摘要
目的建立人类白细胞抗原(HLA)混合试剂筛查实验结果判断的界限值,并以HLA单抗原试剂确认实验结果进行验证。方法回顾性分析苏州大学附属第一医院HLA配型实验室2017年10月至2021年12月2255份采用HLA混合试剂进行HLA抗体检测的血清样本结果。其中1139份同期采用HLA单抗原试剂-Ⅰ类试剂检测,1116份同期采用HLA单抗原试剂-Ⅱ类试剂检测。针对两种试剂包被的相同抗原,录入混合试剂中12个HLA-Ⅰ类微珠及5个HLA-Ⅱ类微珠的平均荧光强度(MFI)、标准化背景(NBG)比值及HLA单抗原试剂检测到的77种抗HLA-Ⅰ类抗体及35种抗HLA-Ⅱ类抗体的MFI值。以混合试剂各微珠包被抗原对应的单抗原试剂检测到抗体的阳性率将1139份及1116份样本中各混合试剂检测到的微珠的MFI值及NBG比值进行范围分段,并采用HLA单抗原试剂确认实验对HLA混合试剂筛查实验结果进行验证。结果建立了HLA混合试剂筛查实验17个微珠MFI值及NBG比值判断界限值表,HLA混合试剂1~17号微珠的MFI值为26.86~21925.58,NBG比值为0~434.65。按照包被抗原对应单抗原试剂的阳性检出率,将混合试剂各微珠的MFI值及NBG比值分为阳性区间、可疑阳性区间、可疑阴性区间、阴性区间。HLA混合试剂和HLA单抗原试剂对HLA-Ⅰ类抗体阳性检出率分别为87.5%(997/1139)、66.3%(755/1139);对HLA-Ⅱ类抗体阳性检出率分别为63.4%(707/1116)、44.9%(501/1116)。HLA混合试剂检测HLA-Ⅰ类抗体结果为可疑阴性、可疑阳性、阳性样本中,HLA单抗原试剂的阳性检出率分别为14.9%(17/114)、41.3%(145/351)、91.3%(590/646);HLA混合试剂检测HLA-Ⅱ类抗体结果为可疑阴性、可疑阳性、阳性样本中,HLA单抗原试剂的阳性检出率分别为15.5%(58/375)、26.5%(81/306)、88.8%(356/401)。结论本研究建立了HLA混合试剂筛查实验判断各抗原微珠MFI值及NBG比值的界限值,并经HLA单抗原试剂确认实验进行验证。HLA混合试
Objective To establish the threshold value of human leukocyte antigen(HLA)mixed antigen reagent screening test results,and to verify it by HLA single antigen reagent confirmation test results.Methods The results of 2255 serum samples tested for HLA antibodies by HLA mixed antigen reagent in the department of HLA Laboratory,the First Affiliated Hospital of Soochow University from October 2017 to December 2021 were retrospectively analyzed.Among them,1139 samples were also tested by single antigen HLA Class-Ⅰreagent and 1116 samples were also tested by single antigen HLA Class-Ⅱreagent.Based on the same antigens coated with both reagents,the Mean Fluorescence Intensity(MFI)and Nomalized Background ratio(NBG ratio)of 12 HLA Class-Ⅰbeads and 5 HLA Class-Ⅱbeads in the HLA mixed antigen reagent and the MFI of 77 anti-HLA class-Ⅰantibodies and 35 anti-HLA class-Ⅱantibodies detected by HLA single antigen reagent were recorded.The MFI and NBG ratio of HLA mixed antigen reagent beads in 1139 or 1116 samples were segmented according to the positive rate of antibodyies detected by the single antigen reagent corresponding to the antigens coated with each HLA mixed antigen reagent bead,and the results of the HLA mixed antigen screening test were verified by the HLA single antigen reagent confirmation test.Results The threshold values of MFI and NBG ratio of HLA mixed antigen reagent′s 17 beads were established.The MFI of No.1 to No.17 beads of HLA mixed antigen reagent ranged from 26.86 to 21925.58,and the NBG ratio ranged from 0 to 434.65.According to the positive detection rate of HLA single antigen reagent corresponding to the coated antigens,the MFI and NBG ratio of the beads of HLA mixed antigen reagent were divided into positive interval,suspicious positive interval,suspicious negative interval and negative interval.The positive rates of anti-HLA class-Ⅰantibodies by HLA mixed antigen reagent and single antigen HLA Class-Ⅰreagent were 87.5%(997/1139)and 66.3%(755/1139).The positive rates of anti-HLA cla
作者
陈璐瑶
鲍晓晶
袁晓妮
郁玲莹
何军
Chen Luyao;Bao Xiaojing;Yuan Xiaoni;Yu Lingying;He Jun(HLA Laboratory of Jiangsu Institute of Hematology,the First Affiliated Hospital of Soochow University,Suzhou 215031,China)
出处
《中华医学杂志》
CAS
CSCD
北大核心
2024年第11期857-864,共8页
National Medical Journal of China
基金
国家自然科学基金(82070180)
江苏高校血液学协同创新中心项目(SX21100121)
关键词
人类白细胞抗原
抗体
界限值
移植
Human leukocyte antigen
Antibodies
Threshold value
Transplantation
