Luminex液相芯片技术检测血清MBL在肾移植患者中的应用价值

The value of the Luminex magnetic bead immunoassay in detecting serum MBL in kidney transplantation patients

目的评价Luminex液相芯片磁珠免疫法检测血清甘露糖结合凝集素(MBL)的性能指标,进一步阐明血清MBL检测在肾移植患者中的应用价值。方法回顾性分析2012年2月至2016年5月北京大学第一医院行肾移植手术的患者110例,同时选择同期体检的表观健康人群50名作为正常对照。与传统的ELISA方法比较,评价Luminex磁珠免疫法进行血清MBL检测的精密度、线性范围,同时比较两种方法的相关性。基于Luminex磁珠免疫法检测110例肾移植患者移植前血清MBL,评估不同水平MBL分组肾移植患者移植后发生感染和临床排异事件的频率,采用SPSS 19.0和MedCalc12.7.0进行统计学分析。结果Luminex液相芯片磁珠免疫测定法检测血清MBL重复性精密度≤7.15%,日间精密度≤8.44%,线性范围为0.05~11233.00μg/L;ELISA方法检测血清MBL的线性范围为3.20~4202.70μg/L。Luminex液相芯片磁珠免疫法检测MBL线性范围较宽。相关性分析显示两种方法检测血清MBL具有较好的相关性,回归方程为Y=1.2486X+231.81,相关系数r=0.978(P<0.01);采用Bland-Altman差异分析法显示Luminex方法的测定值高于ELISA方法,平均偏差百分比为37.4%(95%CI 33.7%~41.1%)。检测110例肾移植患者移植前血清MBL中位数(四分位数)为4164.0(2124.0,7064.5)μg/L,将肾移植患者以血清MBL<4164.0μg/L为中/低水平组,以血清MBL≥4164.0μg/L为高水平分组进行比较,血清MBL中/低水平组移植后感染的发生率为47.4%(27/57),高水平组移植后感染的发生率为28.3%(15/53),两组间差异具有统计学意义,χ^2=4.230,P<0.05;血清MBL中/低水平组移植后排异事件的发生率为43.9%(25/57),高水平组移植后排异事件的发生率为20.8%(11/53),两组间差异具有统计学意义,χ^2=6.659,P<0.05。结论Luminex液相芯片磁珠免疫法检测血清MBL线性范围宽,方法学性能优于ELISA方法。肾移植患者在移植前检测血清MBL水平,对于移植后感染和排异事件的预测具有重要价值。

Objective To evaluate the performance of serum mannose-binding lectin(MBL)using Luminex magnetic bead immunoassay,and further clarify the value of serum MBL in the patients of renal transplantation.Methods A retrospective study based on 110 patients who had underwent renal transplantation in Peking University First Hospital from February 2012 to May 2016 was carried out,and 50 healthy persons were selected as controls.The precision,linearity and correlation of serum MBL were evaluated using Luminex magnetic bead immunoassay,and compared with the traditional ELISA method.The frequency of infection and clinical rejection after transplantation was evaluated according to serum pre-transplant MBL level before transplantation,based on the Luminex method.Statistics analysis was implemented with SPSS 19.0 and MedCalc 12.7.0 software.Results The repeatability precision and inter-day precision were less than 7.15%and 8.44%respectively,and linear range was 0.05-11233.00μg/L detected by Luminex immunoassay.The linear range of MBL detected by ELISA was 3.20-4202.70μg/L.The Luminex method had a wider range compared with ELISA.Correlation analysis showed that the regression equation was Y=1.2486X+231.81,and the correlation coefficient was r=0.978(P<0.01).Bland-Altman analysis showed that the average deviation percentage was 37.4%(95%CI 33.7%-41.1%).The median(quartile)of pre-transplant serum MBL was 4164.00(2124.00,7064.50)μg/L.Patients with a serum MBL<4164.00μg/L and MBL≥4164.00μg/L were defined as low-and high-level group,respectively.The incidence of infection among the low-level group and high-level group was 47.4%(27/57)and 28.3%(15/53)respectively,which showed a statistical difference(χ^2=4.230,P<0.05).The incidence of rejection among the low-level group and high-level group was 43.9%(25/57)and 20.8%(11/53)respectively,which also showed a statistical difference(χ^2=6.659,P<0.05).Conclusions The Luminex magnetic bead immunoassay has a wider linearity compared with ELISA in detecting serum MBL.Additionally,serum pre-transplant MBL level has a good predictive value for the infection and rejection reaction after transplantation.