• 四川大學(xué)華西醫(yī)院（成都，610041）1干細(xì)胞生物學(xué)研究室，2實驗醫(yī)學(xué)科;

目的 　建立急性白血?。ˋL）患者八色流式免疫表型分析起始管方案。 方法 　用胞膜CD3（CD3）、CD19、CD10、CD34、CD45、胞漿CD79a（cCD79a）、髓過氧化物酶（MPO)和胞漿CD3（cCD3）等8種抗體建立八色流式染色方案。膜表面抗體直接染色；膜內(nèi)抗體經(jīng)固定破膜，再染色后上機(jī)檢測。將3個血小板減少患者骨髓標(biāo)本分別進(jìn)行抗體的單色染色和缺一色染色；最后對17例確診的AL初發(fā)患者標(biāo)本進(jìn)行檢測。 結(jié)果　 用單色染色來確定染色方案中各抗體的檢測電壓及熒光補(bǔ)償；缺一色染色中，陽性細(xì)胞群較單色染色變化均＜10%，表明方案中的各抗體相互作用小。17例AL初發(fā)患者中，6例急性B淋巴細(xì)胞白血病原始細(xì)胞均為CD34和CD19陽性，5例cCD79a陽性和4例CD10陽性；4例急性T淋巴細(xì)胞白血病患者均為cCD3陽性；6例急性髓細(xì)胞白血病均為CD34和MPO陽性；1例B+T混合表型AL患者CD34、cCD3、CD19、cCD79a及CD10均為陽性，MPO和CD3為陰性，此檢測方案能夠確定各類AL的細(xì)胞類型。 結(jié)論 　建立了AL患者八色流式免疫表型分析起始管方案，操作簡便快速，適用于臨床檢測。

引用本文： 黃巧容,余建華,蔣能剛,曾婷婷,賈永前,孟文彤. 急性白血病患者八色流式免疫表型分析起始管方案的建立. 華西醫(yī)學(xué), 2012, 27(4): 555-559. doi: 復(fù)制

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6. 　Craig FE, Foon KA. Flow cytometric immunophenotyping for hematologic neoplasms[J]. Blood, 2008, 111(8): 3941-3967.
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12. 　李麗, 付笑迎, 吳長有. 多色流式細(xì)胞術(shù)檢測人外周血BCG特異性效應(yīng)型記憶γδＴ特異性效應(yīng)型記憶[J]. 免疫學(xué)雜志, 2011, 27(5): 369-372.
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14. 　Chattopadhyay PK, Hogerkorp CM, Roederer M. A chromatic explosion: the development and future of multiparameter flow cytometry[J]. Immunology, 2008, 125(4): 441-449.
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  2. 2. 　牛挺, 劉霆. 造血干細(xì)胞移植并發(fā)癥的防治進(jìn)展[J]. 華西醫(yī)學(xué), 2011, 26 (12): 1761-1762.
  3. 3. 　Brüggemann M, Schrauder A, Raff T, et al. Standardized MRD quantification in European ALL trials: proceedings of the second international symposium on MRD assessment in Kiel, Germany, 18–20 September 2008[J]. Leukemia, 2010, 24 (3): 521-535.
  4. 4. 　Pedreira CE, Costa ES, Barrena S, et al. Generation of flow cytometry data files with a potentially infinite number of dimensions[J]. Cytometry A, 2008, 73 (9): 834-846.
  5. 5. 　Weinberg OK, Arber DA. Mixed-phenotype acute leukemia: historical overview and a new definition[J]. Leukemia, 2010, 24(11): 1844-1851.
  6. 6. 　Craig FE, Foon KA. Flow cytometric immunophenotyping for hematologic neoplasms[J]. Blood, 2008, 111(8): 3941-3967.
  7. 7. 　Matutes E, Pickl WF, van’t VeerM, et al. Mixed-phenotype acute leukemia: clinical and laboratory features and classification outcome in 100 patients defined according to the WHO 2008[J]. Blood, 2011, 117(11): 3163-3171.
  8. 8. 　van den Ancker W, Terwijn M, Westers TM, et al. Acute leukemias of ambiguous lineage: diagnostic consequences of the WHO 2008 classification [J]. Leukemia, 2010, 24(7): 1392-1396.
  9. 9. 　Wood B. 9-color and 10-color flow cytometry in the clinical laboratory[J]. Arch Pathol Lab Med, 2006, 130(5): 680-690.
  10. 10. 　Preffer F, Dombkowski D. Advances in complex multiparameter flow cytometry technology: applications in stem cell research[J]. Cytometry B Clin Cytom, 2009, 76(5): 295-314.
  11. 11. 　李麗, 付笑迎, 吳長有. 八色流式細(xì)胞術(shù)檢測人外周血BCG特異性效應(yīng)型記憶CD4~+T細(xì)胞的表型特征[J]. 細(xì)胞與分子免疫學(xué)雜志，2011, 27(5): 491-493.
  12. 12. 　李麗, 付笑迎, 吳長有. 多色流式細(xì)胞術(shù)檢測人外周血BCG特異性效應(yīng)型記憶γδＴ特異性效應(yīng)型記憶[J]. 免疫學(xué)雜志, 2011, 27(5): 369-372.
  13. 13. 　Mahnke YD, Roederer M. Optimizing a multicolor immunophenotyping assay[J]. Clin Lab Med, 2007, 27(3): 469-487.
  14. 14. 　Chattopadhyay PK, Hogerkorp CM, Roederer M. A chromatic explosion: the development and future of multiparameter flow cytometry[J]. Immunology, 2008, 125(4): 441-449.