• 四川大學華西醫(yī)院(成都,610041)1干細胞生物學研究室,2實驗醫(yī)學科;

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,再染色后上機檢測。將3個血小板減少患者骨髓標本分別進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)患者標本進行檢測。 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償;缺一色染色中,陽性細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淋巴細胞白血病原始細胞均為CD34和CD19陽性,5例cCD79a陽性和4例CD10陽性;4例急性T淋巴細胞白血病患者均為cCD3陽性;6例急性髓細胞白血病均為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, 2012, 27(4): 555-559. doi: 復(fù)制

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  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. 八色流式細胞術(shù)檢測人外周血BCG特異性效應(yīng)型記憶CD4~+T細胞的表型特征[J]. 細胞與分子免疫學雜志,2011, 27(5): 491-493.
  12. 12.  李麗, 付笑迎, 吳長有. 多色流式細胞術(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.