使用自主研发的基于5'核酸外切酶的检测方法对性发育障碍患者进行SRY和DYS14基因快速检测的临床适用性。

Nadar Ruchi, Phadke Nikhil, Khatod Kavita, Khadilkar Vaman, Khadilkar Anuradha V

J Pediatr Endocrinol Metab. 2014 Sep;27(9-10):869-72. doi: 10.1515/jpem-2013-0416.

BACKGROUND

Life threatening conditions are associated with atypical genitalia in newborns. Analysis of genetic sex provides a clue to the underlying etiology in newborns with disorders of sex development (DSD) and can guide further endocrine investigations. Rapid diagnosis of genetic sex would be immensely useful in this situation. Traditionally used methods such as karyotype and fluorescence in situ hybridisation are time-consuming.

OBJECTIVES

To study the clinical applicability of an indigenously developed rapid real-time polymerase chain reaction (RT-PCR) assay for the sex determining region on the Y chromosome (SRY gene) and the DYS14 locus in newborns with DSD.

METHODS

Clinical examination, endocrinological tests, RT-PCR analysis of SRY and DYS14 and karyotype was performed in 15 newborns with DSD.

RESULTS

RESULTS of PCR were available within 4 h. Based on this report, in SRY/DYS14 positive cases, further tests for assessment of testicular function were done. In SRY negative cases, tests for congenital adrenal hyperplasia were done. On comparing PCR results with other tests, the Y chromosome was present on karyotype and testicular tissue was detected by endocrinological and/or histological methods in all (8/15) SRY positive cases. The SRY and DYS14 negative cases (7/15) did not have Y chromosome in the karyotype. Congenital adrenal hyperplasia (CAH) was the most common diagnosis in this group.

CONCLUSIONS

The indigenously developed PCR for dual Y chromosome markers is rapid and sensitive. Further endocrine evaluation of newborns with DSD can be based on these results. Information of genetic sex partly allays the psychosocial distress associated with the condition.

背景

危及生命的情况与新生儿生殖器异常有关。对遗传性别进行分析可为患有性发育障碍（DSD）的新生儿潜在病因提供线索，并可指导进一步的内分泌检查。在这种情况下，快速诊断遗传性别将非常有用。传统使用的方法如核型分析和荧光原位杂交耗时较长。

目的

研究一种自主研发的快速实时聚合酶链反应（RT-PCR）检测方法在检测患有DSD的新生儿Y染色体性别决定区（SRY基因）和DYS14位点的临床适用性。

方法

对15例患有DSD的新生儿进行临床检查、内分泌测试、SRY和DYS14的RT-PCR分析以及核型分析。

结果

PCR结果在4小时内可得。根据该报告，对于SRY/DYS14阳性病例，进行了进一步评估睾丸功能的测试。对于SRY阴性病例，进行了先天性肾上腺皮质增生症的检测。将PCR结果与其他检测结果进行比较时，所有（8/15）SRY阳性病例的核型中均存在Y染色体，并且通过内分泌和/或组织学方法检测到睾丸组织。SRY和DYS14阴性病例（7/15）的核型中没有Y染色体。先天性肾上腺皮质增生症（CAH）是该组中最常见的诊断。