DOI: http://dx.doi.org/10.18203/2320-6012.ijrms20194317

Comparison of automated flowcytometric reticulocyte analysis with manual reticulocyte count

Komal Patel, Siddhi M. Patel

Abstract


Background: Reticulocytes are young or immature red blood cells released from bone marrow and that contain remanants of ribonucleic acid (RNA) and ribosomes. Reticulocyte count (RC) is the index of erythropoietic activity within bone marrow. The reticulocyte counting methods at clinical laboratories are currently divided into manual and automated.

Methods: A total of 500 samples of study cases were processed by manual method using New Methylene Blue (NMB) and automated method based on flowcytometry by PENTRA XLR HORIBA hematology analyzer. All quality control parameters were evaluated and values obtained by both methods were compared using various statistical methods.

Results: Automated hematology analyzer provides excellent precision and linearity with no significant carryover. On comparing manual and automated RC method good method correlation was found (correlation coefficient r-0.865), however individual case wise percent deviation between manual and automated RC and CRC varied significantly. In addition within run precision calculated for automated RC differed significantly from manual count. The mean of difference between duplicate readings (150 samples) of manual and automated RC (<5%) were 0.3 and 0.01 respectively while 6.3 and 0.15 respectively for >5% RC. Thus, automated method was found to be more precise than the manual RC.

Conclusions: The manual count method for RC associated with significant imprecision compared to flowcytometric method mostly based on interobserver variation and the smaller number of cell being counted. In contrast, the automated method is rapid, easy to operate, count higher number of cells with precise measurement.


Keywords


Automated method, PENTRA XLR( HORIBA) hematology analyzer, Manual method, Reticulocyte count

Full Text:

PDF

References


Singh A, Rastogi S, Garg DK, Singh D, Ahmad K, Chhabra P. Automated reticulocyte count wins over manual methods. J Med Sci Clin Research. April 2016; 4(4):10231-3.

Riley RS, Ben-Ezra JM, Tidwell A. Reticulocyte enumeration: past & present. Laboratory Med. 2001 Oct 1;32(10):599-608.

Rastogi S, Singh A, Chhabra P. Automated Corrected Reticulocyte Count Superiority above manual methods. Scholars J Appl Medi Scien 2016;4(4A):1177-9.

Cortellazzi LC, Teixeira SM, Borba R, Gervásio S, Cintra CS, Helena ZW. Reticulocyte parameters in hemoglobinopathies and iron deficiency anemia; Rev. bras. hematol. hemator. 2003;25(2):97-102.

Buttarello M. Laboratory diagnosis of anemia: are the old and new red cell parameters useful in classification and treatment, how?; Indian J Labor hematol. 2016;38(1):123-32.

Raja-Sabudin RZ, Othman A, Ahmed-Mohamed KA, Ithnin A, Alauddin H, Alias H, et al. Immature reticulocyte fraction is an early predictor of bone marrow recovery post chemotherapy in patients with acute leukemia. Saudi Med J. 2014 Apr 1;35(4):346-9.

Viana KA, Filho O, Sant’Ana Dusse L, Avelar RS, Vitelli Avelar D, Carvalho B, Franco Ribeiro C, et al. Reticulocyte count: comparison among methods. J Bras Med Lab. 2014;50(5):339-5.

Rowan RM. Reference method, quality control and automation of reticulocyte count. Pure and applied chemistry. 1991;63(8):1141-5.

Buttarello M, Bulian P, Farina G, Temporin V, Toffolo L, Trabuio E, et al. Flow Cytometric Reticulocyte Counting: Parallel Evaluation of Five Fully Automated Analyzers: An NCCLS-ICSH Approach, Am J Clil Path. 1 January 2001;115(1): 100-11.

Koepke JA. Update on Reticulocyte Counting. Laboratory medicine. May 1999; 30(5): 339-43.

Davis BH. Immature reticulocyte fraction (IRF): by any name, a useful clinical parameter of erythropoietic activity. Lab Hematol. 1996;2:2-8.

Davis BH, Bigelow NC, Koepke JA, Borowitz MJ, Houwen B, Jacobberger JW, et al. Flow cytometric reticulocyte analysis: multiinstitutional interlaboratory correlation study. Am J Cli Pat. 1994 Oct 1;102(4):468-77.

Davis BH, Ornvold K, Bigelow NC. Flow cytometric reticulocyte maturity index: a useful laboratory parameter of erythropoietic activity in anemia. Cytometry: J Inte Soc Analy Cytol. 1995 Mar 15;22(1):35-9.

Piva E, Brugnara C. Clinical utility of reticulocyte parameters. Clinics in laboratory medicine 2015; 35(1):133-63.

Smith T. On changes in the red blood corpuscles in pernicious anemia of Texas cattle fever. Trans Assoc Am Physicians. 1891;6:263-77.

Szigeti RG. Reticulocyte Count and Reticulocyte Hemoglobin Content. Sep 05, 2014.

Toruntju SA, Arif M , Hadju V, Syam A, Hamid SK, Thaha AR. Description of Reticulocyte-Hemoglobin, Immature Reticulocyte Fraction and Reticulocyte Production Index on the Student 12-15 Years Who Get Local Fortification Rice. Int J heath science and research. 2017;7(4):40-6.

HORIBA. User manual for PENTRA XLR Hematol analyzer:1-318.

Lacombe F, Lacoste L, Vial JP, Briasis A, Josy R, Boisseau MR, et al. Comparison of ABX PENTRA 120 Retic, Sysmex R-2000, Flow Cytometry and manual counts. Am J Clin Pathol. 1999;112(5):677-86.

Cavill I, Kraaijenhagen R, Pradella R, D Onofrios G, Herkner K, Rowan RM, et al. In vitro stability of reticulocyte count. Clin Lab Haemat. 1996;18:9-11.

Brugnara C, Jo Hipp M, Irving PJ, Lathrop H, Lee PA, Minchello EM, et al. Automated reticulocyte counting and measurement of reticulocyte cellular indices: evaluation of the Miles H* 3 blood analyzer. Am J Clinil Pat. 1994 Nov 1;102(5):623-32.

Tichelli A, Gratwohl A, Driessen A, Mathys S, Pfefferkorn E, Regenass A, et al. Evaluation of the Sysmex R-1000®: An automated reticulocyte analyzer. Am J Clin Path. 1990 Jan 1;93(1):70-8.