 481217 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- EMTEKM+VnURWPalladioL Type 1 Custom yes yes no 10 0 PAFJWR+URWPalladioL-Ital Type 1 Custom yes yes no 15 0 XLMNKV+URWPalladioL-Bold Type 1 Custom yes yes no 20 0 MYMRJC+URWPalladioL-BoldItal Type 1 Custom yes yes no 24 0 UNXUVA+URWPalladioL-Roma Type 1 Custom yes yes no 28 0 ZSBPMM+VnURWPalladioL-Bold Type 1 Custom yes yes no 32 0 XPUSBJ+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 40 0 BSDWLC+TimesNewRomanPSMT Type 1C WinAnsi yes yes no 43 0 MCBCCI+SourceSansRoman-Regular Type 1C Custom yes yes no 65 0 MCBCCM+SourceSansRoman.613wght Type 1C Custom yes yes no 69 0 MCBCCO+SourceSansRoman-Bold Type 1C Custom yes yes no 73 0 HIJFVP+CMSY10 Type 1 Builtin yes yes no 88 0 JFDKCD+Calibri CID TrueType Identity-H yes yes yes 109 0 JFDHKK+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 115 0 JFDHNL+PalatinoLinotype TrueType WinAnsi yes yes no 118 0 JFDHNN+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 121 0 JFDJOD+TimesNewRoman,Bold CID TrueType Identity-H yes yes yes 124 0 QDCWHF+EURM10 Type 1 Builtin yes yes no 140 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-10-02 03:24:12 CEST RepresentationInformation: pmc_33809035.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2023-03-31 12:09:34 CEST Size: 481217 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 311 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Materials and Methods Destination: section.2 Children: Item: Title: Study Participants Destination: subsection.2.1 Item: Title: Ethical Aspects Destination: subsection.2.2 Item: Title: Biochemical Parameters Destination: subsection.2.3 Item: Title: Genetic Screening Destination: subsection.2.4 Item: Title: Characterization of Variants and Bioinformatics Analysis Destination: subsection.2.5 Item: Title: Statistical Analysis Destination: subsection.2.6 Item: Title: Results Destination: section.3 Children: Item: Title: Study Cohort Characteristics Destination: subsection.3.1 Item: Title: Genetic Results Destination: subsection.3.2 Item: Title: Genetic Studies of Relatives Destination: subsection.3.3 Item: Title: Allelic Distribution of the Point Mutation Pathological Variants Destination: subsection.3.4 Item: Title: Discussion Destination: section.4 Item: Title: Conclusions Destination: section.5 Item: Title: References Destination: section.5 Info: Title: Comprehensive Genetic Testing of CYP21A2: A Retrospective Analysis in Patients with Suspected Congenital Adrenal Hyperplasia Author: Madalina Nicoleta Nan, Rosa Roig, Susana Martínez, Jose Rives, Eulàlia Urgell, Juan José Espinós, Mireia Tirado, Gemma Carreras, Anna Aulinas, Susan M. Webb, Rosa Corcoy, Francisco Blanco-Vaca and Mireia Tondo Subject: The most common form of congenital adrenal hyperplasia (CAH) results from a deficiency of the 21-hydroxylase enzyme (21-OHD), presenting with a broad spectrum of clinical phenotypes according to the CYP21A2 gene mutations. Of the 59 patients with suspected CAH, 62.7% presented a positive genetic result. Of them, 78.4% and 18.9% presented with non-classical and classical forms, respectively. An overall phenotype-genotype correlation of 88.9% was observed. Biochemically, 17-hydroxiprogesterone concentrations were significantly higher in genetically confirmed patients. Genetically, 36 patients presented with previously reported pathogenic variants, and one presented a new variant in homozygosis. Among the 74 alleles tested, point mutations were found in 89.2% and large rearrangements were found in the rest. The most prevalent pathogenic variant was p.(Val282Leu). The inclusion of relatives revealed one further case. Interestingly, 87.5% of relatives were carriers of a pathogenic variant, including two siblings initially classified as genetically positive. In addition, the study of male partners with gestational desire identified several carriers of mild mutations. Studying the allelic distribution of the variants also allowed for reclassifying one patient. In conclusion, a genetic approach including Sanger sequencing, multiplex ligation-dependent probe amplification (MLPA) analysis, and allelic distribution of the pathogenic variants represents a beneficial tool for better classifying patients with 21-OHD. 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