摘要
目的建立流式微球分析技术联合检测胱抑素(Cys)C和中性粒细胞明胶酶相关脂质运载蛋白(NGAL)的方法,并初步研究两者在肾移植术后巨细胞病毒(CMV)感染患者血清中的含量变化。方法实验方法的建立包括检测微球包被的效果和稳定性,优化Cys C和NGAL的检测含量范围,确定生物素化抗体浓度,确定PE-链霉亲和素浓度,绘制标准曲线图确定相关系数。采用该法检测对照组(10名健康人)、CMV阴性组(10例肾移植术后CMV-pp65阴性患者)和CMV阳性组(20例肾移植术后CMV-pp65阳性患者)的血清Cys C和NGAL含量。结果成功建立流式微球分析技术联合检测Cys C和NGAL的方法。对照组、CMV阴性组和CMV阳性组的血清NGAL含量分别为8 759(1 465)pg/ml、10 472(856)pg/ml、12 817(4 533)pg/ml,依次升高且差异有统计学意义(P<0.001)。3组间Cys C比较差异无统计学意义(P>0.05)。结论本实验建立流式微球分析技术联合检测Cys C和NGAL标准曲线的方法相关性和重复性良好,并初步验证NGAL含量在肾移植术后CMV感染中明显升高,提示肾移植术后CMV活动性感染可能对肾小管造成急性损伤。
Objective To establish the method of cytometric bead assay in combined detection of cystatin C（ Cys C） and neutrophil gelatinase-associated lipocalin（ NGAL） and to preliminarily investigate the changes in the serum levels of Cys C and NGAL in cytomegalovirus（ CMV）-infected patients after renal transplantation. Methods The experimental method was established as following. The effect and stability of microsphere capsulation were evaluated. The detection range of Cys C and NGAL concentration was improved. The concentrations of biotinylated antibody and PE-streptavidin were quantitatively measured. The correlation coefficient of the standard curve was calculated. The serum levels of Cys C and NGAL in the control（ n = 10, healthy person）, negative CMV（ n = 10, CMV-pp65 negative patients after renal transplantation） and positive CMV groups（ n = 20,CMV-pp65 positive patients after renal transplantation） were detected using this method. Results The method of cytometric bead assay in combined detection of Cys C and NGAL was established successfully. In the control,negative and positive CMV groups,the serum levels of NGAL were 8 759（ 1 465） pg / ml,10 472（ 856） pg / ml and 12 817（ 4 533） pg / ml with statistical significance（ all P〈0. 001）. No statistical significance was noted in terms of Cys C concentration among three groups（ all P〉0. 05）. Conclusions Combined application of cytometricbead assay detects the standard curve of Cys C and NGAL with high relevance and reproducibility. Preliminary results demonstrates that NGAL concentration is significantly elevated in CMV-infected patients following renal transplantation,hinting that CMV active infection probably leads to acute renal tubular injury.
出处
《器官移植》
CAS
CSCD
2016年第3期191-195,共5页
Organ Transplantation
基金
首都卫生发展科研专项(首发2014-2-5071)
