8c2dd974172ded4eb63632284a7211e7 255430.pdf d962a5d9d5c0452c3d44389371296c50faa1f714 255430.pdf a353a68327edaa61b632dc72f02657d0ccc5eaba9bc67da5c17990bb2e1b3125 255430.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 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. 1.22.1, 2019-04-17) Date: 2022-03-10 03:03:13 CET RepresentationInformation: 255430.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2022-03-09 17:20:20 CET 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: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7b69c6ba Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@46daef40 Children: Item: Title: Study Participants Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@12f41634 Item: Title: Ethical Aspects Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@13c27452 Item: Title: Biochemical Parameters Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Item: Title: Genetic Screening Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Item: Title: Characterization of Variants and Bioinformatics Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5ed828d Item: Title: Statistical Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@50d0686 Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a3d45bd Children: Item: Title: Study Cohort Characteristics Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1e7c7811 Item: Title: Genetic Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: Genetic Studies of Relatives Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Item: Title: Allelic Distribution of the Point Mutation Pathological Variants Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@a38d7a3 Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77f99a05 Item: Title: Conclusions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@63440df3 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@3aeaafa6 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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