不同方法检测急性移植物抗宿主病患者的血清标志物

Different methods for detecting serum markers in patients with acute graft versus host disease

目的采用两种方法检测急性移植物抗宿主病(acute graft versus host disease,aGVHD)患者的血清生物标志物,筛选出有临床价值的生物标志物,并阐明两种方法检查结果的一致性,建立aGVHD的生物标志物筛查流程。方法收集2016年4月至2018年4月本血液病医学中心进行异基因造血干细胞移植的患者60例,采用蛋白质芯片法和ELISA检测同时期接受异基因造血干细胞移植后发生(aGVHD组,n=30)和未发生急性移植物抗宿主病患者(对照组,n=30)的血清细胞因子表达水平。结果两种不同实验方法检测均发现,aGVHD组患者血清中IL-7、IL-27表达水平高于对照组(P<0.05),而CD127在aGVHD组患者中表达水平降低(P<0.05)。其中IL-27表达水平与急性移植物抗宿主病的疾病严重程度相关(P<0.05)。两种检测方法的ROC曲线下面积均在0.7~0.9之间,差异无统计学意义。结论蛋白质芯片和ELISA两种检测方法的准确度高,结果一致。可以建立先通过蛋白质芯片法筛选出有临床意义的重点细胞因子,再扩大样本量后通过ELISA法进行重复验证的血清生物标志物筛查流程,利于临床开展。

ObjectiveTo clarify the consistency of the results of the 2 methods for detecting serum biomarkers in patients with acute graft versus host disease(aGVHD)in order to screen out biomarkers with clinical value and to establish a biomarker screening process for aGVHD.MethodsSixty patients undergoing allogeneic hematopoietic stem cell transplantation(allo-HSCT)in our medical center from April 2016 to April 2018 were recruited in this study.Protein chip and ELISA were used to detect the serum cytokine levels in the patients with aGVHD(n=30)and those without(control group,n=30).ResultsBoth detection methods showed that the serum levels of IL-7 and IL-27p were significantly higher(P<0.05),while the expression level of CD127 was decreased(P<0.05)in the aGVHD group than the control.The expression level of IL-27 was correlated with the severity of aGVHD(P<0.05).The area under the receiver operating characteristic(ROC)curve was between 0.7 and 0.9,with no significant differences between the 2 methods.ConclusionBoth protein chip and ELISA have high accuracy and consistency in the detection of serum biomarkers in aGVHD patients.It is possible to establish a high-throughput screening of clinically significant cytokines by protein chip,and then expanding the sample size and repeating the validation by ELISA,which is conducive to clinical development.