[1]方佩琪,孙 林,顾梅秀,等.碧迪FACS CantoⅡ流式细胞仪性能评价及实验室应用[J].现代检验医学杂志,2019,34(02):94-99.[doi:10.3969/j.issn.1671-7414.2019.02.025]
　FANG Pei-qi,SUN Lin,GU Mei-xiu,et al.Evaluation and Clinical Application of BD FACS Canto II Flow Cytometer[J].Journal of Modern Laboratory Medicine,2019,34(02):94-99.[doi:10.3969/j.issn.1671-7414.2019.02.025]
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碧迪FACS CantoⅡ流式细胞仪性能评价及实验室应用()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第34卷
期数:
2019年02期
页码:
94-99
栏目:
研究简报·实验技术
出版日期:
2019-05-11

文章信息/Info

Title:
Evaluation and Clinical Application of BD FACS Canto II Flow Cytometer
文章编号:
1671-7414(2019)02-094-06
作者:
方佩琪; 孙 林; 顾梅秀; 吴 蕙; 潘柏申; 郭 玮; 王蓓丽
(复旦大学附属中山医院检验科,上海 200032)
Author(s):
FANG Pei-qi; SUN Lin; GU Mei-xiu; WU Hui; PAN Bai-shen; GUO Wei; WANG Bei-li
(Department of Laboratory Medicine,Zhongshan Hospital of Fudan University,Shanghai 200032,China)
关键词:
BD FACS CantoⅡ;  流式细胞仪;  性能评价
分类号:
R446
DOI:
10.3969/j.issn.1671-7414.2019.02.025
文献标志码:
A
摘要:
目的 评估碧迪FACS CantoⅡ(BD FACS CantoⅡ)流式细胞仪的性能及实验室应用。方法 依据中华人民共和国医药行业新版标准《YY/T0588-2017流式细胞仪》[1]对BD FACS CantoⅡ流式细胞仪进行性能评价,评价内容包括:荧光检出限、荧光线性、前向角散射光(forward scatter resolution,FSC)检出限、仪器分辨率、FSC分辨率、侧向角散射光(side scattering resolution,SSC)分辨率、倍体分析线性、表面标志物检测的准确度、表面标志物检测的重复性、携带污染率和仪器稳定性。完成性能评价后,分别用BD FACS CantoⅡ流式细胞仪及其配套的BD六色淋巴细胞亚群试剂盒和BD FACS Calibur流式细胞仪及其配套的BD四色淋巴细胞亚群试剂盒分别检测20份全血标本,比较两台仪器在标本前处理、上机检测、分析结果所用时间和结果比较的差异。结果 该流式细胞仪对异硫氰酸荧光素(fluorescein isothiocyanate,FITC)通道荧光检出限为40.18等量可溶性荧光分子数(molecules equiva-lent soluble fluorochrome,MESF),对藻红蛋白(P-phycoerythrin,PE)通道荧光检出限为23.77 MESF,两者的决定系数(R square,r)均为0.999 9; FSC最小可检测0.22 μm微球; 仪器分辨率FSC,FITC和PE荧光通道的变异系数(coeffivirnt of cariation,CV)分别为2.3%,2.4%和2.4%; FSC/SSC散点图可明显区分外周血中红细胞和血小板,同时能区分白细胞中的淋巴细胞、中性粒细胞、单核细胞; 倍体分析线性中G2/M期(四倍体细胞峰)与G0/G1期(二倍体细胞峰)的平均荧光强度比值为1.98; 表面标志物检测值的平均值均在质控品说明书给定的靶值参考范围内; 表面标志物检测重复性CV分别为CD3:2.75%,CD4:1.86%,CD8:5.14%,CD16/CD56:7.33%和CD19:11.26%; 携带污染率最大为0.05%; 该仪器开机8 h检测FSC及各荧光通道峰值荧光道数偏差值(B)分别为-0.40%,-0.55%,1.57%,1.88%,1.66%,7.44%,7.23%,6.63%和6.21%; BD FACS Canto Ⅱ流式细胞仪及其配套试剂盒较BD FACS Calibur流式细胞仪及其配套试剂盒在总体检测速度上有显著提高; 两台仪器所得的检测结果差异无统计学意义(P>0.05),且前者检测结果的离散程度小于后者。结论 BD FACS Canto Ⅱ流式细胞仪各项性能均符合行业标准,仪器各项技术指标达标,可有效保障实验室检测结果的准确性,能为临床诊疗提供可靠支持。
Abstract:
Objective To evaluate the performance of BD FACS CantoII flow cytometer.Methods According to the latestmedicine industry standard of the People's Republic of China 《YY/T0588-2017flow cytometry》[1],evaluated the performance of flow cytometry such asfluorescence sensitivity,fluorescence linearity,forward scatter sensitivity(FSC),instrument resolution,forward scatter/side scattering resolution(SSC),DNAcontent linearity,carry-over rate,accuracy of the cell surface marker,reproducibility of the cell surface marker,instrument stability and clinical application.After the evaluation,used the BD FACS Canto Ⅱ and supporting the BD MultitestTM 6-color TBNK Reagent and BD FACS Calibur flow cytometer and supporting the BD MultitestTM IMK Kit,than testing 20 cases of whole blood samples.Results The fluorescence detection limit was 40.18molecules equiva-lent soluble fluorochrome(MESF)of fluorescein isothiocyanate(FITC)channel and 23.77 MESF of P-phycoerythrin(PE)channel,both r values were 0.999 9.The detection limit for FSC could detect 0.22 μm microspheres,instrument resolution:coeffivirnt of cariation(CV)of FSC,FITE,PE channel were 2.3%,2.4% and 2.4%.The scatter plots of FSC/SSC could distinguish thyrocyte,platelets in peripheral blood,and lymphocytes,neutrophils and monocytesin leukocyte.The average fluorescence intensity ratio of DNA content linearityG0/G1 and G2/M was 1.98.The average value of all surface markers was within the reference range.The coefficient of variation of surface marker detection repeatability was CD3:2.75%,CD4:1.86%,CD8:5.14%,CD16/CD56:7.33% and CD19:11.26%,and the carry-overrate up to 0.05%.Instrument stability,the deviation(B)of each fluorescent channel after 8 hours of starting was:-0.40%,-0.55%,1.57%,1.88%,1.66%,7.44%,7.23%,6.63% and 6.21%,respectively.Validation results related to clinical applications showed its advantages in effectively improving productivity.The BD FACS Canto II flow cytometer and its supporting kit had significantly improved the testing speed compared to the BD FACS Calibur flow cytometer and its supporting kit.The difference between the two system was not statistically significant(P>0.05),and the former was less discrete than the latter.Conclusion The performance of BD FACS Canto II flow cytometer met the requirements of industry standard.The system value were up to the standard,and guarantee the accuracy of laboratory results,also are reliablesupport for clinical medical.

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备注/Memo

备注/Memo:
基金项目:上海市卫生计生系统重要薄弱学科建设项目(2015ZB0201); 国家自然科学基金(81572064,81772263); 上海市卫生和计划生育委员会科研课题计划任务书(201440389); 上海市科学技术委员会(16411952100)。 作者简介:方佩琪(1988-),女,本科,技师,主要从事血液及体液流式细胞术检测,E-mail:fang.peiqi@zs-hospital.sh.cn。 通讯作者:王蓓丽,女,博士,副主任技师,E-mail:wang.beili1@zs-hospital
更新日期/Last Update: 2019-04-30