应用金芯片技术预测肾功能损害的临床应用研究

Research on the clinical application of the prediction of early kidney injury by Au protein chip technology

目的探讨利用金(Au)芯片检测尿蛋白质指纹图谱在预测肾功能损害中的应用价值。方法利用表面增强激光解吸电离飞行时间质谱(SELDI-TOF-MS)技术及Au芯片检测186例肾病患者和188例对照者尿蛋白质指纹图,分析肾病患者与对照组尿蛋白质表达差异并筛选标志蛋白质,结合人工神经网络(ANN)技术建立智能预测模型,评价其诊断肾病的应用价值。对部分差异蛋白质通过电喷雾四级杆飞行时间质谱(ESI-Q-TOF)进行鉴定。用免疫散射比浊法检测尿微球蛋白并与蛋白质指纹图谱结果比较,评价SELDI-TOF-MS检测性能。结果肾病患者与对照者尿中共检测到214个蛋白质峰,有69个蛋白质峰差异有统计学意义(P<0.05),筛选其中质荷比(m/z)11 735、23 770、51 720、58 720、67 650、80 045、91 240蛋白质建立的ANN模型预测肾功能损害的灵敏度为98.8%(85/86),特异度为96.6%(85/88)。蛋白鉴定结果表明m/z 11 735、23 770、67 650、80 045蛋白质分别为β2-微球蛋白、α1-微球蛋白、白蛋白、转铁蛋白。SELDI-TOF-MS检测微球蛋白较免疫散射比浊法更灵敏。结论基于Au蛋白芯片技术的尿蛋白质指纹图谱检测,可快速、灵敏、高通量的显示尿蛋白质表达情况,对诊断肾损伤、判断蛋白尿类型及治疗评价具有重要的应用价值。

Objective To determine urine protein profilings by Au chip and discuss the clinical significance in sensitivity and the rapid prediction of renal function.Methods The study collected 186 patients with nephropathy and 188 controls.The urine protein profilings were determined by surface-enhanced laser desorption-ionization time of flight mass spectrometry（SELDI-TOF-MS） and Au chip.The expression differences of urine protein and protein markers were analyzed and screened between the patients and controls.An artificial neural network（ANN） pattern was developed to evaluate the application significance.The differential protein markers were primarily identified by electrospectrometry ionization quadrupole time of flight mass spectrometry（ESI-Q-TOF-MS）.The results of urine microglobulin detected by immune scatter turbidimetry and protein profilings detected by SELDI-TOF-MS were comparatively analyzed.Results Total of 214 protein peaks and 69 distinguished peaks（P0.05） were obtained from the urine samples of patients with nephropathy and the controls.The peaks with mass-to-charge ratio（m/z） 11 735,23 770,51 720,58 720,67 650,80 045 and 91 240 were screened by ANN.The sensitivity was 98.8%（85/86） and the specificity was 96.6%（85/88）.4 specific protein peaks（m/z at 11 735,23 770,67 650 and 80 045）were identified and considered as β2-microglobulin,α1-microglobulin,albumin and transferrin,respectively.The SELDI-TOF-MS was more sensitive than immune scatter turbidimetry.Conclusions Au chip is a fast,sensitive and high-through method to detect urine protein profilings from patients with nephropathy.It has the application significance in the diagnosis of nephropathy,the identification of albuminuria and the evaluation of renal injury.