流式细胞术外周血白细胞分类方法的建立和性能评价  

Establishment and performance evaluation of peripheral blood leukocyte classification reference method by flow cytometry

作　　者：宋颖 韩姣姣 赵强 徐翀 SONG Ying;HAN Jiaojiao;ZHAO Qiang;XU Chong(Clinical Hematology and Osology Laboratory,Shanghai Center for Clinical Laboratory,Shanghai 200126,China)

机构地区：[1]上海市临床检验中心临床血液体液学研究室,上海200126

出　　处：《检验医学》2022年第12期1190-1195,共6页Laboratory Medicine

基　　金：上海市卫生健康委员会卫生行业临床研究专项(190523195934706)。

摘　　要：目的建立流式细胞术外周血白细胞分类方法,并验证其分类的可行性。方法采用CD16-Pacific Blue、CD45-V500、SYTO16、CD7-PE、CD123-PerCP-Cy5.5、CD14-PE-Cy7、CD3+CD19-APC、CD11b-APC-Cy7抗体组合,建立8色流式细胞术白细胞分类方法。收集50例健康体检者乙二胺四乙酸二钾抗凝外周血样本,分别采用全光谱流式细胞仪(流式法)、全自动血液分析仪(仪器法)和显微镜镜检(手工法)3种方法进行白细胞分类计数。采用Passing&Bablok回归对3种方法检测结果进行两两比较。以手工法为参考方法,评价3种方法白细胞分类方法的一致性。结果成功建立了8色流式细胞术白细胞分类方法。仪器法和流式法的分类结果的回归方程的线性概率值(P)>0.05,2种方法中性粒细胞(NEUT)、淋巴细胞(LYMPH)、单核细胞(MONO)、嗜酸性粒细胞(EOS)、嗜碱性粒细胞(BASO)项目回归方程的斜率分别为1.01、0.98、0.97、1.00、2.33,截距分别为-1.53、0.90、-0.11、0.01、-0.23。手工法和仪器法白细胞分类结果的回归方程的线性概率值(P)>0.05;手工法和流式法NEUT、LYMPH、MONO、EOS、BASO项目回归方程的斜率分别为0.98、0.94、0.68、0.86、0.50,截距分别为-0.96、2.64、2.74、0.29、0.30;手工法和仪器法NEUT、LYMPH、MONO、EOS、BASO项目回归方程的斜率分别为0.99、0.94、0.76、0.93、0.12,截距分别为-1.13、1.79、2.69、0.25、0.26。以白细胞分类参考方法(手工法)为标准,流式法和仪器法白细胞分类结果均在参考区间内。方法学比较结果显示,流式法与仪器法的相关性优于流式法与参考方法。结论成功建立了8色流式细胞术白细胞分类方法。该方法白细胞分类计数结果与参考方法具有相关性,且均在参考区间内。建立的流式法与仪器法的方法学一致性优于参考方法,可进一步提高临床血常规检测中白细胞分类的准确性。Objective To establish the classification reference method of peripheral blood leukocytes by flow cytometry,and to verify the feasibility of its classification.Methods An 8-colour flow cytometry leukocyte counting protocol was established using a combination of CD16-Pacific Blue,CD45-V500,SYTO16,CD7-PE,CD123-PerCP-Cy5.5,CD14-PE-Cy7,CD3+CD19-APC and CD11b-APC-Cy7 antibodies.Peripheral blood samples were collected from 50 healthy subjects with ethylenediaminetetraacetic acid dipotassium anticoagulation,and leukocyte classification was performed by full-spectrum flow cytometry(flow method),fully automated hematology analyzer(instrument method)and microscopy(manual method).Passing&Bablok regression was used to compare the determination results of the 3 methods for paired comparison.The manual method was used as a reference method to evaluate the consistency of the 3 methods of leukocyte classification methods.Results An 8-colour flow cytometry leukocyte counting protocol was established successfully.The linear probability values(P)of regression equations for the classification results of instrument method and flow method were>0.05.The slopes of the regression equations for neutrophil(NEUT),lymphocyte(LYMPH),monocyte(MONO),eosinophil(EOS)and basophil(BASO)of the 2 methods were 1.01,0.98,0.97,1.00 and 2.33,respectively,and the intercepts were-1.53,0.90,-0.11,0.01 and-0.23,respectively.The linear probability values(P)of regression equations for the classification results of manual method and instrument method were>0.05.The slopes of the regression equations for NEUT,LYMPH,MONO,EOS,and BASO of the manual method and flow method were 0.98,0.94,0.68,0.86 and 0.50,respectively,and the intercepts were-0.96,2.64,2.74,0.29 and 0.30,respectively.The slopes of the regression equations for NEUT,LYMPH,MONO,EOS,and BASO by the manual method and instrument method were 0.99,0.94,0.76,0.93 and 0.12,with the intercepts of-1.13,1.79,2.69,0.25 and 0.26,respectively.The slopes of the regression equations for NEUT,LYMPH,MONO,EOS,and BASO by

关 键 词：白细胞分类 流式细胞术 参考方法 

分 类 号：R446.1[医药卫生—诊断学]