Comparison of reticulocyte count by VCS technology; Beckman Coulter Hematology LH785 & fluorescent flow cytometry; XT4000i automate hematology analyzers

Kridsada Sirisabhabhorn1 Anurak Choeymang2 Supaporn Pumpa1 Naiyana poottasane1 and Palakorn Puttaruk1

1Department of Medical Technology Laboratory, Thammasat University hospital, Pathumthani, Thailand 2Pharmacology and Toxicology Unit, Faculty of Allied Health Sciences, Thammasat University, Pathumthani, Thailand

Reticulocyte count is significant for indication a new erythrocyte synthesis. Especially, usually applied for diagnosis and monitor in new born blood and anemia diseases. The traditional method for investigation of reticulocyte count with 1% brilliant cresyl blue stain have been long time practice, it not suitable for work load unit and rapid diagnosis requirement today. The new principle of reticulocyte detection was applied with 2 automate hematology analyzers which Coulter®LH785; VCS (volume, conductivity and light scatter) technology and Sysmex®XT-4000i; fluorescent flow cytometry for improve reticulocyte count diagnosis. This study propose for evaluate VCS technology by two automate hematology analyzers, compared with traditional method. All 105 cases were investigated by comparison among Coulter®LH785, Sysmex® XT-4000i and traditional method approved percentage reticulocyte count by 3 professional medical technologists. Pearson correlation was used for consider correlation. The results demonstrated that Coulter®LH785 and Sysmex® XT-4000i correlation coefficients (r) as 0.889, p-values < 0.05 and 0.936, p-values < 0.05 respectively. Moreover, the comparisons between Coulter®LH785 and Sysmex® XT-4000i illustrated correlation coefficients (r) as 0.902. The traditional method of human reticulocyte countable was not different significantly statistic analysis by ANOVAs test. Conclusion, 3 methods of Coulter®LH785, Sysmex® XT-4000i automate hematology analyzers and traditional reticulocyte counts are good correlation within p-values < 0.05. Moreover, it is helpful for work flow management than traditional method and reduce turn around time (TAT) of reticulocyte investigation.

Abstract

Key words: reticulocyte, fluorescent flow cytometry, VCS technology

Manual LH785 XT4000i

Manual Pearson Correlation 1 .889(**) .936(**)

Sig. (2-tailed) .000 .000

N 105 105 105

LH785 Pearson Correlation .889(**) 1 .902(**)

Sig. (2-tailed) .000 .000

N 105 105 105

XT4000i Pearson Correlation .936(**) .902(**) 1

Sig. (2-tailed) .000 .000

N 105 105 105

Methods & Correlations

** Correlation is significant at the 0.01 level (2-tailed).

0

1

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9

0 2 4 6 8 10

XT

40

0i

Manual 0

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0 2 4 6 8 10

LH

78

5

Manual 0

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0 2 4 6 8 10

XT

40

0i

LH785

Introduction

Materials and Methods

Complication of conventional method used for investigation % reticulocyte increase job workload and leads to long TAT (Turn Around Time) and costumer dissatisfaction. Moreover manual method needs microscope specialist for % reticulocyte counting. Today the principle of fluorescent flow cytometry, Sysmex®XT-4000i and VCS technology, Coulter®LH785 can reduce work flow of conventional method for % reticulocyte count analysis. However measuring accuracy of each method is significant for reliable report. The objective in this study was to evaluated comparative both two principle of automate analyzer above with conventional or manual method , 1% brilliant cresyl blue stain using Pearson Correlation. The study was shown benefit that confidence reliable % reticulocyte count values and provided short procedure of analysis which suitable each analytical department.

Conventional method with 1% brilliant cresyl blue stain

Fluorescent flow cytometry, Sysmex®XT-4000i

VCS technology, Coulter®LH785

Results

Conclusion and discussion

In this study conclude that two automate analyzers which composed principle of fluorescent flow cytometry, Sysmex®XT-4000i and VCS technology, Coulter®LH785 were presented good correlation for investigated % reticulocyte counting by correlation coefficients (r) values as 0.936 and 0.889 within p-values < 0.05 when compared with conventional method. Additionally two automate analyzers were demonstrated correlation coefficients (r) as 0.902 within p-values < 0.05 thus also good correlation each other. The performance ensuring in conventional method by three special medical technologists were shown no different significantly statistic analysis by ANOVAs test. The automated analyzers can provide helpful in % reticulocyte investigation and improve diagnosis approach to standard, reduced individual human errors and eliminated process of crysel blue staining and slide preparation. Lastly, quality assurance necessary should monitoring update always by medical technologist. This study declared no conflict of interest.

References

R=0.902 R=0.889 R=0.936

Conventional method

Sysmex®XT-4000i Coulter®LH785

Graph 1. Illustrated correlation curve of % reticulocyte count between Sysmex®XT-4000i and conventional method (manual method)

Graph 2. Illustrated correlation curve of % reticulocyte count between Coulter®LH785 and conventional method (manual method)

Graph 3. Illustrated correlation curve of % reticulocyte count between Coulter®LH785 and Sysmex®XT-4000i

Pearson Correlations

Table 1. Illustrated correlation results 3 method for investigated percent of reticulocyte count which compared using Pearson

correlation, p-values < 0.05

All 105 cases were investigated % reticulocyte count with 3 principles mention above. All methods were investigated their relationships using Pearson correlation, which explored correlation coefficients (r) p-values < 0.05. Using 3 medical technologists who were ensued not different significantly statistic analysis by ANOVAs test in conventional method evaluated mean values of % reticulocyte count

Sysmex. Reticulocytes and their significance. Sysmex Xtra Online, 2010: 1-12. PCL Holding Co., Ltd. Standard Operating Procedure ( SOP ) For Coulter ReticPakTM