应用SELDI-TOF-MS筛选神经系统先天畸形胎儿羊水诊断标志物

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目的分析神经系统先天畸形胎儿羊水和神经系统正常胎儿羊水的蛋白质表达差异。方法选择8例神经系统畸形胎儿羊水,其中男性4例,女性4例;孕妇年龄21~32岁,平均年龄24.9岁;孕周19~34周,平均孕周23.1周。30例经超声检查神经系统正常胎儿羊水作为对照组,其中男性15例,女性15例;孕妇年龄22~29岁,平均年龄25.1岁;孕周19~22周,平均孕周21.2周。运用表面增强激光解吸离子化飞行时间质谱(SELDI-TOF-MS)蛋白质芯片技术检测神经系统先天畸形胎儿羊水和神经系统正常胎儿羊水蛋白质表达图谱。胎儿羊水用WCX2(弱阳离子交换)芯片检测,采用PBSⅡC型蛋白质芯片阅读机读取数据,Protein-Chip software 3.1软件采集数据,Biomarker Wizard软件分析两组羊水的蛋白质差异。结果 SELDI-TOF-MS技术检测发现神经系统先天畸形胎儿羊水和神经系统正常胎儿羊水的蛋白质存在差异表达,共有9个蛋白质水平发生变化,其中质/荷比4 967.526、5 258.056、11 717.010的差异蛋白质在神经系统异常组表达下调,2 540.415、3 107.119、3 396.759、4 590.965、5 589.200、6 429.417的差异蛋白质在神经系统异常组表达上调。结论神经系统先天畸形胎儿羊水和神经系统正常胎儿羊水之间的蛋白质谱有差异蛋白质表达,检测到9个代表性的差异蛋白质很可能是神经系统疾病胎儿的羊水特异性生物标志物。 Objective To analyze the differences of protein expression between fetal amniotic fluid and normal fetal amniotic fluid in congenital anomalous nervous system. Methods Eight cases of fetal amniotic fluid with abnormal nervous system were selected. There were 4 males and 4 females. The pregnant women were 21 to 32 years old with a mean age of 24.9 years. The gestational age ranged from 19 to 34 weeks and average gestational age was 23.1 weeks. 30 cases of normal fetal nervous system by amniotic fluid as control group, including 15 males and 15 females; pregnant women aged 22 to 29 years, mean age 25.1 years; gestational weeks 19 to 22 weeks, the average gestational age of 21.2 weeks. Protein expression profiles of fetal amniotic fluid and normal fetal amniotic fluid in congenital anomalous fetus of nervous system were detected by using surface enhanced laser desorption / ionization time of flight mass spectrometry (SELDI-TOF-MS) protein chip technology. Fetal amniotic fluid with WCX2 (weak cation exchange) chip detection, using PBS Ⅱ C protein chip reader to read data, Protein-Chip software 3.1 software to collect data, Biomarker Wizard software to analyze protein differences between the two groups of amniotic fluid. Results SELDI-TOF-MS showed that there were differences in the protein expression of fetal amniotic fluid and normal fetal amniotic fluid in the nervous system with congenital malformations. There were 9 proteins in all groups, with a difference of 4 967.526, 5 258.056 and 11 717.010 Proteins were down-regulated in the abnormal group of the nervous system, and the differential proteins of 2 540.415,3 107.119,3 396.759,4 590.965,5 589.200,6 429.417 were up-regulated in the abnormal group of the nervous system. CONCLUSIONS: There are differential protein expressions in protein profiles between the fetal amniotic fluid of the nervous system and normal fetal amniotic fluid. Nine representative differential proteins are likely to be amniotic fluid specific biomarkers of the fetus with neurological diseases.
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