Johnson Britt, Mascher Hermann, Mascher Daniel, Legnini Elisa, Hung Christina Y, Dajnoki Angela, Chien Yin-Hsiu, Maródi László, Hwu Wuh-Liang, Bodamer Olaf A
Division of Clinical and Translational Genetics, Department of Human Genetics, University of Miami-Miller School of Medicine, 1501 NW 10th Ave, Miami, FL 33136, USA.
Ann Lab Med. 2013 Jul;33(4):274-8. doi: 10.3343/alm.2013.33.4.274. Epub 2013 Jun 24.
Recently, lyso-globotriaosylsphingosine (lyso-Gb3) was found to be elevated in plasma of treatment naive male patients and some female patients with Fabry Disease (FD). This study tested whether lyso-Gb3 could be analyzed in dried blood spots (DBS) from filter cards and whether concentrations are elevated in newborn infants with FD. Lyso-Gb3 concentrations were analyzed in DBS following extraction using a novel HPLC-mass spectrometry (MS)/MS method. Lyso-Gb3 levels in DBS were above the lower limit of quantitation (0.28 ng/mL) in 5/17 newborn FD infants (16 males; range: 1.02-8.81 ng/mL), but in none of the newborn controls, in all 13 patients (4 males) with classic FD (range: 2.06-54.1 ng/mL), in 125/159 Taiwanese individuals with symptomatic or asymptomatic FD who carry the late onset α-galactosidase A (GLA) mutation c.936+919G>A (IVS4+919G>A) (3.75±0.69 ng/mL; range: 0.418-3.97 ng/mL) and in 20/29 healthy controls (0.77±0.24 ng/mL; range: 0.507-1.4 ng/mL). The HPLC-MS/MS method for analysis of lyso-Gb3 is robust and yields reproducible results in DBS in patients with FD. However, concentrations of lyso-Gb3 were below the limit of quantitation in most newborn infants with FD rendering this approach not suitable for newborn screening. In addition, most females with the late onset mutation have undetectable lyso-Gb3 concentrations.
最近发现，在未经治疗的男性法布里病（FD）患者以及部分女性患者的血浆中，溶血型球三糖神经酰胺（lyso-Gb3）水平升高。本研究检测了是否能够对滤纸片干血斑（DBS）中的lyso-Gb3进行分析，以及FD新生儿的lyso-Gb3浓度是否升高。采用一种新型高效液相色谱-质谱联用（MS）/MS方法对提取后的DBS中的lyso-Gb3浓度进行分析。在17例FD新生儿（16例男性；范围：1.02 - 8.81 ng/mL）中，有5例DBS中的lyso-Gb3水平高于定量下限（0.28 ng/mL），而所有新生儿对照中均未检测到；在所有13例典型FD患者（4例男性；范围：2.06 - 54.1 ng/mL）中均检测到；在159例携带晚期发病α-半乳糖苷酶A（GLA）突变c.936 + 919G>A（IVS4 + 919G>A）的有症状或无症状的台湾FD患者中，125例检测到（3.75±0.69 ng/mL；范围：0.418 - 3.97 ng/mL）；在29例健康对照中，20例检测到（0.77±0.24 ng/mL；范围：0.507 - 1.4 ng/mL）。用于分析lyso-Gb3的HPLC-MS/MS方法性能稳定，在FD患者的DBS中可产生可重复的结果。然而，大多数FD新生儿的lyso-Gb3浓度低于定量下限，这使得该方法不适用于新生儿筛查。此外，大多数携带晚期发病突变的女性lyso-Gb3浓度无法检测到。
