毛细管电泳用于高苯丙氨酸血症基因诊断的临床应用研究

Clinical application study on capillary electrophoresis-based gene diagnosis of hyperphenylalaninemia

目的评估一种基于毛细管电泳的高苯丙氨酸血症的快速基因诊断方法的临床价值。方法前瞻性单中心研究。选取2021年2月至2023年2月于南京市妇幼保健院经液相串联质谱新生儿筛查检测疑似高苯丙氨酸血症的40例患儿。其中男22例,女18例;平均诊断年龄21.93 d。采用毛细管电泳技术对40例疑似高苯丙氨酸血症患儿苯丙氨酸羟化酶(PAH)基因中的85个变异位点进行检测,对未明确诊断患儿的PAH基因进一步采用Sanger测序进行检测。评估毛细管电泳技术的检出率、敏感度和特异度。结果在40例疑似高苯丙氨酸血症患儿中,毛细管电泳技术共检出71个PAH变异,32例患儿明确基因诊断,5例患儿仅发现1个致病变异,3例患儿未检出变异。毛细管电泳技术的检出率为80.00%,敏感度为88.75%,特异度为100%。结论毛细管电泳技术能够快速、高效、准确地检出PAH基因变异,且成本较低,是一种极具临床应用前景的高苯丙氨酸血症基因检测方法。

Objective To evaluate the clinical value of a capillary electrophoresis-based method for gene diagnosis of hyperphenylalaninemia.Methods In this single-center prospective study,40 newborns with suspected hyperphenylalaninemia detected by neonatal liquid chromatography-tandem mass spectrometry screening at Nanjing Maternity and Child Health Care Hospital from February 2021 to February 2023 were included,with 22 males,18 females and a mean age at diagnosis of 21.93 days.Capillary electrophoresis was used to detect 85 variants of the phenylalanine hydroxylase(PAH)gene in 40 newborns with suspected hyperphenylalaninemia.The PAH gene of undiagnosed patients was further analyzed by Sanger sequencing.The detection rate,sensitivity and specificity of capillary electrophoresis were calculated.Results Among these 40 newborns with suspected hyperphenylalaninemia,71 PAH variants were detected by capillary electrophoresis,32 patients were clearly diagnosed,only 1 pathogenic variant was found in 5 patients,and no pathogenic variant was found in the last 3 patients.Therefore,the detection rate,sensitivity and specificity of capillary electrophoresis for analysis of the PAH gene were 80.00%,88.75%and 100%,respectively.Conclusions The capillary electrophoresis-based method can rapidly,efficiently and accurately detect PAH gene variants at lower cost and is a promising gene detection method for hyperphenylalaninemia in clinical practice.