尿样在SELDI技术中的优化研究  

Optimization of SELDI in researching urine sample

作　　者：周怡[1] 廖明[1] 臧宁[1] 罗蓉[1] 何敏[1] 

机构地区：[1]广西医科大学医学科学实验中心,广西南宁530021

出　　处：《现代生物医学进展》2011年第22期4333-4335,共3页Progress in Modern Biomedicine

基　　金：广西科学研究与技术开发计划项目(桂科能05112001-4A;桂科能0630006-5E;桂科能0842009)

摘　　要：目的:建立表面增强激光解吸离子化飞行时间质谱(SELDI-TOF/MS)技术检测尿液样本的方法。方法:采用SELDI技术及弱阳离子交换表面蛋白芯片(CM10)对糖尿病病例组和正常对照组的尿液样本进行分析,从尿液标本的采集、样品保存、上样浓度的控制、实验仪器的内外校准、实验结果重复性验证与分析等方面进行实验条件的优化。结果:反复冻融3次以上的样品经SELDI检测的出峰数量和峰强度情况较差;以1:1或1:2的浓度作倍比稀释后的样本经芯片检测得到的蛋白峰强度和蛋白数量最优;对两组样本重复检测3次,蛋白丰度的平均变异系数(CV值)分别为0.121和0.095;两组样本共检测到202个蛋白峰,其中差异表达蛋白29个,在肝纤维化组中表达上调13个,表达下调16个。结论:初步建立了SELDI技术检测尿液样本的方法,提高了SELDI技术检测尿液样本的质量。Objective： To optimize the method for detecting the urine sample by surface enhanced laser desorption/ionization time of flight mass spectrometry （SELDI-TOF-MS） technique. Methods： SELDI-TOF-MS （PBS II ） and CM10 Protein Chip were used to de- tect the urine samples, which including 4 cases in each of the diabetes group and control group. The collection and preparation of sam- ples, the concentration of the loading samples,the calibration and attendance of laboratory apparatus were optimized. Results： Samples with repeated frost thawing three times or more had the perishing results after SELDI detection. Urine samples with the diluted concentra- tion of 1：1 or 1：2 was best. The corresponding CV values of the experimental and controlling group were 0.121 and 0.095 respectively, which suggested the optimized progect possessed favourable reproducibility. Meanwhile, there were 202 protein peaks detected by SEL- DI-TOF-MS, in 14.4% of which had differential expression including 13 up-regulated and 16 down-regulated protein peaks. Conclusion： The optimized method for the tissue applied in SELDI-TOF-MS was initially established and the experimental quality was also improved.

关 键 词：SELDI-TOF-MS CM10蛋白芯片 肝纤维化组织 

分 类 号：R446[医药卫生—诊断学]