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Licensed Unlicensed Requires Authentication Published by De Gruyter May 24, 2022

Validation of steroid ratios for random urine by mass spectrometry to detect 5α-reductase deficiency in Vietnamese children

Thi Chi Mai Tran, Thi Ngoc Anh Tran, Hoang Bich Nga Le, Viet Hoa Nguyen, Minh Dien Tran, Chi Dung Vu and Ronda F. Greaves

Abstract

Objectives

The 5α-reductase-type-2 deficiency (5ARD2) is a rare autosomal recessive 46,XY disorder of sex development caused by the mutated 5α-reductase type 2 (SRD5A2) gene. In this disease, defective conversion of testosterone to dihydrotestosterone leads to variable presentations of male ambiguous genitalia during fetal development. We aimed to examine characteristics of patients presenting with 5ARD2 over a 4 year period.

Methods

Random urine samples of control and patients with suspected 5ARD2 were collected and urine steroidomic metabolites were measured by Gas chromatography-mass spectrometry (GC-MS) in the period from 2017 to 2021 at National Children’s Hospital, Hanoi Vietnam. 5α- to 5β-reduced steroid metabolite ratio, 5a-tetrahydrocortisol to tetrahydrocortisol (5α-THF/THF), was reviewed by receive operator characteristics (ROC) curve analysis. Molecular testing was offered to 25 patients who were diagnosed with 5ARD2 by GC-MS urinary steroid analysis.

Results

Urine steroidomic profiling was conducted for 104 male controls and 25 patients between the ages of 6 months and 13 years old. Twelve of the twenty-five 5ARD2 patients agreed to undertake genetic analysis, and two mutations of the SRD5A2 gene were detected in each patient, confirming the diagnosis. All patients showed a characteristically low ratio of 5α-THF/THF. There was no overlap of 5α-THF/THF ratio values between control and 5ARD2 groups. The ROC of 5α-THF/THF ratio at 0.19 showed 100% sensitivity and 100% specificity for boys between 6 months and 13 years of age.

Conclusions

Analysis of the urine steroid metabolome by GC-MS can be used to assist in the diagnosis of 5ARD2. We recommend consideration of random urine steroid analysis as a first-line test in the diagnosis of 5ARD2.


Corresponding author: Dr. Tran Thi Chi Mai, Faculty of Medical Technology, Hanoi Medical University, Hanoi Vietnam; and Vietnam National Children’s Hospital, Hanoi, Vietnam, Phone: +84934220994, E-mail:
Tran Thi Chi Mai Tran and Thi Ngoc Anh Tran contributed equally to this work.

Funding source: Asia Pacific Federation of Clinical Chemistry and Laboratory Medicine project grant.

Acknowledgments

We sincerely thank the children and their families who agreed to participate in this study. We acknowledge the Asia-Pacific Federation of Clinical Biochemistry and Laboratory Medicine for funding of the urine steroid metabolome method development. We also would like to thank the TRAC (Training and Research Academic Collaboration) Sweden-Vietnam for funding the SRD5A2 gene mutation analysis study.

  1. Research funding: Asia Pacific Federation of Clinical Chemistry and Laboratory Medicine project grant.

  2. Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.

  3. Competing interests: Authors state no conflict of interest.

  4. Informed consent: Written informed consent was obtained from all individuals included in this study.

  5. Ethical approval: Ethics approval for collection and analysis of urine samples was obtained from the Ethics Committee of the Vietnam National Children’s Hospital, Hanoi Vietnam.

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Supplementary Material

The online version of this article offers supplementary material (https://doi.org/10.1515/cclm-2022-0272).


Received: 2022-03-22
Accepted: 2022-05-11
Published Online: 2022-05-24
Published in Print: 2022-07-26

© 2022 Walter de Gruyter GmbH, Berlin/Boston

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