表面增强激光解析/离子化飞行时间质谱技术在神经管缺陷胎儿母亲血清标志物筛查中的应用

Application of surface enhanced laser desorption/ionization time-of-flight mass spectrometry in screening of markers in maternal serum with fetuses of neural tube defects

目的：探讨表面增强激光解析／离子化飞行时间质谱（SELDI—TOF—MS）技术在筛选神经管缺陷胎儿母亲血清蛋白质标志物中的应用。方法：经超声确诊为胎儿神经管缺陷的孕妇17例和正常孕妇14例，神经管缺陷包括脊柱裂7例、无脑儿4例、脑膨出6例。所有孕妇均于清晨空腹取外周静脉血5ml，离心后取血清，然后用CM10芯片按照蛋白质芯片标准操作流程进行检测。采用PBSIIC型蛋白质芯片阅读机读取数据，设定优化相对分子质量范围1000～30000Da。统计分析使用Ciphergen protein chip3．1．1软件。结果：所有神经管畸形胎儿母亲血清标本共检测到55个差异蛋白峰，其中12个与正常组比较存在统计学意义，包括8种蛋白质／多肽高表达，质荷比（m／z）分别为4105、4297、4188、6650、8583、3282、2750、3327；4种蛋白质／多肽低表达，m／z分别为5497，28078，9155，9434。建立决策分类树，得到了带有3个终结点的决策分类树，获得了2个标志性蛋白质，m／z分别为4105、7788。神经管缺陷诊断模型的灵敏度为88．2％，特异度为100％。结论：应用SELDI技术可以筛查出神经管缺陷的特异性蛋白质标志物，是神经管缺陷产前诊断的重要方法之一。

Objective： To explore the application of surface enhanced laser desorption/ionization time -of- flight mass spectrometry （SELDI-TOF- MS） in screening of protein markers in maternal serum with fetuses of neural tube defects. Methods： 17 pregnant women whose fetuses were diagnosed as neural tube defects by ultrasound （7 cases of spina bifida, 4 cases of anencephaly fetus and 6 cases of encephalocele） and 14 normal pregnant women were selected, 5 ml peripheral venous blood of all the pregnant women were obtained early in the morning, then serum was purified from blood by centrifugation, and SELDI - TOF - MS was detected by PBS II C protein chip biosys- tern, the optimal detection range of molecular weight was 1 000 -30 000Da. The data were analysed by Ciphergen protein chip 3.1.1 software. Results： 55 different protein peaks in maternal serum samples were detected, there was significant difference in 12 protein peaks between study group and control group, including 8 kinds of high expression proteins/peptides, their m/z were 4 105, 4 297, 4 188, 6 650, 8 583, 3 282, 2 750 and 3 327, respectively; 4 kinds of low expression proteins/peptides, their m/z were 5 497, 28 078, 9 155 and 9 434, respectively. 3 decision trees with terminal node and 2 protein markers were gained, whose m/z were 4 105 and 7 788, respectively. The specificity and sensitivity of diagnostic model of neural tube defects were 100% and 88. 2%, respectively. Conclusion ： SELDI - TOF - MS can screen the specific protein markers of neural tube defects, which is one of the important prenatal diagnostic methods.