The Role of Volume, Conductivity, Scatter Changes of Neutrophils and Monocytes in Diagnosis of Megaloblastic Anemia

  • Deepa Sowkur Anandarama Adiga Department of Pathology Kasturba Medical College, Mangalore- 01 MAHE, Manipal, Karnataka, India
  • Debarshi Saha Department of Pathology Kasturba Medical College, Mangalore- 01 MAHE, Manipal, Karnataka, India
  • Karthick R G Clinical Hematology Resident Narayana Hrudayalaya, Bangalore
  • Vishnu Priya M Department of Pathology Fellow in Clinical Hematology HCG, Bangalore
  • Purnima S Rao Department of Pathology A J Institute of Medical Sciences Mangalore-01
  • Pooja K Suresh Department of Pathology Kasturba Medical College, Mangalore- 01 MAHE, Manipal, Karnataka, India
Keywords: Megaloblastic anemia, automated hematology analyzer, neutrophils, monocytes


Background: Megaloblastic anemias are macrocytic normochromic anemia with mean corpuscular volume (MCV) of 100 fl-140 fl and caused by deficiency of either cobalamin (vitamin B12) or folate. However, increased MCV is not specific for megaloblastic anemia, nor is Vitamin B12 assay by chemiluminescence. We undertook this study to evaluate the possible role of Volume, Conductivity and Scatter (VCS) of WBCs derived from standard hematology analyzer to indicate megaloblastic anemia. Methods: We performed a case control study comparing data of 60 patients with low serum vitamin B12 or folate levels with 60 healthy volunteers. Comparison of the volume, conductivity and scatter parameters for neutrophils and monocytes of cases and control were done. Result: The mean neutrophil volume of cases (158.37±18.13fl) was significantly higher (p= 0.0001) compared to controls (141.26±4.22fl). Similarly, mean monocyte volume of cases (183.34±16.90fl) was significantly (p=0.0001) higher compared to controls (166.55±8.66fl). The difference in the mean conductivity of both neutrophils and monocytes between cases and controls were insignificant (p=0.43). Conclusion: Our study suggests analysis of VCS parameters for neutrophils and monocytes was a simple and objective method that substantiates the existence of subclinical deficiency of vitamin B 12 and folate with fair degree of certainty.


Hoffbrand AV. 2013. Megaloblastic anemia. In: Longo DL, editor. Harrison’s hematology and oncology 17th edition. New York: McGraw-Hill Medical. p 94-108.

McKenzie SB, Williams L. 2014. Megaloblastic and non megaloblastic macrocytic anemias. In: McKenzie SB, Williams L, editors. Clinical Laboratory Hematology 3rdedition. Essex: Pearson. p 311-324.

Bhatia P, Kulkarni JD, Pai SA.Vitamin B12 deficiency in India:Mean Corpuscular Volume is an unreliable screening parameter. Nat Med J India 2012; 25: 336-8

Mardi D, Fwity B, Lobmann R, Ambrosch A. Mean cell volume of neutrophils and monocytes compared with C-reactive protein,IL-6 and white blood cell count for prediction of sepsis and non systemic bacterial infections. Int J Lab Hematol 2010; 32: 410-18.

Chaves F, Tierno B, Xu D. Quantitative Determination of Neutrophil VCS Parameters by the Coulter Automated Hematology Analyzer. Am J Clin Pathol. 2005; 124: 440-44.

Aslinia F, Mazza JJ, Yale SH Megaloblastic anemia and other causes of macrocytosis. Clin Med Res 2006; 4: 236–241,

Kaferle J, Strzoda CE (2009) Evaluation of macrocytosis. Am Fam Physician 79(3):203–208

Ganji V, Kafai MR. Trends in prevalence of megaloblatic anemia in US adults: comparative analysis of pre- and post-folic acid fortification surveys.

Carmel R., Green R., Jacobsen D.W. & Qian G.D. (1996) Neutrophil nuclear segmentation in mild cobalamin deficiency metabolic tests of cobalamin status and observations on ethnic differences in neutrophil segmentation. American Journal of Clinical Pathology 106, 57–63

Oosterhuis WP, Niessen RWLM, Bossuyt PMM, Sanders GTB, Sturk A. Diagnostic value of the mean corpuscular volume in the detection of vit B12 deficiency.Scand J Clin Lab Invest 2000;60:9-18.

Hawaldar R, Sodani S. Study of BECKMAN COULTER LH 750 CPD parameters in subclinical Vitamin B12 deficiency. Ind J Pathol Oncol. 2016; 3(4); 704-9.

Vlasveld LT, Van’t Wout JW, Meeuwissen P, Castel A. High measured cobalamin (vitamin B12) concentration attributable to an analytical problem in testing serum from a patient with pernicious anemia. Clin Chem 2006; 52:157–8.

Hamilton MS, Blackmore S, Lee A. Possible cause of false normal B-12 assays. BMJ 2006; 333:654–5.

Bailey RL, Carmel R, Green R, Pfeiffer CM, Cogswell ME, Osterloh JD, Sempos CT, Yetley EA. Monitoring of vitamin B12 nutritional status in the Unites States by using plasma methylmalonic acid and serum vitamin B12. Am J Clin Nutr 2011;94:552–61

Antony AC. Megaloblastic anemia. In: Hematology: Basic Principles and Practice, 4th edn. Hoffman R, Benz EJ, Shattil SJ, Furie B, Cohen HJ, Silberstein LE, McGlave P. (eds). New York: Churchil Livingstone, 2005: 519–57.

Allen RH, Stabler SP, Savage DG, Lindenbaum J. Diagnosis of cobalamin deficiency I: usefulness of serum methylmalonic acid and total homocysteine concentrations. Am J Hematol 1990; 34: 90–8.

Lindenbaum J, Savage DG, Stabler SP, Allen RH. Diagnosis of cobalamin deficiency: II. Relative sensitivities of serumcobalamin, methylmalonic acid, and total homocysteine concentrations. Am J Hematol 1990; 34: 99–107.].

Chaves F, Tierno B, Xu D Neutrophil volume distribution width: a new automated hematologic parameter for acute infection. Arc Pathol Lab Med 2006; 130: 378–380

Park DH, Park K, Park J, Park HH, Chae H, Lim J, Oh EJ, Kim Y, Park YJ, Han K. Screening of sepsis using leukocyte cell population data from the Coulter automatic blood cell analyzer DxH800. Int J Lab Hematol 2011; 33: 391–399

Raimondi F, Ferrara T, Capasso L, Sellitto M, Landolfo F, Romano A, Grimaldi E, Scopacasa F. Automated determination of neutrophil volume as screening test for late-onset sepsis in very low birth infants. Pediatr Inf Dis J 2011; 29: 288

Risch C, Medina P, Nydegger UE, Bahador Z, Brinkmann T, Landenberg PV. Relationship of Leukocyte anisocytosis to holotranscobalamin, a marker for cobalamin deficiency. Int J Lab Haematol. 2012; 34: 192-200.

Totoskovic D, Dopsaj V, Martinovic J. Methylmalonic acid and neutrophil morphometric index in laboratory diagnosis of cobalamin deficiency without macrocytosis. Int J Lab Hematol 2016, 38, 265–272

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