单管10色流式细胞术分类计数外周血白细胞

目的 应用"单管10色"流式细胞术分类计数外周血白细胞,并探讨其临床应用价值.方法 通过多种单克隆抗体的选择和多重逻辑设门的研究,确定了10色流式细胞术与12种单克隆抗体组合的单管免洗外周血白细胞分类方法(简称"单管10色"方法).选取142份外周血标本,以血涂片显微镜法分类技术为参考方法,同时用Beckman Cotdter LH750全自动血细胞分析仪和"单管10色"方法分类计数白细胞,计算"单管10色"方法的临床诊断效率,并统计分析两种方法与显微镜法分类计数白细胞的相关性.以BD FACS Calibur流式细胞仪免疫分型结果为标准,计算"单管10色"流式细胞术检出原始细胞的诊断效率.结果 "单管10色"流式细胞术与显微镜法分类计数外周血白细胞除嗜碱粒细胞外均具有良好的相关性(r均>0.700,P均<0.01),其中对检出中性粒细胞、淋巴细胞、未成熟粒细胞和原始细胞的相关性较高(r＝0.972、0.951、0.801、0.912,P均<0.01).以显微镜法分类计数外周血未成熟粒细胞1%为临界值,"单管10色"流式细胞术检出未成熟粒细胞敏感度为92%(57/62),特异度为79%(63/80).以显微镜法检出外周血原始细胞0.5%为临界值,"单管10色"流式细胞术检出原始细胞敏感度为99%(67/68),特异度为92%(68/74).以BD FACSCalibur流式细胞仪免疫分型结果为标准,"单管10色"流式细胞术对原始细胞检出的敏感度为100%(40/40),特异度为91%(10/11),阳性预测值为98%(40/41),阴性预测值为100%(10/10),准确性为98%(50/51).结论 "单管10色"流式细胞术分类计数外周血白细胞与显微镜法具有良好的相关性,有望用于全血细胞计数异常时的后续检验.

Abstract：

Objective To explore the values of potential clinical application of"single tube/ten colors"flow cytometry for leukocyte differential count in peripheral blood.Methods Utilizing multiple monoclonal antibody combinations and the vavious logical gating strategies,the single tube/12 antibodies with no-wash method for the leukocyte differential count in peripheral blood were determined by using 10 colors flow cytometry.Leukocyte differentials of 142 peripheral blood samples were determined by both Beckman-Coulter LH750 hematology analyzer and 10 colors flow cytometry.The results were then compared to standard microscopic examination as a reference method.The clinical diagnostic efficiency of"single tube/10 colors"flow cytometry was calculated.The correlations between standard microscopic cytology,"single tube/10 colors"flow cytometry and the hematology analyzer were determined.In addition,the clinical diagnosis efficiency for blast counts of"single tube/10 colors"were compared to the results determined by BD FACS Calibur flow cytometer.Results The leukocyte differentials were correlated well between the "single tube/10 colors"flow cytometry and standard microscopic cytology(r>0.700,P<0.01) except for basophils.The correlations with neutrophilic granulocytes,lymphocytes,immature granulocytes and blasts were superior(r=0.972,0.951,0.801,0.912,respectively,P<0.01).When 1% was selected as the cut-off point for immature granulocytes determined by standard microscopic cytology,the sensitivity and the specificity of"single tube/10 colors"flow cytometry were 92%(57/62) and 79% (63/80),respectively.When 0.5% was selected as the cut-off point for blasts detected by standard microscopic cytology,the sensitivity and the specificity were 99% (67/68) and 92% (68/74).Using the immunophenotyping results from BD FACS Calibur as a standard,the sensitivity for detecting blasts by"single tube/10 colOrs"flow cytometry was 100% (40/40),the specificity was 91% (10/11),the positive predictive value was 98% (40/41),the negative predictive value was 100% (10/10) and the accuracy was 98% (50/51).Conclusions The"single tube/10 colors"flow cytometry has a excellent correlation with the standard microscopic cytology when applied on leukocyte differential count in peripheral blood.It may potentially use as a subsequent method for verification of abnormal results of complete blood cell count in the future.

Leukocyte differential count in peripheral blood "with" single tube/ten colors"flow cytometry

Objective To explore the values of potential clinical application of"single tube/ten colors"flow cytometry for leukocyte differential count in peripheral blood.Methods Utilizing multiple monoclonal antibody combinations and the vavious logical gating strategies,the single tube/12 antibodies with no-wash method for the leukocyte differential count in peripheral blood were determined by using 10 colors flow cytometry.Leukocyte differentials of 142 peripheral blood samples were determined by both Beckman-Coulter LH750 hematology analyzer and 10 colors flow cytometry.The results were then compared to standard microscopic examination as a reference method.The clinical diagnostic efficiency of"single tube/10 colors"flow cytometry was calculated.The correlations between standard microscopic cytology,"single tube/10 colors"flow cytometry and the hematology analyzer were determined.In addition,the clinical diagnosis efficiency for blast counts of"single tube/10 colors"were compared to the results determined by BD FACS Calibur flow cytometer.Results The leukocyte differentials were correlated well between the "single tube/10 colors"flow cytometry and standard microscopic cytology(r>0.700,P<0.01) except for basophils.The correlations with neutrophilic granulocytes,lymphocytes,immature granulocytes and blasts were superior(r=0.972,0.951,0.801,0.912,respectively,P<0.01).When 1% was selected as the cut-off point for immature granulocytes determined by standard microscopic cytology,the sensitivity and the specificity of"single tube/10 colors"flow cytometry were 92%(57/62) and 79% (63/80),respectively.When 0.5% was selected as the cut-off point for blasts detected by standard microscopic cytology,the sensitivity and the specificity were 99% (67/68) and 92% (68/74).Using the immunophenotyping results from BD FACS Calibur as a standard,the sensitivity for detecting blasts by"single tube/10 colOrs"flow cytometry was 100% (40/40),the specificity was 91% (10/11),the positive predictive value was 98% (40/41),the negative predictive value was 100% (10/10) and the accuracy was 98% (50/51).Conclusions The"single tube/10 colors"flow cytometry has a excellent correlation with the standard microscopic cytology when applied on leukocyte differential count in peripheral blood.It may potentially use as a subsequent method for verification of abnormal results of complete blood cell count in the future.