Blood Res.  2014 Jun;49(2):120-126. 10.5045/br.2014.49.2.120.

White blood cell differential counts in severely leukopenic samples: a comparative analysis of different solutions available in modern laboratory hematology

• Kim AH
• Lee W
• Kim M
• Kim Y
• Han K

• KMID: 2172808
• DOI: http://doi.org/10.5045/br.2014.49.2.120

Abstract

BACKGROUND
We evaluated the efficacy of white blood cell (WBC) differential counts in severely leukopenic samples by the Hematoflow method and by automated hematology analyzers and compared the results with manual counts.
METHODS
EDTA-anticoagulated blood samples (175 samples) with WBC counts of 40-990/microL were selected. Hematoflow differential counts were performed in duplicates employing flow cytometry using the CytoDiff reagent and analysis software. Differential counts were also performed using the DxH 800 (Beckman Coulter) and XE-2100 (Sysmex) automated hematology analyzers. The sum of the manual counts by a hematology technician and a resident were used as the manual counts.
RESULTS
The total analysis time and hands-on time required by the Hematoflow method were shorter than those required by manual counting. Hematoflow counts were reproducible, showed a good correlation with automated analyzers, and also showed strong correlation with manual counts (r > 0.8) in neutrophils, lymphocytes, and monocytes. None of the cases containing less than 4% blasts as analyzed by the Hematoflow method had blasts in the manual counts, but 8 cases of 21 cases (38.1%) with over 4% blasts by Hematoflow had blasts in manual counts.
CONCLUSION
Hematoflow counts of severely leukopenic samples were reproducible and showed a good correlation with manual counts in terms of neutrophil, lymphocyte, and monocyte counts. The Hematoflow method also detected the presence of blasts. Manual slide review is recommended when over 4% blasts are found by Hematoflow.

Keyword

Flow cytometry; Leukopenia; Differential leukocyte count

MeSH Terms

Flow Cytometry
Hematology*
Leukocyte Count
Leukocytes*
Leukopenia
Lymphocytes
Monocytes
Neutrophils

Figure

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  Fig. 1 An example of Hematoflow results. Seventeen cell populations are displayed in different colors using specific gates recommended by the manufacturer.

Reference

1. Mansberg HP, Saunders AM, Groner W. The Hemalog D white cell differential system. J Histochem Cytochem. 1974; 22:711–724. PMID: 4137312.

2. Siekmeier R, Bierlich A, Jaross W. The white blood cell differential: three methods compared. Clin Chem Lab Med. 2001; 39:432–445. PMID: 11434394.

3. Guerti K, Vertessen F, Daniels L, Van Der Planken M. Performance evaluation of the PENTRA 60C+ automated hematology analyzer and comparison with the ADVIA 2120. Int J Lab Hematol. 2009; 31:132–141. PMID: 19267810.

4. Novis DA, Walsh M, Wilkinson D, St Louis M, Ben-Ezra J. Laboratory productivity and the rate of manual peripheral blood smear review: a College of American Pathologists Q-Probes study of 95,141 complete blood count determinations performed in 263 institutions. Arch Pathol Lab Med. 2006; 130:596–601. PMID: 16683868.

5. Barnes PW, McFadden SL, Machin SJ, Simson E. The international consensus group for hematology review: suggested criteria for action following automated CBC and WBC differential analysis. Lab Hematol. 2005; 11:83–90. PMID: 16024331.

6. Burchert-Graeve M, Kock R. Automated leucocyte differentials in 292 patients with leucopenia: an evaluation of the Abbott CELL-DYN 3500 (CD3500) haematology analyser. Clin Lab Haematol. 1996; 18:253–259. PMID: 9054698.

7. Saloustros E, Tryfonidis K, Georgoulias V. Prophylactic and therapeutic strategies in chemotherapy-induced neutropenia. Expert Opin Pharmacother. 2011; 12:851–863. PMID: 21254862.

8. Kulkarni S, Ghosh SP, Hauer-Jensen M, Kumar KS. Hematological targets of radiation damage. Curr Drug Targets. 2010; 11:1375–1385. PMID: 20583980.

9. Fuentes-Arderiu X, Garcia-Panyella M, Dot-Bach D. Between-examiner reproducibility in manual differential leukocyte counting. Accred Qual Assur. 2007; 12:643–645.

10. Pierre RV. Peripheral blood film review. The demise of the eyecount leukocyte differential. Clin Lab Med. 2002; 22:279–297. PMID: 11933579.

11. Faucher JL, Lacronique-Gazaille C, Frebet E, et al. "6 markers/5 colors" extended white blood cell differential by flow cytometry. Cytometry A. 2007; 71:934–944. PMID: 17879238.

12. Jo Y, Kim SH, Koh K, et al. Reliable, accurate determination of the leukocyte differential of leukopenic samples by using Hematoflow method. Korean J Lab Med. 2011; 31:131–137. PMID: 21779183.

13. Elliott MA, Litzow MR, Letendre LL, et al. Early peripheral blood blast clearance during induction chemotherapy for acute myeloid leukemia predicts superior relapse-free survival. Blood. 2007; 110:4172–4174. PMID: 17909077.

14. Manabe A, Ohara A, Hasegawa D, et al. Significance of the complete clearance of peripheral blasts after 7 days of prednisolone treatment in children with acute lymphoblastic leukemia: the Tokyo Children's Cancer Study Group Study L99-15. Haematologica. 2008; 93:1155–1160. PMID: 18519521.

15. Segal BH, Freifeld AG. Antibacterial prophylaxis in patients with neutropenia. J Natl Compr Canc Netw. 2007; 5:235–242. PMID: 17335692.

16. Shafer JA. Blood and marrow morphology in acute leukemia patients receiving chemotherapy: a photo-essay. Am J Med Technol. 1983; 49:77–90. PMID: 6573133.

17. Simson E, Gascon-Lema MG, Brown DL. Performance of automated slidemakers and stainers in a working laboratory environment - routine operation and quality control. Int J Lab Hematol. 2010; 32:e64–e76. PMID: 19220552.

18. DiNardo CD, Luger SM. Beyond morphology: minimal residual disease detection in acute myeloid leukemia. Curr Opin Hematol. 2012; 19:82–88. PMID: 22314322.

19. Patkar N, Alex AA, B B, et al. Standardizing minimal residual disease by flow cytometry for precursor B lineage acute lymphoblastic leukemia in a developing country. Cytometry B Clin Cytom. 2012; 82:252–258. PMID: 22467604.

20. van de Geijn GJ, van Rees V, van Pul-Bom N, et al. Leukoflow: multiparameter extended white blood cell differentiation for routine analysis by flow cytometry. Cytometry A. 2011; 79:694–706. PMID: 21786418.

21. Jean A, Boutet C, Lenormand B, et al. Combination of cellular population data and CytoDiff analyses for the diagnosis of lymphocytosis. Clin Chem Lab Med. 2011; 49:1861–1868. PMID: 21875393.

22. de Tute RM. Flow cytometry and its use in the diagnosis and management of mature lymphoid malignancies. Histopathology. 2011; 58:90–105. PMID: 21261686.