血细胞分析仪快速检测外周血造血干细胞与祖细胞的临床意义

Clinical value evaluation of rapid detection method for the hematopoietic stem/progenitor cell of peripheral blood using the automated hematology analyzer

目的:探讨血细胞分析仪快速检测外周血造血干细胞/祖细胞(hematopoietic stem/progenitor cell,HPC)的临床应用价值。方法:方法学验证和回顾性分析。收集2015年1月至2018年12月期间来福建医科大学附属协和医院就诊的4例为初治急性髓系白血病患者外周血、1名健康供者外周血干细胞采集物5倍稀释液,用于方法学验证;23例自体造血干细胞移植患者、22例异基因外周血造血干细胞移植健康供者的外周静脉血和外周血干细胞采集物5倍稀释液,用于一致性回顾分析。线性试验取已知CD34+细胞含量的外周血、血细胞分离机收集的外周血造血干细胞/祖细胞液各1份,做接近预期上限的高值样本(H),一份低值样本即生理盐水(L),倍比稀释,检测HPC,结果回归分析。一致性试验标本126份,EDTA-K 2抗凝静脉血78份,外周血干细胞采集物48份。同时采集的静脉血,一管血常规和HPC检测,另一管流式细胞术(FCM)检测CD34+细胞。干细胞采集物用灭菌生理盐水5倍稀释后分两管,一管全血细胞计数和HPC计数,另一管FCM检测CD34+细胞。对两种仪器检测结果作比对分析及偏差评估。评价血细胞分析仪对外周血造血干细胞/祖细胞检测的本底计数、携带污染率、精密度、线性范围和与流式细胞术的一致性。结果:本底计数结果为0×109个/L、携带污染率为0.1%符合血细胞分析仪质量要求,重现性不精密度是4.7%~18.8%,HPC线性范围是0~27.201×109个/L、采集液5倍稀释后线性范围是0~0.878×109个/L。血细胞分析仪与FCM分别检测126份样本的结果作比对分析,相关系数r2=0.9601。当WBC>10×109个/L时,两种仪器检测结果具有良好的一致性,斜率位于0.95~1.05之间,医学决定水平处估计的预期偏差<30%。结论:血细胞分析仪检测HPC与FCM比对,具有良好的相关性;当WBC>10×109个/L时,与FCM不仅具有较好的一致性,而且预期偏差符合临床要求;其检测HPC标本无需预处理。

Objective To explore the performance and clinical application value of a rapid detection method for the hematopoietic stem/progenitor cell of peripheral blood using the automated hematology analyzer.Methods Methodology validation and retrospective study.Collected sample from Fujian Medical University Union Hospital from January 2015 to December 2018,the peripheral blood of 4 patients with acute myeloid leukemia was first treated,and one healthy donor′s peripheral blood stem cell collection 5 times diluent,for the methodology validation.And the peripheral venous blood and 5-fold dilutions of peripheral blood stem cell collection,from 23 patients with hematopoietic stem cell transplantation and 22 healthy donors of allogeneic peripheral blood stem cell transplantation,used for consistency retrospective analysis.In the linear test,each of the peripheral blood and HPC collecting solutions from blood cell separator,which known CD34+cell concentration,that was high-value samples for the expected upper limit(H).Another low-value sample is normal saline(L).According to the multiple proportion dilution,HPC was detected and regressed consistency test specimens were 126,EDTA-K2 anticoagulant venous blood 78 and peripheral blood stem cell 48.Venous blood was collected at the same time,one tube of blood routine and HPC detection,the other tube flow cytometry(FCM)detection of CD34+cells.Stem cell collection was diluted 5 times with sterilized saline and divided into two tubes.One tube was used to count whole blood cells and HPC,the other tube was used to detect CD34+cells by FCM.The test results of the two instruments were compared,and the deviation was evaluated.Results The background counting was 0×109/L and the carryover rate was 0.1%,conformed to the quality requirements of hematology analyzer,and the repeatability study imprecision ranged between 4.7%-18.8%.HPC of peripheral venous blood linear range(0-27.201×109/L),Stem cell collection was diluted 5 times linear range(0-0.878×109/L).The results of 126 samples detected by the hematology analyzer and FCM were compared.The correlation coefficient r2=0.9601.When WBC>10×109/L,the results of the two instruments have a good consistency.The slope is between 0.95 and 1.05,and the relative bias is less than 30%.Conclusions This study suggests that the hematology analyzer has a good linear range for detecting HPC,and has a good correlation with FCM.The hematology analyzer has the advantages of no pretreatment,convenient operation,a wide range of applications in detecting HPC specimens.