Saville Jennifer T, Fuller Maria
Genetics and Molecular Pathology, SA Pathology at Women's and Children's Hospital, 72 King William Road, North Adelaide, SA 5006, Australia.
Adelaide Medical School, University of Adelaide, North Terrace, Adelaide, SA 5000, Australia.
Metabolites. 2021 Jul 8;11(7):446. doi: 10.3390/metabo11070446.
Following clinical indications, the laboratory diagnosis of the inherited metabolic myopathy, Pompe disease (PD), typically begins with demonstrating a reduction in acid alpha-glucosidase (GAA), the enzyme required for lysosomal glycogen degradation. Although simple in concept, a major challenge is defining reference intervals, as even carriers can have reduced GAA, and pseudodeficiencies complicate interpretation. Here, we developed a mass spectrometric assay for quantification of a urinary glycogen metabolite (tetrasaccharide) and reported on its utility as a confirmatory test for PD in a diagnostic laboratory. Using two age-related reference intervals, eight returned tetrasaccharide concentrations above the calculated reference interval but did not have PD, highlighting non-specificity. However, retrospective analysis revealed elevated tetrasaccharide in seven infantile-onset (IOPD) cases and sixteen late-onset (LOPD) cases, and normal concentrations in one heterozygote. Prospective tetrasaccharide analysis in nine individuals with reduced GAA confirmed IOPD in one, LOPD in six and identified two heterozygotes. Using this metabolite as a biomarker of therapeutic response was not overly informative; although most patients showed an initial drop following therapy initiation, tetrasaccharide concentrations fluctuated considerably and remained above reference intervals in all patients. While useful as a confirmation of PD, its utility as a biomarker for monitoring treatment warrants further investigation.
根据临床指征,遗传性代谢性肌病庞贝病(PD)的实验室诊断通常始于证明酸性α-葡萄糖苷酶(GAA)减少,GAA是溶酶体糖原降解所需的酶。尽管概念简单,但一个主要挑战是确定参考区间,因为即使是携带者的GAA也可能降低,而且假缺陷会使解释变得复杂。在此,我们开发了一种用于定量尿糖原代谢物(四糖)的质谱分析方法,并报告了其在诊断实验室作为PD确证试验的效用。使用两个与年龄相关的参考区间,有8例的四糖浓度高于计算出的参考区间,但并未患庞贝病,这突出了该方法的非特异性。然而,回顾性分析显示,7例婴儿型庞贝病(IOPD)病例和16例晚发型庞贝病(LOPD)病例的四糖水平升高,1例杂合子的浓度正常。对9例GAA降低的个体进行前瞻性四糖分析,确诊1例IOPD,6例LOPD,并识别出2例杂合子。将这种代谢物用作治疗反应的生物标志物并没有提供过多信息;尽管大多数患者在开始治疗后四糖浓度最初有所下降,但四糖浓度波动很大,且所有患者的四糖浓度仍高于参考区间。虽然它作为PD的确证方法很有用,但其作为监测治疗的生物标志物的效用仍需进一步研究。