Tags

Type your tag names separated by a space and hit enter

High-Throughput Screening for CYP21A1P-TNXA/TNXB Chimeric Genes Responsible for Ehlers Danlos Syndrome in Patients with Congenital Adrenal Hyperplasia.

Abstract

Many patients with congenital adrenal hyperplasia (CAH) due to 21 hydroxylase deficiency have CAH-X syndrome, a connective tissue dysplasia consistent with hypermobility type Ehlers Danlos syndrome due to a contiguous gene deletion involving the adjacent CYP21A2 and TNXB genes. CAH-X syndrome is caused by carrying CYP21A1P-TNXA/TNXB chimeric genes (CAH-X CH-1 and CH-2) on one or more alleles. Genetic analysis is cumbersome due to pseudogene interference. We developed a PCR-based CAH-X high-throughput screening method to assess the copy numbers of TNXB exons 35 and 40 that is amenable to either quantitative PCR or droplet digital PCR. The assay was validated in a cohort of 278 subjects from 146 unrelated CAH families. Results were confirmed by a validated Sanger sequencing platform. A total of 44 CAH-X positive calls were made, with 42 (26 CH-1 and 16 CH-2) confirmed. The assay had 100% sensitivity (42 true/42 positives), 99.2% specificity (234 true/236 negatives), and an overall 99.3% accuracy (276/278). Calls made by qPCR and ddPCR were consistent (100%), and ddPCR offered easier data interpretation. The CAH-X prevalence was 15.6% (21/135 probands), higher than the previously estimated 8.5% and was particularly high (29.2% or 21/72) in those with a 30-kb deletion. This assay is suitable for high throughput CAH-X screening, especially for subjects testing positive for CAH in neonatal screening.

Links

  • Publisher Full Text
  • Authors+Show Affiliations

    ,

    National Institutes of Health Clinical Center, Bethesda.

    ,

    National Institutes of Health Clinical Center, Bethesda.

    National Institutes of Health Clinical Center, Bethesda; Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland. Electronic address: dmerke@nih.gov.

    Source

    Pub Type(s)

    Journal Article

    Language

    eng

    PubMed ID

    31229653

    Citation

    Lao, Qizong, et al. "High-Throughput Screening for CYP21A1P-TNXA/TNXB Chimeric Genes Responsible for Ehlers Danlos Syndrome in Patients With Congenital Adrenal Hyperplasia." The Journal of Molecular Diagnostics : JMD, 2019.
    Lao Q, Brookner B, Merke DP. High-Throughput Screening for CYP21A1P-TNXA/TNXB Chimeric Genes Responsible for Ehlers Danlos Syndrome in Patients with Congenital Adrenal Hyperplasia. J Mol Diagn. 2019.
    Lao, Q., Brookner, B., & Merke, D. P. (2019). High-Throughput Screening for CYP21A1P-TNXA/TNXB Chimeric Genes Responsible for Ehlers Danlos Syndrome in Patients with Congenital Adrenal Hyperplasia. The Journal of Molecular Diagnostics : JMD, doi:10.1016/j.jmoldx.2019.06.001.
    Lao Q, Brookner B, Merke DP. High-Throughput Screening for CYP21A1P-TNXA/TNXB Chimeric Genes Responsible for Ehlers Danlos Syndrome in Patients With Congenital Adrenal Hyperplasia. J Mol Diagn. 2019 Jun 20; PubMed PMID: 31229653.
    * Article titles in AMA citation format should be in sentence-case
    TY - JOUR T1 - High-Throughput Screening for CYP21A1P-TNXA/TNXB Chimeric Genes Responsible for Ehlers Danlos Syndrome in Patients with Congenital Adrenal Hyperplasia. AU - Lao,Qizong, AU - Brookner,Brittany, AU - Merke,Deborah P, Y1 - 2019/06/20/ PY - 2019/01/24/received PY - 2019/03/29/revised PY - 2019/06/06/accepted PY - 2019/6/24/entrez PY - 2019/6/24/pubmed PY - 2019/6/24/medline JF - The Journal of molecular diagnostics : JMD JO - J Mol Diagn N2 - Many patients with congenital adrenal hyperplasia (CAH) due to 21 hydroxylase deficiency have CAH-X syndrome, a connective tissue dysplasia consistent with hypermobility type Ehlers Danlos syndrome due to a contiguous gene deletion involving the adjacent CYP21A2 and TNXB genes. CAH-X syndrome is caused by carrying CYP21A1P-TNXA/TNXB chimeric genes (CAH-X CH-1 and CH-2) on one or more alleles. Genetic analysis is cumbersome due to pseudogene interference. We developed a PCR-based CAH-X high-throughput screening method to assess the copy numbers of TNXB exons 35 and 40 that is amenable to either quantitative PCR or droplet digital PCR. The assay was validated in a cohort of 278 subjects from 146 unrelated CAH families. Results were confirmed by a validated Sanger sequencing platform. A total of 44 CAH-X positive calls were made, with 42 (26 CH-1 and 16 CH-2) confirmed. The assay had 100% sensitivity (42 true/42 positives), 99.2% specificity (234 true/236 negatives), and an overall 99.3% accuracy (276/278). Calls made by qPCR and ddPCR were consistent (100%), and ddPCR offered easier data interpretation. The CAH-X prevalence was 15.6% (21/135 probands), higher than the previously estimated 8.5% and was particularly high (29.2% or 21/72) in those with a 30-kb deletion. This assay is suitable for high throughput CAH-X screening, especially for subjects testing positive for CAH in neonatal screening. SN - 1943-7811 UR - https://www.unboundmedicine.com/medline/citation/31229653/High-Throughput_Screening_for_CYP21A1P-TNXA/TNXB_Chimeric_Genes_Responsible_for_Ehlers_Danlos_Syndrome_in_Patients_with_Congenital_Adrenal_Hyperplasia L2 - https://linkinghub.elsevier.com/retrieve/pii/S1525-1578(19)30041-8 DB - PRIME DP - Unbound Medicine ER -