Genotyping in patients with congenital adrenal hyperplasia by sequencing of newborn bloodspot samples

Karissa Ludwig; Fei Lai; Veronica Wiley; Anja Ravine; Shubha Srinivasan

doi:10.1515/jpem-2023-0044

Article

Genotyping in patients with congenital adrenal hyperplasia by sequencing of newborn bloodspot samples

Karissa Ludwig , Fei Lai , Veronica Wiley , Anja Ravine and Shubha Srinivasan

Published/Copyright: September 21, 2023

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From the journal Journal of Pediatric Endocrinology and Metabolism Volume 36 Issue 10

Abstract

Objectives

Genotype–phenotype correlation in congenital adrenal hyperplasia (CAH) caused by 21-hydroxylase deficiency ranges from 45 to 97 %. We performed massively parallel sequencing of CYP21A2 on stored newborn bloodspot samples to catalogue the genotypes present in our patients with CAH and enable genotype–phenotype comparison.

Methods

Participants ≤15 years old with clinically diagnosed CAH were recruited from The Sydney Children’s Hospitals Network. Phenotype was classified from clinical and biochemical details in the medical record as salt wasting (SW), simple virilising (SV), non-classic (NC) or an intermediate phenotype (SW/SV; SV/NC). Amplicon-based sequencing for CYP21A2 was performed on stored newborn bloodspot samples by the New South Wales Newborn Bloodspot Screening Laboratory on MiSeq™Dx (Illumina, California). Available genetic test results were also obtained from the medical records.

Results

Samples from 67 participants (43 % female, age 0.3–15 years) were sequenced, including 9 sibships. SW phenotype was present in 33/67 participants (49 %), SV in 9 (13 %) and NC in 16 (24 %). Intermediate phenotypes included SW/SV in seven participants (10 %) and SV/NC in two (3 %). Variants were identified in 90/116 alleles (78 %). A complete genotype was available in 47/67 participants (70 %). The most common genotype was homozygous c.293-13A/C>G (I2G) in 7/47 participants (15 %). Genotype correlated with the most commonly reported phenotype in 36/44 cases (82 %). Correlation was higher in SW and NC phenotypes.

Conclusions

This study uses genetic testing of newborn bloodspots to identify and characterise the genotypes present in an ethnically diverse Australian population with CAH. It further strengthens our knowledge of genotype–phenotype correlations in CAH.

Keywords: congenital adrenal hyperplasia; genetic diagnosis; genotype; phenotype; newborn bloodspot

Corresponding author: Dr. Karissa Ludwig, Paediatric Endocrinologist, Department of Endocrinology, Queensland Children’s Hospital, 501 Stanley Street, 4101 South Brisbane, QLD, Australia, Phone: 3068 5264, E-mail: karissa.ludwig@health.qld.gov.au

Study performed at Sydney Children’s Hospitals Network, New South Wales, Australia.

Funding source: Pfizer Australasian Paediatric Endocrine Care (APEC)

Acknowledgments

This study was supported by the Sydney Children’s Hospitals Network and the NSW Newborn Screening Programme.

Research ethics: This study protocol was reviewed and approved by the Sydney Children’s Hospitals Network Human Research Ethics Committee (HREC reference: 2018/ETH00679).
Informed consent: Written informed consent was obtained from the parent/legal guardian for all participants.
Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission. Karissa Ludwig – study design, ethics submission, recruitment, data analysis, manuscript preparation. Fei Lai – study design, sample analysis, manuscript preparation. Veronica Wiley – study design, sample analysis, manuscript preparation. Anja Ravine – advised on data analysis, manuscript preparation. Shubha Srinivasan – study design, ethics submission, manuscript preparation.
Competing interests: Authors state no conflict of interest.
Research funding: Study funded by the Pfizer Australasian Paediatric Endocrine Care (APEC) grant. Pfizer had no role in the study design, conduct, data analysis or manuscript preparation.

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Received: 2023-01-28

Accepted: 2023-09-01

Published Online: 2023-09-21

Published in Print: 2023-10-26

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Articles in the same Issue

https://doi.org/10.1515/jpem-2023-0044

Keywords for this article

congenital adrenal hyperplasia; genetic diagnosis; genotype; phenotype; newborn bloodspot