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目的建立用荧光底物测定艾杜糖-2-硫酸酯酶(IDS)活性的方法,应用该方法对黏多糖贮积症Ⅱ型(MPSⅡ)病例进行临床鉴别诊断,对高危妊娠的孕妇进行产前诊断。方法采用人工合成的荧光底物4-甲基伞形酮-α-艾杜糖苷-2-硫酸,测定 MPSⅡ疑似患儿血浆中 IDS 活性,并用绒毛或经培养的羊水细胞为检材,对高危妊娠的孕妇进行产前诊断。同时观察孕妇血浆 IDS 活性变化,用基因诊断方法检测胎儿性别。结果获得中国正常人 IDS 活性正常值:血浆中为240.2～668.2nmol/(4 h·ml),绒毛组织为37.2～54.9 nmol/(4 h·mg 蛋白),经培养的羊水细胞为21.4～74.4nmol/(4 h·mg 蛋白);患者血浆的 IDS 活性为0.3～18.6 nmol/(4 h·ml),肯定杂合子的血浆酶活性为88.7～547.9 nmol/(4 h·ml)。在50例疑似患儿中确诊了46例 MPSⅡ患者。10例产前诊断中检出4例男性胎儿患病,5例为女性胎儿。结论荧光法测定 IDS 活性是敏感、可靠、快速的方法,可用于病例鉴别诊断及孕早期 MPSⅡ高危妊娠的产前诊断。
OBJECTIVE: To establish a method for the determination of idose-2-sulfatase (IDS) activity using fluorescent substrate. The method was applied to diagnose mucopolysaccharidosis type Ⅱ (MPSⅡ) in patients with high-risk pregnancies Pre-diagnosis. Methods The artificial substrate of 4-methylumbelliferone-α-iduronoside-2-sulfate was used to measure the plasma IDS activity of MPSⅡ, and to detect the high-risk Pregnant women for prenatal diagnosis. At the same time observe the changes of plasma IDS activity of pregnant women, fetal genital detection using genetic testing methods. Results The normal IDS activity of Chinese normal people was 240.2 ~ 668.2nmol / (4 h · ml) in plasma and 37.2 ~ 54.9 nmol / (4 h · mg protein) in chorionic villi. The cultured amniotic fluid cells were 21.4 ~ 74.4 nmol / (4 h · mg protein). The plasma IDS activity was 0.3 ~ 18.6 nmol / (4 h · ml), and the plasma enzyme activity of the positive heterozygotes was 88.7-547.9 nmol / (4 h · ml). Forty-six MPS II patients were diagnosed in 50 suspected children. Four prenatal diagnoses were detected in 10 male fetuses and 5 female fetuses. Conclusion Fluorescence spectrophotometry is a sensitive, reliable and rapid method for the determination of IDS activity. It can be used in the differential diagnosis of cases and prenatal diagnosis of MPS Ⅱ high risk pregnancy in early pregnancy.