Maccari Francesca, Galeotti Fabio, Mantovani Veronica, Zampini Lucia, Padella Lucia, Rigon Laura, Concolino Daniela, Fiumara Agata, Pascale Elisa, Pittalà Annarita, Galeazzi Tiziana, Monachesi Chiara, Marchesiello Rita Lucia, Coppa Giovanni, Gabrielli Orazio, Volpi Nicola
Department of Life Sciences, University of Modena and Reggio Emilia, Modena, Italy.
Department of Clinical Sciences, Polytechnic University of the Marche, Ancona, Italy.
Anal Biochem. 2018 Sep 15;557:34-41. doi: 10.1016/j.ab.2018.07.007. Epub 2018 Jul 18.
Dried blood spot (DBS) technology is a cheap and easy method largely applied in newborn screening. Mucopolysaccharidoses (MPS) are characterized by the deficit of enzymes that degrade glycosaminoglycans (GAGs) characterized by progressive worsening of the conditions. For a possible early diagnosis of MPS, we developed a method of uronic acid (UA)-GAGs determination in DBS of 600 healthy newborns and from a small group of MPS subjects matched for age. Spotted blood UA-GAGs of the normal newborns are composed of 67.2% chondroitin sulfate (CS), 28.6% heparan sulfate (HS) and 4.4% hyaluronic acid with a CS/HS ratio of 2.35 and a total GAGs content of 0.43 μg/DBS. A chemical evaluation of CS and HS structure was performed by measuring their disaccharide composition, sulfation and the overall charge density. The DBS of four different MPS types presented an increase of total or single UA-GAGs content and/or modifications of the CS and HS disaccharide composition as well as chemical signature also related to the MPS enzymatic defect. The modifications of the UA-GAGs composition, parameters and structure of healthy newborns determined in DBS would be useful for a possible early diagnosis of various MPS types.
干血斑(DBS)技术是一种广泛应用于新生儿筛查的廉价且简便的方法。黏多糖贮积症(MPS)的特征是降解糖胺聚糖(GAGs)的酶缺乏,病情呈进行性加重。为了实现对MPS的早期诊断,我们开发了一种方法,用于测定600名健康新生儿以及一小群年龄匹配的MPS患者干血斑中的糖醛酸(UA)-GAGs。正常新生儿的干血斑UA-GAGs由67.2%的硫酸软骨素(CS)、28.6%的硫酸乙酰肝素(HS)和4.4%的透明质酸组成,CS/HS比值为2.35,总GAGs含量为0.43μg/干血斑。通过测量CS和HS的二糖组成、硫酸化程度和总电荷密度,对CS和HS的结构进行了化学评估。四种不同类型MPS的干血斑显示总UA-GAGs或单一UA-GAGs含量增加,和/或CS和HS二糖组成发生改变,以及与MPS酶缺陷相关的化学特征。在干血斑中测定的健康新生儿UA-GAGs组成、参数和结构的改变,可能有助于各种MPS类型的早期诊断。