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ISLH

Laboratory Hematology 7:125129


2001 Carden Jennings Publishing Co., Ltd.

Official Publication

An Evaluation of the Beckman Coulter Compact


5-Part Differential Hematology Analyzer AcT 5diff
CAROLINE KERN
Haematology Laboratory, Appensell Kantonal Hospital, Herisau, Switzerland

a 5-part differential leukocyte count (DLC) and to generate a


panel of comprehensive flags in cases of abnormal results. During 1 month we evaluated the AcT 5diff analyzers CBC in
comparison with the 3-part differential instrument currently
used in our laboratory, the CD 1600 (Abbott Laboratories,
Abbott Park, IL). We also evaluated the AcT 5diff DLC in
comparison with the reference manual method and performed
time stability studies on CBC and DLC parameters.

ABSTRACT

The Beckman Coulter compact 5-part differential hematology analyzer, the AcT 5diff analyzer, was evaluated. It
provides up to 26 parameters with a throughput of 60 samples/hour on a selectable mode: complete blood count
(CBC) or CBC plus 5-part differential leukocyte count
(DLC), with a minimal sampling size (53 L for CBC plus
DLC and 30 L for CBC). The AcT 5diff analyzer was
evaluated on the basis of a simplified protocol based on the
routine workload. The sample stability was excellent for all
CBC parameters (up to 72 hours) and for DLC parameters
(up to 48 hours) with a slight shift on monocyte counts.
Correlation coefficients for all CBC parameters were excellent for neutrophils, lymphocytes, and eosinophils and
acceptable for monocytes. The clinical sensitivity of the
white blood cell flagging system performed well with a sensitivity of 92.8% and a specificity of 87.2%. The instrument was easy to use and offered a sample-by-sample selection for CBC only or CBC plus DLC. Lab. Hematol.

MATERIAL AND METHODS


System Description
The AcT 5diff analyzer is a small bench-top hematology
system that can analyze up to 60 samples per hour and uses a
30-L sample in CBC and a 53-L sample in CBC/DLC
mode. It counts white blood cells (WBC), red blood cells
(RBC) and platelets (PLT) by the impedance method.
Hematocrit (Hct) is measured by the sum of RBC counted
in a specified volume of diluted blood. The leukocyte differential count is performed in 2 different channels:

Flow analysis by means of 2 measurements realized


on each cell independently: the volume by impedance method and light scatter after contact of WBC
and Chlorazol Black E stain (Beckman Coulters
AcV technology) [1].

A basophil channel in which the total leukocyte


count is performed after mixing with a specific
reagent. This reagent gently strips the leukocyte
membranes and preserves the basophil membranes.

2000;7:125129

KEY WORDS:

5-Part differential Hematology


analyzer AcT 5diff

INTRODUCTION
The AcT 5diff analyzer (Beckman Coulter, Miami, FL) is a
highly compact automated hematology analyzer designed to
provide the parameters of a complete blood count (CBC) and

Blood Samples
Samples drawn into 3-mL K3 EDTA vacuum tubes were
selected on a random basis from our routine workload over a
4-week period. All samples were analyzed on both instruments within 4 hours of draw. Five normal samples were
kept at room temperature for the stability study.

Received January 5, 2001; accepted April 23, 2001


Correspondence and reprints: Caroline Kern, Kantonales Spital, Central
Laboratory, 9100 Herisau, Switzerland

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126

C. Kern

Long-term Sample Stability


Five patient samples were analyzed on the AcT 5diff analyzer at selected intervals over a 72-hour period. An initial
baseline was established at 1 hour after draw. The samples
were stored at room temperature and analyzed after 9, 24,
32, 48, and 72 hours. Variations for each parameter are
expressed as a ratio:
Mean Baseline Mean Time hours / Mean Baseline.

CBC and Leukocyte Differential Comparison


Seventy-five (normal and abnormal) samples were tested
on the AcT 5diff analyzer and on the CD 1600. The CD
1600 was considered the reference instrument for the CBC
parameters. It is the laboratorys routine instrument, and we
tested its performance during a pre-acquisition internal evaluation. On every sample a blood film was spread and stained
using May Grunwald Giemsa. A 100-cell manual differential
was performed on each slide by an experienced technologist.
The AcT 5diff analyzer differentials were correlated against
the manual differentials. The CBC and differential parameters were compared by regression analysis.
Clinical Sensitivity
Manual and AcT 5diff analyzer differentials were compared to determine the clinical sensitivity of the instrument.
Each sample was classified in the following categories:
True negative (TN) Negative on the blood film, no flag
on the instrument.
True positive (TP)
Positive on the blood film, presence of flags on the instrument
screen and on report.
False negative (FN) Positive on the blood film, no flag
on the instrument.
False positive (FP)
Negative on blood film, presence
of flags on the instrument.

The following ratios were calculated:


Sensitivity TP/(TP + TN)
Specificity TN/(TN + FP)
Efficiency (TP + TN)/Total

RESULTS
Long-term Stability
CBC parameters were very stable over the 72-hour period (Table 1). The differential parameters showed acceptable stability. Neutrophil and lymphocyte values were stable up to 48 hours. A minor shift occurred in neutrophil
and lymphocyte populations 9 hours after sampling with
no clinical significance. The monocytes increased slightly
after 24 hours.
CBC Correlation
As shown in Figure 1, there was excellent CBC correlation, with all correlation coefficients greater than 0.98 except
mean corpuscular volume (MCV), which exhibited an
acceptable correlation, r = 0.93.
Leukocyte Differential Count Comparison
All leukocyte differential parameters of the AcT 5diff
analyzer showed acceptable results when compared to the
100-cell manual differential (Figure 2). Results were in
accordance with those reported by other authors [2,3].
Monocytes showed a lower correlation coefficient without
clinical significance. Other authors have reported such differences between monocyte counts on automated instruments versus manual counts [4].
Clinical Sensitivity
During this evaluation, positive samples were identified
on the AcT 5diff analyzer when any of the leukocyte flags
were present. Samples were identified as positive by the

TABLE 1. Long-term Stability


Ratio*
Parameter
White blood cells, 109/L
Red blood cells, 1012/L
Hemaglobin, g/L
Hematocrit, L/L
Mean corpuscular volume, fL
Red cell distribution width, %
Platelets, 109/L
Neutrophils, 109/L
Lymphocytes, 109/L
Monocytes, 109/L
Eosinophils, 109/L

Mean at baseline

9 hours

24 hours

32 hours

48 hours

72 hours

7.57
4.12
114
0.345
84.2
13.7
369
4.62
1.97
0.61
0.23

0.03
0.01
0.00
0.02
0.01
0.02
0.02
0.11
0.13
0.03
0.03

0.03
0.00
0.01
0.00
0.00
0.01
0.04
0.03
0.03
0.03
0.03

0.03
0.01
0.00
0.01
0.01
0.01
0.01
0.05
0.01
0.16
0.38

0.02
0.00
0.01
0.01
0.01
0.01
0.02
0.01
0.10
0.15
0.02

0.01
0.00
0.01
0.01
0.01
0.01
0.06
0.22
0.22
0.48
0.01

*Mean Baseline Mean Time x hours / Mean Baseline.


Of 5 samples.

Evaluation of the Beckman Coulter AcT 5diff

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FIGURE 1. Regression analysis for complete blood count parameters in a comparison of the AcT 5diff and the CD 1600. A, Red
blood cells (RBC). B, White blood cells (WBC). C, Platelets (PLT). D, hemoglobin. E, Mean corpulscular volume (MCV).

manual differential when one or more of the criteria of positivity (Table 2) were met.
Table 3 shows the clinical sensitivity of the AcT 5diff and
the CD1600 analyzers in this study.

DISCUSSION
The AcT 5diff hematology analyzer provided complete
CBC and leukocyte differential using impedance (Coulter
Principle) and cytochemistry leading to specific staining of
cell granules.
All CBC parameters were stable up to 72 hours and differential parameters were stable up to 48 hours. This result is
of high value for centers that receive samples from outside
collection points because such samples usually travel several
hours prior to being tested.
Prior to the comparison between the 2 analyzers, each
instrument was calibrated using calibrator bloods according
to manufacturer recommendations. Excellent correlation was

found between instruments for all CBC parameters. Slight


biases have been noticed due to differences in the technologies applied to the automatic counts in the 2 instruments.
Good correlation for DLC parameters, compared to the
manual method, has been demonstrated. Minor differences
between the manual and the instrument DLC were found.
Those differences are without clinical significance and have
been previously reported in the literature [5].
The evaluation of the WBC flagging system indicated
high sensitivity (92.8%). The only 2 false negative results
observed in this study were due to the presence of a moderate number of band cells for one sample and a few reactive
lymphocytes in the other. Specificity was good (87.2%), with
an overall efficiency of 89.3%.
Working on the CD1600, we reviewed almost every sample, knowing that this 3-part differential analyzer usually
failed to detect low to moderate numbers of reactive lymphocytes and immature granulocytes. During the present study,
the AcT 5diff analyzer, by its sensitivity to detect abnormal

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C. Kern

FIGURE 2. Regression analysis for differential leukocyte count in a comparison of the AcT 5diff and manual DLC. A, Neutrophils.
B, Lymphocytes. C, Monocytes. D, Eosinophils.

cells, led to an increase in the global productivity of the


hematology division by drastically reducing the number of
unnecessary smears.
The low sample volume the instrument used for CBC
and DLC (53 L) and very low volume (30 L) for CBC
was of real advantage when testing pediatric blood samples.
The system was very easy to use and offered a choice
between CBC-only mode, in which the reagents for DLC
were saved, and CBC-plus-DLC mode.

The AcT 5diff analyzer required no routine daily maintenance. A daily automatic shutdown ensured the proper
cleaning of the system. Overall, this instrument was perceived as very convenient for a laboratory performing about
40 to 50 samples a day.
The AcT 5diff analyzer model we tested was an open-vial
system; a model with closed-vial sampling and a database
management system is also available.
TABLE 3. Morphological Classification
AcT 5diff

CD 1600

75
26
2
41
6
92.8%
87.2%
89.3%

75
8
21
42
4
27.6%
91.3%
66.6%

TABLE 2. Criteria of Positivity on a Blood Smear Examination


Type of cell

Blasts
Promyelocytes
Myelocytes
Reactive lymphocytes
Erythroblasts

1
1
>2
2
2

Total
True positive
False negative
True negative
False positive
Sensitivity
Specificity
Efficiency

Evaluation of the Beckman Coulter AcT 5diff

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3. Butarello M, Gadott M, Lorenz C. Evaluation of four automated

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hematology analyzersa comparative study of differential counts.


Am J Clin Pathol. 1992;97:345-352.
4. Goossens W, Van Hove L, Verwilghen Rl. Monocyte counting
discrepancies in results obtained with different automated instruments. J Clin Pathol. 1991;44:224-227.
5 Dacie JV, Lewis SM. Differential Leukocyte Count: Practical Hematology, 7th ed. New York: Churchill Livingstone; 1991.