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High-Throughput Screening for CYP21A1P-TNXA/TNXB Chimeric Genes Responsible for Ehlers-Danlos Syndrome in Patients with Congenital Adrenal Hyperplasia.
J Mol Diagn 2019; 21(5):924-931JM

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 chimera 1 (CH-1) and chimera 2 (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; this method is amenable to either real-time quantitative PCR or droplet digital PCR (ddPCR). 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 real-time quantitative PCR 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 in subjects testing positive for CAH in neonatal screening.

Authors+Show Affiliations

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

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, vol. 21, no. 5, 2019, pp. 924-931.
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;21(5):924-931.
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, 21(5), pp. 924-931. 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;21(5):924-931. 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 - 2019/06/21/ PY - 2019/01/24/received PY - 2019/03/29/revised PY - 2019/06/06/accepted PY - 2020/09/01/pmc-release PY - 2019/6/24/pubmed PY - 2019/6/24/medline PY - 2019/6/24/entrez SP - 924 EP - 931 JF - The Journal of molecular diagnostics : JMD JO - J Mol Diagn VL - 21 IS - 5 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 chimera 1 (CH-1) and chimera 2 (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; this method is amenable to either real-time quantitative PCR or droplet digital PCR (ddPCR). 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 real-time quantitative PCR 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 in 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 -