- Is a 49 gene panel that includes assessment of non-coding variants
Is ideal for patients presenting with ambiguous genitalia, patients suspected to have a disorder of sexual development and patients suspected to have congenital adrenal hyperplasia (CAH).
Number of genes49
CPT codesSEQ 81405
The Blueprint Genetics Abnormal Genitalia/ Disorders of Sex Development Panel (test code EN0201):
- Is a 49 gene panel that includes assessment of selected non-coding disease-causing variants
- Is available as PLUS analysis (sequencing analysis and deletion/duplication analysis), sequencing analysis only or deletion/duplication analysis only
Commonly used ICD-10 code(s) when ordering the Abnormal Genitalia/ Disorders of Sex Development Panel
|Q56.0||Indeterminate sex and pseudohermaphroditism|
|E34.50||Androgen insensitivity syndrome|
|E25.0||Congenital adrenal hyperplasia|
- EDTA blood, min. 1 ml
- Purified DNA, min. 3μg
- Saliva (Oragene DNA OG-500 kit)
Label the sample tube with your patient’s name, date of birth and the date of sample collection.
Note that we do not accept DNA samples isolated from formalin-fixed paraffin-embedded (FFPE) tissue.
Disorders of sex development (DSD) are a group of congenital conditions characterized by problems in the course of gender patterning, gonadal and sex development. It has been estimated that 1% – 2% of live births have some aspect of DSD. Approximately 5% of infants with DSD have ambiguous genitalia and indeterminate sex at birth. However, the vast majority of these patients do not require corrective surgery. Patients with 46,XY DSD have often impaired androgen synthesis or action and may have normal female external genitalia, while patients with 46,XX DSD conditions have often androgen excess. In 46,XX females, congenital adrenal hyperplasia (CAH) caused by 21-hydroxylase deficiency (21-OHD) is the most common cause of DSD. The estimated prevalence of CAH is 1:10,000 and 90%-95% of cases are due to mutations in CYP21A2. The severity of the condition often depends on the residual enzyme activity subdiving CYP21A2 mutations in severe (classic phenotype, enzyme activity 0%-10%) and mild (non-classic, enzyme activity 20%-50%). Androgen insensitivity syndrome (AIS), caused by mutations in AR, is characterized by feminization of external genitalia and atypical sexual development in 46,XY individuals. The condition may be complete, partial or mild, depending on the level of androgen insensitivity. Mutations in the AR gene explain up to 95% of cases with complete androgen insensitivity, while the proportions are lower for the partial and mild subtypes. The combined prevalence of various AIS subtypes is estimated to be 5:100,000.
Genes in the Abnormal Genitalia/ Disorders of Sex Development Panel and their clinical significance
|AMH||Persistent Mullerian duct syndrome||AR||5||56|
|AMHR2||Persistent Mullerian duct syndrome||AR||6||35|
|AR||Androgen insensitivity, Spinal and bulbar muscular atrophy, X-linked 1, Hypospadias 1, X-linked||XL||138||594|
|ARX||Lissencephaly, Epileptic encephalopathy, Corpus callosum, agenesis of, with abnormal genitalia, Partington syndrome, Proud syndrome, Hydranencephaly with abnormal genitalia, Mental retardation||XL||66||89|
|ATRX||Carpenter-Waziri syndrome, Alpha-thalassemia/mental retardation syndrome, Holmes-Gang syndrome, Juberg-Marsidi syndrome, Smith-Fineman-Myers syndrome, Mental retardation-hypotonic facies syndrome||XL||59||163|
|BCOR||Microphthalmia, syndromic, Oculofaciocardiodental syndrome||XL||38||50|
|CDKN1C||Beckwith-Wiedemann syndrome, IMAGE syndrome||AD||28||81|
|CHD7||Isolated gonadotropin-releasing hormone deficiency, CHARGE syndrome||AD||244||813|
|CYP11A1||Adrenal insufficiency, congenital, with 46,XY sex reversal, partial or complete||AD/AR||14||26|
|CYP11B1*||Adrenal hyperplasia, congenital, due to 11-beta-hydroxylase deficiency, Glucocorticoid-remediable aldosteronism||AD/AR||27||137|
|CYP17A1||Adrenal hyperplasia, congenital, due to 17-alpha-hydroxylase deficiency||AR||35||122|
|CYP19A1||Aromatase deficiency, Aromatase excess syndrome||AR||16||51|
|CYP21A2*||Adrenal hyperplasia, congenital, due to 21-hydroxylase deficiency, Hyperandrogenism, nonclassic , due to 21-hydroxylase deficiency||AR||48||298|
|DHH||46,XY partial gonadal dysgenesis, with minifascicular neuropathy, 46,XY sex reversal 7||AD/AR||4||17|
|DYNC2H1||Short -rib thoracic dysplasia with or without polydactyly type 1, Short -rib thoracic dysplasia with or without polydactyly type 3, Asphyxiating thoracic dysplasia (ATD; Jeune), SRPS type 2 (Majewski)||AR/Digenic||144||109|
|ERCC3||Xeroderma pigmentosum, Trichothiodystrophy, photosensitive||AR||10||18|
|FGFR1||Pfeiffer syndrome, Trigonocephaly, Hypogonadotropic hypogonadism, Osteoglophonic Dwarfism - Craniostenosis, Hartsfield syndrome||AD/Digenic/Multigenic||69||241|
|FIG4||Amyotrophic lateral sclerosis, Polymicrogyria, bilateral occipital, Yunis-Varon syndrome, Charcot-Marie-Tooth disease||AD/AR||29||60|
|GATA4||Tetralogy of Fallot, Atrioventricular septal defect, Testicular anomalies with or without congenital heart disease, Ventricular septal defect, Atrial septal defect||AD||36||126|
|HSD3B2||3-beta-hydroxysteroid dehydrogenase, II deficiency||AR||11||63|
|HSD17B3||17-Beta hydroxysteroid dehydrogenase III deficiency||AR||24||63|
|IRF6||Orofacial cleft, Popliteal pterygium syndrome, van der Woude syndrome||AD||43||337|
|KISS1R||Precocious puberty, central 1||AR||7||35|
|LHCGR||Precocious puberty, male, Leydig cell hypoplasia, Luteinizing hormone resistance, female||AR||33||69|
|MAMLD1||Hypospadias 2, X-linked||XL||5||19|
|MAP3K1||46,XY sex reversal 6||AD||9||22|
|MKRN3||Central precocious puberty||AD||5||30|
|MKS1||Bardet-Biedl syndrome, Meckel syndrome||AR||43||52|
|NR5A1||Adrenocortical insufficiency, Premature ovarian failure, 46,XY sex reversal||AD/AR||26||175|
|NR0B1||Adrenal hypoplasia, congenital, 46,XY sex reversal||XL||73||251|
|POR||Disordered steroidogenesis due to cytochrome p450 oxidoreductase deficiency, Antley-Bixler syndrome||AR||12||68|
|PROK2||Hypogonadism, hypogonadotropic, Kallmann syndrome||AD/AR||7||20|
|RSPO1||Palmoplantar hyperkeratosis with squamous cell carcinoma of skin and 46,XX sex reversal||AR||3||3|
|SOX9||Campomelic dysplasia, 46,XY sex reversal, Brachydactyly with anonychia (Cooks syndrome)||AD||44||141|
|SRD5A2||Steroid 5-alpha-reductase 2 deficiency||AR||38||120|
|SRY||46,XX disorder of sex development, 46,XY disorder of sex development||YL||26||107|
|STAR||Lipoid adrenal hyperplasia||AR||18||81|
|WT1||Denys-Drash syndrome, Frasier syndrome, Wilms tumor, Nephrotic syndrome, type 4||AD||36||175|
|ZFPM2||46,XY sex reversal, Diaphragmatic hernia 3, Tetralogy of Fallot||AD||9||44|
* Some, or all, of the gene is duplicated in the genome. Read more.
# The gene has suboptimal coverage (means <90% of the gene’s target nucleotides are covered at >20x with mapping quality score (MQ>20) reads).
The sensitivity to detect variants may be limited in genes marked with an asterisk (*) or number sign (#)
Gene refers to the HGNC approved gene symbol; Inheritance refers to inheritance patterns such as autosomal dominant (AD), autosomal recessive (AR), X-linked (XL), X-linked dominant (XLD) and X-linked recessive (XLR); ClinVar refers to the number of variants in the gene classified as pathogenic or likely pathogenic in this database (ClinVar); HGMD refers to the number of variants with possible disease association in the gene listed in Human Gene Mutation Database (HGMD). The list of associated, gene specific phenotypes are generated from CGD or Orphanet databases.
Non-coding variants covered by the panel
|Gene||Genomic location HG19||HGVS||RefSeq||RS-number|
Added and removed genes from the panel
|Genes added||Genes removed|
|CYP11A1 DHH ERCC3 FGF8 FGFR1 KISS1R MAMLD1 MAP3K1 MKRN3 PROK2|
Test strengthThe strengths of this test include:
- CAP and ISO-15189 accreditations covering all operations at Blueprint Genetics including all Whole Exome Sequencing, NGS panels and confirmatory testing
- CLIA-certified personnel performing clinical testing in a CLIA-certified laboratory
- Powerful sequencing technologies, advanced target enrichment methods and precision bioinformatics pipelines ensure superior analytical performance
- Careful construction of clinically effective and scientifically justified gene panels
- Our Nucleus online portal providing transparent and easy access to quality and performance data at the patient level
- Our publically available analytic validation demonstrating complete details of test performance
- ~1,500 non-coding disease causing variants in Blueprint WES assay (please see below ‘Non-coding disease causing variants covered by this panel’)
- Our rigorous variant classification based on modified ACMG variant classification scheme
- Our systematic clinical interpretation workflow using proprietary software enabling accurate and traceable processing of NGS data
- Our comprehensive clinical statements
Although the CYP21A2 and CYP11B1 genes have segmentally duplicated pseudogenes that reduce sensitivity of NGS diagnostics in general, Blueprint Genetics custom assay has good coverage (>20x) with high mapping rates (mapping quality >40) for target regions of these genes: CYP21A2 98.6% and CYP11B1 100.0%. Our validation showed mean coverage of 105x for CYP21A2 gene and 244x for CYP11B1. Thus, our NGS Panel is not expected to have major limitations in detecting sequence variants in these genes. Genes with partial, or whole gene, segmental duplications in the human genome are marked with an asterisk (*) if they overlap with the UCSC pseudogene regions. The technology may have limited sensitivity to detect variants in genes marked with these symbols (please see the Panel content table above).This test does not detect the following:
- Complex inversions
- Gene conversions
- Balanced translocations
- Mitochondrial DNA variants
- Repeat expansion disorders unless specifically mentioned
- Non-coding variants deeper than ±20 base pairs from exon-intron boundary unless otherwise indicated (please see above Panel Content / non-coding variants covered by the panel).
This test may not reliably detect the following:
- Low level mosaicism
- Stretches of mononucleotide repeats
- Indels larger than 50bp
- Single exon deletions or duplications
- Variants within pseudogene regions/duplicated segments
The sensitivity of this test may be reduced if DNA is extracted by a laboratory other than Blueprint Genetics.
For additional information, please refer to the Test performance section and see our Analytic Validation.
The Blueprint Genetics abnormal genitalia/ disorders of sex development panel covers classical genes associated with indeterminate sex and pseudohermaphroditism, male pseudohermaphroditism, female pseudohermaphroditism, Androgen insensitivity syndrome, Persistent Mullerian duct syndrome and congenital adrenal hyperplasia. The genes on the panel have been carefully selected based on scientific literature, mutation databases and our experience.
Our panels are sliced from our high-quality whole exome sequencing data. Please see our sequencing and detection performance table for different types of alterations at the whole exome level (Table).
Assays have been validated for different starting materials including EDTA-blood, isolated DNA (no FFPE), saliva and dry blood spots (filter card) and all provide high-quality results. The diagnostic yield varies substantially depending on the assay used, referring healthcare professional, hospital and country. Blueprint Genetics' Plus Analysis (Seq+Del/Dup) maximizes the chance to find a molecular genetic diagnosis for your patient although Sequence Analysis or Del/Dup Analysis may be a cost-effective first line test if your patient's phenotype is suggestive of a specific mutation type.
Performance of Blueprint Genetics Whole Exome Sequencing (WES) assay. All individual panels are sliced from WES data.
|Sensitivity % (TP/(TP+FN)||Specificity %|
|Single nucleotide variants||99.65% (412,456/413,893)||>99.99%|
|Insertions, deletions and indels by sequence analysis|
|1-10 bps||96.94% (17,070/17,608)||>99.99%|
|11-50 bps||99.07% (957/966)||>99.99%|
|Copy number variants (exon level dels/dups)|
|Clinical samples (small CNVs, n=52)|
|1 exon level deletion||92.3% (24/26)||NA|
|2 exons level deletion/duplication||100.0% (11/11)||NA|
|3-7 exons level deletion/duplication||93.3% (14/15)||NA|
|Microdeletion/-duplication sdrs (large CNVs, n=37))|
|Size range (0.1-47 Mb)||100% (37/37)|
|Simulated CNV detection|
|2 exons level deletion/duplication||90.98% (7,357/8,086)||99.96%|
|5 exons level deletion/duplication||98.63% (7,975/8,086)||99.98%|
|The performance presented above reached by WES with the following coverage metrics|
|Mean sequencing depth at exome level||174x|
|Nucleotides with >20x sequencing coverage (%)||99.4%|
The target region for each gene includes coding exons and ±20 base pairs from the exon-intron boundary. In addition, the panel includes non-coding variants if listed above (Non-coding variants covered by the panel). Some regions of the gene(s) may be removed from the panel if specifically mentioned in the ‘Test limitations” section above. The sequencing data generated in our laboratory is analyzed with our proprietary data analysis and annotation pipeline, integrating state-of-the art algorithms and industry-standard software solutions. Incorporation of rigorous quality control steps throughout the workflow of the pipeline ensures the consistency, validity and accuracy of results. Our pipeline is streamlined to maximize sensitivity without sacrificing specificity. We have incorporated a number of reference population databases and mutation databases such as, but not limited, to 1000 Genomes Project, gnomAD, ClinVar and HGMD into our clinical interpretation software to make the process effective and efficient. For missense variants, in silico variant prediction tools such as SIFT, PolyPhen, MutationTaster are used to assist with variant classification. Through our online ordering and statement reporting system, Nucleus, the customer has an access to details of the analysis, including patient specific sequencing metrics, a gene level coverage plot and a list of regions with inadequate coverage if present. This reflects our mission to build fully transparent diagnostics where customers have easy access to crucial details of the analysis process.
We provide customers with the most comprehensive clinical report available on the market. Clinical interpretation requires a fundamental understanding of clinical genetics and genetic principles. At Blueprint Genetics, our PhD molecular geneticists, medical geneticists and clinical consultants prepare the clinical statement together by evaluating the identified variants in the context of the phenotypic information provided in the requisition form. Our goal is to provide clinically meaningful statements that are understandable for all medical professionals regardless of whether they have formal training in genetics.
Variant classification is the corner stone of clinical interpretation and resulting patient management decisions. Our classifications follow the Blueprint Genetics Variant Classification Schemes based on the ACMG guideline 2015. Minor modifications were made to increase reproducibility of the variant classification and improve the clinical validity of the report. Our experience with tens of thousands of clinical cases analyzed at our laboratory allowed us to further develop the industry standard.
The final step in the analysis of sequence variants is confirmation of variants classified as pathogenic or likely pathogenic using bi-directional Sanger sequencing. Variant(s) fulfilling all of the following criteria are not Sanger confirmed: 1) the variant quality score is above the internal threshold for a true positive call, 2) an unambiguous IGV in-line with the variant call and 3) previous Sanger confirmation of the same variant at least three times at Blueprint Genetics. Reported variants of uncertain significance are confirmed with bi-directional Sanger sequencing only if the quality score is below our internally defined quality score for true positive call. Reported copy number variations with a size <10 exons are confirmed by orthogonal methods such as qPCR if the specific CNV has been seen less than three times at Blueprint Genetics.
Our clinical statement includes tables for sequencing and copy number variants that include basic variant information (genomic coordinates, HGVS nomenclature, zygosity, allele frequencies, in silico predictions, OMIM phenotypes and classification of the variant). In addition, the statement includes detailed descriptions of the variant, gene and phenotype(s) including the role of the specific gene in human disease, the mutation profile, information about the gene’s variation in population cohorts and detailed information about related phenotypes. We also provide links to the references used, congress abstracts and mutation databases to help our customers further evaluate the reported findings if desired. The conclusion summarizes all of the existing information and provides our rationale for the classification of the variant.
Identification of pathogenic or likely pathogenic variants in dominant disorders or their combinations in different alleles in recessive disorders are considered molecular confirmation of the clinical diagnosis. In these cases, family member testing can be used for risk stratification within the family. In the case of variants of uncertain significance (VUS), we do not recommend family member risk stratification based on the VUS result. Furthermore, in the case of VUS, we do not recommend the use of genetic information in patient management or genetic counseling. For eligible cases, Blueprint Genetics offers a no charge service to investigate the role of reported VUS (VUS Clarification Service).
Our interpretation team analyzes millions of variants from thousands of individuals with rare diseases. Thus, our database, and our understanding of variants and related phenotypes, is growing by leaps and bounds. Our laboratory is therefore well positioned to re-classify previously reported variants as new information becomes available. If a variant previously reported by Blueprint Genetics is re-classified, our laboratory will issue a follow-up statement to the original ordering health care provider at no additional cost.