Nationwide carrier screening for congenital adrenal hyperplasia: integrated approach of CYP21A2 pathogenic variant genotyping and comprehensive large gene deletion analysis
Abstract Background Congenital Adrenal Hyperplasia (CAH) due to 21-hydroxylase deficiency (21-OHD CAH) is an autosomal recessive disorder resulting from pathogenic variants in the CYP21A2 gene. The disorder exhibits variable clinical severity, with the classical form manifesting as salt-wasting cris...
Saved in:
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2025-01-01
|
| Series: | BMC Medical Genomics |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12920-025-02089-5 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832585291856084992 |
|---|---|
| author | Yossawat Suwanlikit Bhakbhoom Panthan Pawares Chitayanan Sommon Klumsathian Angkana Charoenyingwattana Wasun Chantratita Objoon Trachoo |
| author_facet | Yossawat Suwanlikit Bhakbhoom Panthan Pawares Chitayanan Sommon Klumsathian Angkana Charoenyingwattana Wasun Chantratita Objoon Trachoo |
| author_sort | Yossawat Suwanlikit |
| collection | DOAJ |
| description | Abstract Background Congenital Adrenal Hyperplasia (CAH) due to 21-hydroxylase deficiency (21-OHD CAH) is an autosomal recessive disorder resulting from pathogenic variants in the CYP21A2 gene. The disorder exhibits variable clinical severity, with the classical form manifesting as salt-wasting crisis in neonates, while inducing ambiguous genitalia in females and precocious puberty in males through simple virilization. Identifying at-risk couples during the preconception stage holds significance for optimizing reproductive choices. Methods: This study included 204 unrelated preconception individuals undergoing carrier screening. A robust molecular approach was devised for rapid detection of nine prevalent CYP21A2 pathogenic variants, utilizing Amplification-Refractory Mutation System (ARMS) PCR and mass spectrometry (MS) genotyping. Complementary quantitative real-time PCR (qPCR) and PCR-based Restriction Fragment Length Polymorphism (PCR-based RFLP) assays were employed for comprehensive gene deletion analysis. The concordance of pathogenic variant detection between ARMS-PCR and MS, as well as the consistency observed in molecular insights from qPCR and PCR-based RFLP, fortified the accuracy of our methodologies. Results: Our combined method could detect common pathogenic variants and large gene deletions with high concordance between ARMS-PCR, MS genotyping, qPCR, and PCR-based RFLP assays. Remarkably, two carriers exhibited significant large-scale deletions, while another manifested a carrier state due to minor-scale gene conversion. The estimated carrier frequency in our cohort using these methods was approximately 1 in 65 individuals. Conclusions: The methods used for 21-OHD CAH carrier screening offer a reliable, swift, and cost-effective approach for detecting common pathogenic variants and large deletions. Despite some limitations, such as the inability to detect all rare mutations, the techniques provide a practical solution for carrier screening, with an estimated carrier frequency of 1 in 65 in our study population. These findings support the potential adoption of these methods in national carrier screening programs, offering a practical balance between efficiency and affordability. |
| format | Article |
| id | doaj-art-77a3b7f14a614829a9178e30f5626ca5 |
| institution | Kabale University |
| issn | 1755-8794 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Medical Genomics |
| spelling | doaj-art-77a3b7f14a614829a9178e30f5626ca52025-01-26T12:56:55ZengBMCBMC Medical Genomics1755-87942025-01-0118111110.1186/s12920-025-02089-5Nationwide carrier screening for congenital adrenal hyperplasia: integrated approach of CYP21A2 pathogenic variant genotyping and comprehensive large gene deletion analysisYossawat Suwanlikit0Bhakbhoom Panthan1Pawares Chitayanan2Sommon Klumsathian3Angkana Charoenyingwattana4Wasun Chantratita5Objoon Trachoo6Undergraduate Program in Doctor of Medicine, Faculty of Medicine Ramathibodi Hospital, Mahidol UniversityCenter for Medical Genomics, Faculty of Medicine Ramathibodi Hospital, Mahidol UniversityPanthupark Genetics ClinicCenter for Medical Genomics, Faculty of Medicine Ramathibodi Hospital, Mahidol UniversityCenter for Medical Genomics, Faculty of Medicine Ramathibodi Hospital, Mahidol UniversityCenter for Medical Genomics, Faculty of Medicine Ramathibodi Hospital, Mahidol UniversityCenter for Medical Genomics, Faculty of Medicine Ramathibodi Hospital, Mahidol UniversityAbstract Background Congenital Adrenal Hyperplasia (CAH) due to 21-hydroxylase deficiency (21-OHD CAH) is an autosomal recessive disorder resulting from pathogenic variants in the CYP21A2 gene. The disorder exhibits variable clinical severity, with the classical form manifesting as salt-wasting crisis in neonates, while inducing ambiguous genitalia in females and precocious puberty in males through simple virilization. Identifying at-risk couples during the preconception stage holds significance for optimizing reproductive choices. Methods: This study included 204 unrelated preconception individuals undergoing carrier screening. A robust molecular approach was devised for rapid detection of nine prevalent CYP21A2 pathogenic variants, utilizing Amplification-Refractory Mutation System (ARMS) PCR and mass spectrometry (MS) genotyping. Complementary quantitative real-time PCR (qPCR) and PCR-based Restriction Fragment Length Polymorphism (PCR-based RFLP) assays were employed for comprehensive gene deletion analysis. The concordance of pathogenic variant detection between ARMS-PCR and MS, as well as the consistency observed in molecular insights from qPCR and PCR-based RFLP, fortified the accuracy of our methodologies. Results: Our combined method could detect common pathogenic variants and large gene deletions with high concordance between ARMS-PCR, MS genotyping, qPCR, and PCR-based RFLP assays. Remarkably, two carriers exhibited significant large-scale deletions, while another manifested a carrier state due to minor-scale gene conversion. The estimated carrier frequency in our cohort using these methods was approximately 1 in 65 individuals. Conclusions: The methods used for 21-OHD CAH carrier screening offer a reliable, swift, and cost-effective approach for detecting common pathogenic variants and large deletions. Despite some limitations, such as the inability to detect all rare mutations, the techniques provide a practical solution for carrier screening, with an estimated carrier frequency of 1 in 65 in our study population. These findings support the potential adoption of these methods in national carrier screening programs, offering a practical balance between efficiency and affordability.https://doi.org/10.1186/s12920-025-02089-521 hydroxylase deficiencyCongenital adrenal hyperplasiaGenetic carrier screening |
| spellingShingle | Yossawat Suwanlikit Bhakbhoom Panthan Pawares Chitayanan Sommon Klumsathian Angkana Charoenyingwattana Wasun Chantratita Objoon Trachoo Nationwide carrier screening for congenital adrenal hyperplasia: integrated approach of CYP21A2 pathogenic variant genotyping and comprehensive large gene deletion analysis BMC Medical Genomics 21 hydroxylase deficiency Congenital adrenal hyperplasia Genetic carrier screening |
| title | Nationwide carrier screening for congenital adrenal hyperplasia: integrated approach of CYP21A2 pathogenic variant genotyping and comprehensive large gene deletion analysis |
| title_full | Nationwide carrier screening for congenital adrenal hyperplasia: integrated approach of CYP21A2 pathogenic variant genotyping and comprehensive large gene deletion analysis |
| title_fullStr | Nationwide carrier screening for congenital adrenal hyperplasia: integrated approach of CYP21A2 pathogenic variant genotyping and comprehensive large gene deletion analysis |
| title_full_unstemmed | Nationwide carrier screening for congenital adrenal hyperplasia: integrated approach of CYP21A2 pathogenic variant genotyping and comprehensive large gene deletion analysis |
| title_short | Nationwide carrier screening for congenital adrenal hyperplasia: integrated approach of CYP21A2 pathogenic variant genotyping and comprehensive large gene deletion analysis |
| title_sort | nationwide carrier screening for congenital adrenal hyperplasia integrated approach of cyp21a2 pathogenic variant genotyping and comprehensive large gene deletion analysis |
| topic | 21 hydroxylase deficiency Congenital adrenal hyperplasia Genetic carrier screening |
| url | https://doi.org/10.1186/s12920-025-02089-5 |
| work_keys_str_mv | AT yossawatsuwanlikit nationwidecarrierscreeningforcongenitaladrenalhyperplasiaintegratedapproachofcyp21a2pathogenicvariantgenotypingandcomprehensivelargegenedeletionanalysis AT bhakbhoompanthan nationwidecarrierscreeningforcongenitaladrenalhyperplasiaintegratedapproachofcyp21a2pathogenicvariantgenotypingandcomprehensivelargegenedeletionanalysis AT pawareschitayanan nationwidecarrierscreeningforcongenitaladrenalhyperplasiaintegratedapproachofcyp21a2pathogenicvariantgenotypingandcomprehensivelargegenedeletionanalysis AT sommonklumsathian nationwidecarrierscreeningforcongenitaladrenalhyperplasiaintegratedapproachofcyp21a2pathogenicvariantgenotypingandcomprehensivelargegenedeletionanalysis AT angkanacharoenyingwattana nationwidecarrierscreeningforcongenitaladrenalhyperplasiaintegratedapproachofcyp21a2pathogenicvariantgenotypingandcomprehensivelargegenedeletionanalysis AT wasunchantratita nationwidecarrierscreeningforcongenitaladrenalhyperplasiaintegratedapproachofcyp21a2pathogenicvariantgenotypingandcomprehensivelargegenedeletionanalysis AT objoontrachoo nationwidecarrierscreeningforcongenitaladrenalhyperplasiaintegratedapproachofcyp21a2pathogenicvariantgenotypingandcomprehensivelargegenedeletionanalysis |