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Vet Clin Pathol. Author manuscript; available in PMC 2009 July 17.
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Vet Clin Pathol. 2008 December ; 37(4): 373–384. doi:10.1111/j.1939-165X.2008.00085.x.
Comparative clinical study of canine and feline total blood cell

count results with seven in-clinic and two commercial laboratory
hematology analyzers
Martina Becker1, Andreas Moritz1, and Urs Giger2

1Department of Veterinary Clinical Sciences, Clinical Pathology, and Clinical Pathophysiology,
Justus-Liebig University Giessen, Giessen, Germany

2Section of Medical Genetics, School of Veterinary Medicine, University of Pennsylvania,
Philadelphia, PA, USA
Abstract
Background—A CBC is an integral part of the assessment of health and disease in companion
animals. While in the past newer technologies for CBC analysis were limited to large clinical
pathology laboratories, several smaller and affordable automated hematology analyzers have been
developed for in-clinic use.
Objectives—The purpose of this study was to compare CBC results generated by 7 in-clinic laser-
and impedance-based hematology instruments and 2 commercial laboratory analyzers.
Methods—Over a 3-month period, fresh EDTA-anticoagulated blood samples from healthy and
diseased dogs (n = 260) and cats (n = 110) were analyzed on the LaserCyte, ForCyte, MS45, Heska
CBC, Scil Vet ABC, VetScan HMT, QBC Vet Autoread, CELL-DYN 3500, and ADVIA 120
analyzers. Results were compared by regression correlation (linear, Deming, Passing-Bablok) and
Bland-Altman bias plots using the ADVIA as the criterion standard for all analytes except HCT,
which was compared with manual PCV Precision, linearity, and carryover also were evaluated.
Results—For most analytes, the in-clinic analyzers and the CELL-DYN performed similarly and
correlated well with the ADVIA. The biases ranged from −0.6 to 2.4 × 109/L for WBC count, 0 to
0.9 × 1012/L for RBC count, − 1.5 to 0.7g/dL for hemoglobin concentration, −4.3 to 8.3 fL for MCV,
and −69.3 to 77.2 × 109/L for platelet count. Compared with PCV, the HCT on most analyzers had
a bias from 0.1% to 7.2%. Canine reticulocyte counts on the LaserCyte and ForCyte correlated but
had a negative bias compared with those on the ADVIA. Precision, linearity, and carryover results
were excellent for most analyzers.
Conclusions—Total WBC and RBC counts were acceptable on all in-clinic hematology
instruments studied, with limitations for some RBC parameters and platelet counts. Together with
evaluation of a blood film, these in-clinic instruments can provide useful information on canine and
feline patients in veterinary practices.
Keywords
Cat; dog; hematology; laboratory method; platelet; red blood cell; white blood cell
© 2008 American Society for Veterinary Clinical Pathology

Correspondence Dr. Martina Becker, Department of Veterinary Clinical Sciences, Clinical Pathology and Clinical Pathophysiology,
Justus-Liebig University Giessen, Frankfurter St. 126, 35392 Giessen, Germany E-mail: E-mail: martina.becker@vetmed.uni-giessen.de.
Becker et al. Page 2
Introduction
The evaluation of blood cells is an integral part of the routine assessment of healthy and
diseased companion animals. A PCV and the microscopic examination of a blood film still
form the cornerstones of clinical veterinary hematology, but quantitative counts of different
blood cells are also needed to better assess an animal’s health status. Manual counting of blood
cells by microscopic counting chambers is too tedious and inaccurate for routine practice. The
first automated impedance blood cell counter was introduced by Coulter half a century ago.1
Since then, hematology instruments have evolved to use laser flow cytometry and cytochemical
methods for blood cell quantitation. While in the past these newer technologies were limited
to large clinical pathology laboratories, several smaller and affordable automated hematology
analyzers have been developed for in-clinic use.
The parameters measured by different hematology analyzers vary, and the practical usefulness
and accuracy of the test results of different in-clinic and larger commercial clinical pathology
laboratory (subsequently referred to as laboratory) instruments have not been comprehensively
assessed and compared among each other. One inherent difficulty is that there is no single gold
standard or true automated reference method as each instrument has advantages and limitations.
The QBC VetAutoread was one of the first instruments introduced in practice and is still a
commonly used in-clinic hematology analyzer, although clinical studies revealed several
limitations compared with commercial laboratory methods.2–7 Only a few studies have been
carried out to compare other in-clinic hematology analyzers to reference laboratory methods.
8–16 In addition, no comparisons on the flags and their accuracy and clinical importance have
been published in veterinary medicine.
The purpose of this clinical study was to compare the results of CBCs from dogs and cats on
7 in-house and 2 laboratory hematology analyzers. We also considered the ease of clinical use,
accuracy, flags, advantages, and limitations of the various hematology systems.
Materials and Methods

From December 2004 to February 2005, 2-mL EDTA-anticoagulated blood samples were
collected for routine in-house blood testing from healthy (spay or castration presurgical
examination, >6 months old) and diseased adult dogs and cats presented to 3 small animal
clinics in the Fort Lauderdale area (Florida, USA). After making 2 freshly prepared blood films,
the blood samples were stored at room temperature (22°C) for up to 3 hours until being
transported by courier to a nearby laboratory (Clinical Diagnostic Solutions Inc., Plantation,
FL, USA). All in-clinic analyzers (see Table 1) and the CELL-DYN 3500 were available for
immediate analysis in this laboratory, while an aliquot of each blood sample was sent overnight
on ice to another laboratory (IDEXX Laboratories Inc., Westbrook, ME, USA) for analysis on
the ADVIA 120 analyzer (Siemens, Deerfield, IL, USA).
All in-clinic hematology analyzers to be assessed in this study were set up with a current
software version for small animals in an air-conditioned laboratory. Technical details for each
hematology instrument are summarized in Table 1. Maintenance procedures on all analyzers
were performed according to the manufacturers’ instructions and, when available, appropriate
control samples were run daily. In addition, a manual PCV was determined at the Clinical
Diagnostic Solutions laboratory using microhematocrit centrifugation (StatSpin VT
Centrifuge, Statspin Inc., Norwood, MA, USA).
All samples were analyzed within 6 hours after blood collection, except for analysis on the
ADVIA, which was performed within 24 hours. Before analysis, tubes were placed on a roller
plate (Coulter Mixer, Coulter Electronics Ltd., Fullerton, CA, USA) for at least 1 minute to
ensure proper mixing. All blood samples were sequentially analyzed by 1 of the authors (M.B.)
in a specific order on all of the instruments (excluding the ADVIA), except when there was
insufficient sample volume, for repeated analyses, or when one of the instruments was
malfunctioning. In all samples for which 1 or more of the instruments reported
thrombocytopenia, an estimation of the platelet (PLT) count on the blood smear was performed
by 1 of the authors (M.B.). If PLTs were clumped, “clumped platelets” were reported. If PLT
were evenly distributed on the blood film, the mean number of PLT per field in 10 × 100
objective fields was multiplied by 20,000 to estimate PLT numbers/μL.17 When PLT counts
were ≤ 50 × 109/L, the thrombocytopenia was classified as severe.
Precision, linearity, and carryover studies were performed on all analyzers except the QBC
and the ADVIA. A 700-mL sample of K-EDTA-anticoagulated blood from 2 healthy dogs was
collected and pooled (blood was supplied by a commercial blood bank). Between analyses the
blood samples were stored at room temperature. A 2-mL aliquot of well-mixed pooled blood
was prepared and run on each instrument. To eliminate the effects of cell aging, the next run
was started when all instrument cycles were finished. The remaining volume of pooled blood
was mixed well, divided into 50-mL aliquots, and centrifuged at room temperature at 100g for
20 minutes. After centrifugation, plasma was removed and pooled into 1 tube, which was
labeled as level 1. The buffy coats also were removed and pooled into 1 tube, which was labeled
as level 5. For the linearity study, 3 additive levels were prepared by diluting level 5 with level
1 in ratios of 1:3 (level 2), 1:1 (level 3), and 3:1 (level 4). For each instrument, 5 different tubes
containing 2 mL of each level were prepared. Each level was analyzed 2 times followed by 1
analysis of level 5. The order of analyses of different levels was randomized. For carry-over
studies, tubes were filled with 2 mL of level 5 and analyzed twice by each instrument followed
by 3 analyses of phosphate-buffered saline (PBS; separate tube of PBS for each analysis). This
sequence was repeated 3 times. As the VetScan is not able to measure PBS, the previously
prepared level 2 was used instead.
Statistical analysis
Data were collected electronically from each hematology instrument and directly transferred
to a Microsoft Excel spreadsheet (Microsoft Corp., Redmond, WA, USA) for analysis. PCV
data and data generated by the ForCyte and QBC instruments were entered manually. Results
were statistically analyzed by linear regression, Deming regression (normally distributed
measurement errors), Passing-Bablok regression (nonnormally distributed measurement
errors) and Bland–Altman plots using Microsoft Excel with Analyse-it (Analyse-it Software
Ltd., Leeds, UK) and GraphPadPrism version 4.0 (GraphPad Software Inc., La Jolla, CA,
USA).18,19 Results for MCH, MCHC, red cell distribution width, and nucleated RBCs were
not evaluated in this study.
Correlation coefficients (r) and the SD of the residuals (Sy|x) from linear regression analyses,
the intercept and slope with 95% confidence intervals calculated by Deming regression or
Passing-Bablok regression, and biases with 95% limits of agreement calculated by Bland–
Altman plots were reported for each instrument and analyte. Bland–Altman plots and the
calculated intercept and slope were used to characterize the source of bias. When r is < .975,
the estimate of intercept and slope may not be accurate to determine the source of error20;
hence the Bland–Altman plots were solely used. To assess the amount of random error, the SD
obtained in the replication experiment, Sy|x, and 95% limits of agreement from the Bland–
Altman analysis were evaluated.
Accuracy was determined by comparing results to those of the ADVIA, which was defined as
the reference method in this study. Accuracy was assessed separately for canine and feline
WBC, RBC, PLT, and reticulocyte counts as well as for MCV and hemoglobin (HGB)
concentration for each in-clinic instrument and the CELL-DYN. The calculated HCT on all
instruments was compared to the PCV as the reference method.
The SD and coefficients of variation (CVs) were calculated for the replication experiments.
With respect to the linearity study, values for levels 2, 3, and 4 were determined for each
instrument based on the contributions of levels 1 and 5 (as determined by the CELL-DYN) to
each of the other levels. Thereafter, the mean of the measured values was compared to the
calculated result using regression analysis. Carry-over was calculated by subtracting the mean
of the third run of PBS from the mean of the first run of PBS and dividing by the mean of level
5 for WBC, RBC, PLT, and HGB.
Results
The handling of all in-clinic instruments was generally simple and user friendly. The
instruments functioned well throughout the study and required maintenance procedures were
minimal. The Heska CBC and MS45 became blocked several times throughout the study and
internal tubes and filters had to be changed. The VetScan had to be replaced once during the
study when a problem with the power supply occurred. The LaserCyte had a significantly
longer cycle time per sample than the other instruments and the QBC had a longer sample
preparation time before analysis.
Linearity, precision, and carry-over

The linearity of all instruments was excellent (r > .993) over a broad range of HGB
concentrations (0–22 g/dL) and WBC (0–80 × 103/µL), RBC (0–8.5 × 106/µL), and PLT (0–
1500 × 103/µL) counts, except for WBC count on the Scil Vet ABC (r = .970), which displayed
a flag for counts > 40 × 103/µL to indicate that the replicate counts exceeded the system’s
precision limit. The CVs of the in-clinic analyzers ranged from 0.7% to 4.1% for WBC, RBC,
HGB, and HCT (Table 2). In contrast, the CV for MCV was ≤ 1.0%, and for PLT ranged from
4.4% to 9.2%. CVs of the CELL-DYN ranged from 0.5% to 2.2%, depending on the parameter.
Carry-over values of < 0.25% were achieved with all instruments for all cell counts and HGB
concentration, except for the VetScan (RBC 3.6%, HGB 0.8%, PLT 14.1%) and the CELL-
DYN (PLT 0.4%).
Flags
The percentage of flagged samples varied among the different instruments (Table 3). The Heska
analyzer flagged PLT count in 89% of feline samples because of inappropriate size separation
between RBCs and PLTs; in 36% of all measurements the instrument recommended reanalysis.
The same reason was responsible for the 44% of PLT counts marked with a flag by the Scil
Vet ABC (however, whereas the Heska uses a floating threshold to separate RBC and PLT,
the Scil Vet ABC applies a fixed threshold). Error messages on the QBC were mostly due to
PLT clumps or other cells on top of the float. The ForCyte displays 2 error messages: “rerun”
and “slide”; 20% of feline samples were flagged with the error message “slide.” We concluded
that slide review was being recommended in these cases; however, the manufacturer of the
instrument did not offer any further information concerning a possible cause. The CELL-DYN
flagged a high percentage of WBC counts due to differences in results between the WBC
impedance channel (WIC) and WBC optical channel (WOC). Depending on the parameter,
33% to 80% of samples analyzed on the ADVIA were marked with an error message.
To further assess the flags, we evaluated how many samples were marked with an error message
by > 1 instrument. Two instruments flagged the WBC count in 30% of canine samples and ≥3
instruments flagged WBC count in 10% of samples. Two instruments flagged PLT count in
27% of canine samples, and ≥2 instruments flagged PLT count in 3% of samples. Two
instruments flagged WBC count in 28% of feline samples and >2 instruments flagged WBC
count in 25% of samples. Two instruments flagged PLT count in 37% of feline samples and
≥3 instruments flagged PLT count in 31% of samples.
To verify the value of the flags, we selected parameters and instruments with a high percentage
of flagged samples for further analysis, and separated the flagged and unflagged results (Figure
1). Interestingly, a number of flagged values appeared to correlate well between in-clinic and
laboratory methods and thus were likely accurate, while values of some non-flagged samples
differed between in-clinic and laboratory results.
Accuracy
RBC parameters—PCV ranged from 8% to 72% (mean 41%, median 43%) in canine
samples and from 5% to 49% (mean 32%, median 33%) in feline samples. Seventy-three canine
samples had a PCV< 37% and 22 feline samples had a PCV< 24%; 17 canine samples had a
PCV ≥ 55% and 6 feline samples had a PCV > 45%. Correlations between all in-clinic
instruments and the ADVIA were excellent for all RBC parameters except MCV in dogs and
several parameters in cats (Table 4).
All in-clinic instruments (except the QBC) and the CELL-DYN had a positive bias for HCT
in comparison with manual PCV (Table 5). The positive biases were due mostly to constant
systematic errors (Supporting information File 1, Figures S.1.1–S.1.36). An extremely high
bias was observed for HCT on the Vet-Scan with samples from both species and was associated
with a strong positive bias for MCV (due to constant systematic error) compared with the
ADVIA (Supporting information Files 1 and 2, Figures S1.21–S.1.24 and S2.17–S2.20). A
similar association was found for the CELL-DYN with feline samples. The Sy|x− and the 95%
limits of agreement were relatively high for all instruments, except the QBC and ADVIA.
For RBC counts, the ForCyte and Heska had a moderate positive bias for both species, whereas
the MS45, Scil Vet ABC, and CELL-DYN had moderate positive bias only for feline samples.
The positive biases were due mostly to combined systematic errors (mixture of constant and
positive proportional errors) (Supporting information File 3, Figures S3.4, S3.11–S3.16,
S3.21–S3.24, S3.27, and S3.28). The Sy|x− and 95% limits of agreement of the Bland-Altman
analysis were similar for the different instruments. The biases for the LaserCyte and VetScan
were close to zero. Biases for HGB were small, except with the MS45 and QBC, which had a
negative bias due to constant systematic error (Supporting information File 4, Figures S4.9–
S4.12 and S4.29–S4.32). The Sy|x− and 95% limits of agreement were relatively narrow with
all instruments.
WBC count—Based on results from the ADVIA, WBC counts ranged from 0.98 to 67.00 ×
109/L (mean 16.21 × 109/L, median 11.44 × 109/L) in canine samples and 1.73 to 55.89 ×
109/L (mean 13.81 × 109/L, median 11.97×109/L) in feline samples. Fourteen samples from
dogs had WBC counts < 6000 × 109/L and 10 samples from cats had WBC counts < 5500 ×
109/L. Sixty-one samples from dogs had WBC counts > 17,000 × 109/L and 17 samples from
cats had WBC counts > 15,000 × 109/L. Correlations for WBC counts were excellent among
all instruments for both species (Table 4). Biases were close to zero, except for the QBC, which
had slightly higher biases for canine samples due to a combined systematic error, and for feline
samples due primarily to a positive proportional error (Supporting information File 5, Figures
S5.30, S5.32).
PLT count—Based on results from the ADVIA, PLT counts ranged from 46 to 725 × 109/L
(mean 301 × 109/L, median 285 × 109/L) in canine samples and from 16 to 858 × 109/L (mean
289 × 109/L, median 266 × 109/L) for feline samples. In 47 canine and 23 feline samples, PLT
counts were < 200 × 109/L; in 18 canine samples PLT count was > 500 × 109/L and in 4 feline
samples PLT count was > 600 × 109/L. Correlation coefficients for PLT counts were fair to
good (r = .685−.890) for all in-clinic instruments and the CELL-DYN in both species. The Scil
Vet ABC had a strong negative bias for PLT count in both species, due mostly to constant
systematic errors (Supporting information File 6, Figures S6.26 and S6.28). The VetScan had
a strong negative bias for PLT count in feline samples due to a strong negative proportional
error (Supporting information File 6, Figure S6.20). The Sy|x− and 95% limits of agreement
were unfavorable for all in-clinic instruments and the CELL-DYN. The number of samples
with severe thrombocytopenia was low (n = 14; 10 canine and 4 feline samples); all in-clinic
and laboratory instruments correctly identified severe thrombocytopenia in a maximum of 70%
(7/10) of canine and 75% (3/4) of feline samples.
Reticulocyte count—Two in-house instruments (LaserCyte and ForCyte) and the ADVIA
were able to determine absolute reticulocyte counts in canine samples (reagents for the CELL-
DYN were not available at the time of the study). In canine samples, reticulocyte counts ranged
from 0 to 647 × 109/L (126 samples had ≥60 × 109/L), which correlated with the amount of
polychromasia. Correlation coefficients were fair for both the LaserCyte (r = .789) and ForCyte
(r = .662). Sy|x-values were between 25 and 30 × 109/L for both instruments, indicating
moderate random error. Both analyzers had a negative bias for reticulocyte counts (LaserCyte
−36 × 109/L, ForCyte −62 × 109/L), compared with the ADVIA, due mostly to a proportional
systematic error, and preventing determination of 95% limits of agreement. Feline reticulocyte
counts were measured only on the ADVIA and the LaserCyte (ForCyte software for feline
reticulocyte counts was not available at the time of data collection). In feline samples, good
correlation of reticulocyte counts was found between the LaserCyte and ADVIA (r=.862). The
Sy|x− was 20 × 109/L, with 95% limits of agreement of −49 to 48 × 109/L and negligible bias
(−0.75 × 109/L).
Discussion
Practitioners have the option of performing certain laboratory diagnostic tests in-clinic or
submitting samples to a commercial clinical pathology laboratory. While PCV and blood film
analyses provide crucial information, a CBC is often deemed necessary for the assessment of
health and disease of companion animals. The selection of an automated hematology analyzer
for a particular clinic depends on many factors, including the urgency of having immediate
access to test results for a patient, number of samples to be analyzed per day, desired analytes
measured by an instrument, ease of use (vs shipping samples to a laboratory), purchase price
and running expenses, and accuracy of the results.21
The accurate and precise measurement of blood cells and associated parameters is challenging
because of the complex and variable nature of blood cells and blood cell morphology. Despite
known limitations of large commercial analyzers such as the CELL-DYN and the ADVIA,
their accuracy is considered to be high because of the large number of cells analyzed and
extensive validation studies. Furthermore, trained laboratory technicians usually operate these
instruments, quality assurance programs are in place, and blood films are generally examined
to confirm (at least abnormal) results in laboratories where these analyzers are used. Finally,
clinical pathologists often are at hand to review test results and blood films. Commercial
laboratory services, however, may not be available in all geographic locations and delays may
occur in getting test results.
The International Council for Standardization in Hematology (ICSH) defines accuracy as “a

measure of agreement between the estimate of value and the true value” using independent
techniques.22 The only analytes that can be measured accurately are HGB concentration, PCV,
RBC count, total WBC count, and differential WBC counts. Alternatively, comparability
between analyzers can be assessed. This is defined as the ability of the instrument to produce
results that agree satisfactorily with those obtained by procedures in routine use.22
Comparability is an accepted method to evaluate hematology instruments; however, in the case
of differences between methods, it is necessary to identify which method is inaccurate. For
practical purposes, we chose the ADVIA, a dual optical- and peroxidase-based hematology
analyzer as the reference instrument for our study. This instrument is widely used and accepted
in veterinary clinical pathology laboratories and has been validated for animal samples. We
also included the CELL-DYN in this study, an impedance- and laser-based hematology
analyzer that is also widely used and validated in veterinary medicine. The results for the in-
clinic instruments were similar when they were compared with either laboratory instrument
(data not shown); however, the ADVIA was used as the criterion standard. Interestingly, the
CELL-DYN was not more accurate than the in-clinic instruments when compared with the
ADVIA, reflecting the overall limitations of any hematology analyzer, whether for in-clinic
or laboratory use.
The delay of up to 24 hours in the analysis of samples analyzed on the ADVIA was a limitation
of the present study; however, such a delay is typical in a clinical setting and previous studies
have shown that valid reproducible results are obtained with the ADVIA on samples up to 24
hours old from healthy dogs and cats.23 After 24 hours of storage at 4°C, a maximal mean
deviation of 3% was found for RBC and WBC parameters and accuracy was considered
sufficient until 36 hours after collection. However, for PLTs, the deviation in results after 24
hours reached 20%. The ADVIA also produces many flags with stored samples,24 which was
observed in this study.
Similar to most method comparison studies, we performed linear regression analyses on the
results.25 However, the correlation coefficients obtained can only be used to determine the
acceptability of the data used to calculate the regression statistics and do not ensure accuracy.
26 Therefore, Deming and Passing-Bablok regression analyses, which provide the intercept
and slope, were used; when r was low, Bland–Altman analysis also was applied. This allowed
differentiation of proportional and constant systematic errors. This was important, as
proportional errors can be reduced readily by modifying calibration factors, whereas constant
errors require the establishment of instrument-specific reference intervals. Based on the results
of the present study, analyzer-specific reference intervals are likely needed for the HCT of
some instruments (eg, the CELL-DYN, MS45, Heska CBC, VetScan, ForCyte, and Scil Vet
ABC), for HGB concentration on the MS45 and QBC, and for MCV on the VetScan. The
advantage of Deming regression over Passing-Bablok regression is the calculation of Sy|x,
which, comparable to the 95% limits of agreement calculated with Bland–Altman analysis,
gives an estimate of the random error.
Assessing the accuracy of hematology analyzers also depends on medical decision limits and
clinical requirements. Whereas several recommendations are available for human medicine
(eg, Clinical Laboratory Improvement Amendments27), no such data exist in veterinary
medicine. Thus, assessment of the accuracy of laboratory tests needs to be based on clinical
requirements. It was reassuring that total WBC counts were comparable between in-clinic and
laboratory analyzers and acceptable whether impedance or optical technology was used. The
QBC was a noteworthy exception, as it produced relatively high biases for WBC counts in
samples from both dogs and cats. This WBC bias may lead to misinterpretation and be clinically
important in patients with leukopenia. Previous studies evaluating the accuracy of the QBC
reported similar problems with the measurement of low WBC counts.2 Detailed analyses of
samples with leukopenia and neutropenia are in progress in our laboratory (M.B., unpublished
data).
All instruments except the QBC, which determines a PCV, had a positive bias for HCT.
Although the underlying cause of the positive bias remains unclear, modifications of software
settings and calibration factors could likely minimize the differences. The positive bias for
HCT values on the VetScan (5.73% in canine and 7.18% in feline samples) was comparable
to that reported for dogs in a previous study (7.9%).9 Thus, instrument-specific reference
intervals are recommended to avoid clinically relevant misinterpretations. Of more concern

were the wide 95% limits of agreement and high Sy|x for HCT on all instruments (except the
QBC), which may reflect imprecision and thus random error. Random error was likely caused
by morphologic changes in RBCs from healthy and diseased animals, because it was not
reflected in the CVs in our replication experiments (which ranged from 1.6% to 3.9% for in-
clinic instruments) when only 2 samples were used for analysis. Nevertheless, it is good and
common practice to verify the calculated HCT of a CBC with a PCV as well as to apply the
simple equation of HGB × 3 as an estimate of the HCT and PCV as long as there is no
hemoglobinemia or hypochromia.
As manual PLT counts are time-consuming, impractical, and imprecise, instrument PLT counts
are highly desirable. However, accurate determination of canine and particularly feline PLT
counts appears to be extremely challenging for in-clinic as well as laboratory analyzers due to
the overlap between RBCs and PLTs on histograms. Therefore, it may not be surprising that
all of the in-clinic analyzers and both the CELL-DYN and ADVIA missed at least 25% of
samples with severe thrombocytopenia in comparison with PLT estimates on blood smears.
The extremely high biases for PLT counts by some instruments have to be interpreted with
caution, as the blood samples were 24 hours old when measured by the ADVIA and PLT counts
can change more rapidly over time (within a few hours to a day) than other blood cell
parameters.23 Hence, the inaccuracy of the in-house instruments was likely also due to
inaccuracy of the reference method, and further investigation with other reference methods is
required. In a previous comparison study, the VetScan had low bias and excellent correlation
for PLT counts in canine samples.9 This underlines our suspicion that inaccuracy of our
reference method contributed to the unfavorable results for PLTs in our study. An advantage
of the ADVIA is its ability to detect sizable PLT clumps, thereby alerting the user of a
potentially falsely low PLT count.23 PLT estimation in blood smears is always recommended
to confirm any instrument PLT count.
Because there were high numbers of erroneously flagged samples for some parameters, and
missing flags for some questionable test results, the error messages on all of the in-clinic
instruments appear to be of limited use in clinical practice. Improvements in flagging rules and
more detailed information on how to handle flagged samples are needed for all of the
hematology instruments. Based on the results of our study, we recommend that blood smears
be evaluated briefly for all blood samples, regardless of whether a flag is displayed.
Precision reflects the ability of an assay to repeatedly obtain the same result with the same
sample. The need for analytical precision is dependent on the acceptable degree of random
variation.17 The results in our study slightly exceeded the recommendations for allowable error
for human samples,25 but were comparable to those obtained with the ADVIA in previous
studies with animal samples.23 The slightly higher imprecision of the RBC count by the
LaserCyte compared to other instruments has been reported in a previous study28 and might
be explained by the laser technology, as mild fluctuations in noise level of the laser are expected
and cannot be avoided.
In the range of cell counts tested, all of the hematology instruments had excellent linearity
except for the Scil Vet ABC at WBC counts >45 × 103/µL. All instruments had negligible
carryover of WBCs, RBCs, PLTs, and HGB, which is important to ensure that cells are not
carried into the next patient’s sample where they can falsely increase cell counts. The increased
carryover seen with the VetScan was most likely caused by imprecision in the repeated
measurements of level 2, which was used as diluent instead of PBS (the VetScan cannot process
PBS as was used for all other instruments).
All of the in-clinic hematology analyzers required minimal maintenance and minimal hands-
on time (except the QBC) to process samples. However, there was still a need for adequate
training to ensure proper sample processing and instrument maintenance as well as to know
how to handle flagged samples. The long cycle time of the LaserCyte can be a disadvantage
in clinics processing more than 4 samples per hour. However, the user can perform other tasks
while the instrument is analyzing a sample.
While erythroid regeneration can be estimated from the amount of polychromasia in a routinely
stained blood film, a reticulocyte count is the best and most objective parameter of bone marrow
responsiveness in dogs and cats. However, microscopic enumeration of reticulocytes is
laborious and time-consuming. Automated reticulocyte counts, now readily available on the
LaserCyte and ForCyte instruments, reduce the workload in clinical laboratories and also
ensure more accurate classification and monitoring of anemia. In our study, reticulocyte counts
on both in-clinic instruments had a clear negative bias (strong negative proportional error) in
comparison with the ADVIA. Thus, applying the ADVIA reference limits for reticulocyte
counts obtained on other analyzers may result in false classification of some anemias. The
reason for the proportional negative bias remains unclear, but it was corroborated by another
study in which a proportional negative bias was found for canine reticulocyte counts on the
LaserCyte in comparison to manual counts (unpublished data from our laboratory). In that
study, the ADVIA had a negligible bias for canine reticulocyte counts in comparison to the
manual count. At present, the LaserCyte and ForCyte can determine reticulocyte counts in both
canine and feline samples; however, at the time of data collection, only the LaserCyte provided
feline reticulocyte counts.
In conclusion, assessment of accuracy and precision of CBC data is hampered by the lack of
an accurate and rapid reference method, universal control reagents, and the complexity and
instability of blood cells. Nevertheless, for most cell counts and RBC parameters, the in-clinic
analyzers evaluated in this study performed similarly and as well as their laboratory
counterparts. Undoubtedly, instrument and software technologies will continue to improve for
existing and new hematology instruments, making automated CBC results more accurate and
valuable for clinical veterinary practice. As these improvements are made, comprehensive
correlation studies with acceptable reference laboratory technologies are both expected and
needed.
Supplementary Material
Refer to Web version on PubMed Central for supplementary material.
Acknowledgments
The authors thank the laboratory staff of Clinical Diagnostic Solutions, Inc., Plantation, Florida, for their technical
support. The following clinicians and veterinary clinics in southern Florida providing the blood samples are also
acknowledged: Dr. L. Dee at Hollywood Animal Hospital; Dr. R. Seaman at Coral Springs, and Dr. R. Buzzetti at
Imperial Point Animal Hospital. The study was supported in part by IDEXX Laboratories Inc., Westbrook, ME.
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14. Schwendenwein I, Jolly M. Automated differentials by an impedance on-site hematology system

[abstract]. Vet Clin Pathol 2001;30:158.
15. Pinches MDG, Burley K, Cue SM, Crawford EM, Papasouliotis K. Comparison of white blood cell
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Saunders; 2001. p. 245
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27. Matthews JA, Jensen M, DeNicola DB. Comparison of standard haematology parameters determined
on a laser-based point-of-care haematology analyser and an automated haematology analyser
commonly used in veterinary reference laboratories. J Vet Intern Med 2003;17:383.
28. Weissert, D.; Becker, M.; Moritz, A. Anämieklassifikation bei Hunden mittels automatisierter
durchflusszytometrischer Retikulozytenanalyse. Proceedings of the 16th Jahrestagung der
Fachgruppe “Innere Medizin und Klinische Labordiagnostik” der DVG; Giessen, Germany.
Tieraerztliche Praxis; 2008. p. A11-A12.
Figure 1.
Bland-Altman plots for analytes and instruments with high numbers of flagged samples,
differentiating between flagged (○) and unflagged (●) samples. The graph on the upper left
shows WBC counts from the CELL-DYN 3500 for canine samples. Discrepant results are
flagged. The graph on the upper right shows WBC counts from the CELL-DYN for feline
samples. Most discrepant results are flagged. The graph on the lower left shows PLT counts
from the Scil Vet ABC for feline samples. Many discrepant results, as well as many accurate
results, are flagged. The graph on the lower right side shows PLT counts from the QBC
VetAutoread for canine samples. Many discrepant results, as well as many accurate results,
are flagged.
NIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author Manuscript
Table 1
Specifications of the in-clinic and reference hematology instruments evaluated in this study.
Manufacturer Technology Reticulocyte Count Sample Volume (µL) Analysis Time (min) Control Material Internal Data Storage Software Version
ents
Becker et al.
IDEXX Laboratories, Inc., Laser Canine/feline 95 12 Latex beads Unlimited 1.37/1.1.87

Westbrook, ME, USA
Oxford Science, Inc., Laser and impedance Canine* 20 2 NA NA NA
Oxford, CT, USA
Melet Schloesing Impedance No 16 3 NA 100 samples 5.04 A
Pharmaceuticals, LaChaux
de Fonds, Switzerland
Boule Medicalab, Impedance No 125 1 Blood 250 samples V3.82s-r
Stockholm, Sweden
Scil Animal Care Co., Impedance No 12 1.5 Blood 60 samples NA
Viernheim, Germany
Abaxis, Sunnyvale, CA, Impedance No 10–20 2 Blood 100 samples 3.51 U
USA
read IDEXX Laboratories, Inc., Buffy coat No 111 7† Calibration rod No 4.3
Abbott Laboratories, Laser and Impedance No ~130 0.5 Blood Unlimited Rev.H Ver 4.3
L- Abbott Park, IL, USA
Siemens Medical Optical and peroxidase Canine/feline 157 1 Blood Unlimited V2.2.2A
Solutions, Fernwald,
Germany
nt to this study also include feline reticulocyte counts.
sample preparation time.
at time of study.
Page 13
Table 2
Coefficients of variation (%) in the replication experiment.
Analyte LaserCyte ForCyte MS45 Heska CBC Scil Vet ABC VetScan CELL-DYN
WBC 3.1 2.3 3.2 2.6 2.2 1.4 1.4

Becker et al.
RBC 4.1 2.8 1.2 2.0 2.7 1.4 0.5

HGB 2.9 1.3 1.1 2.3 1.8 0.7 0.6
HCT 3.9 2.8 1.6 2.5 2.7 2.1 0.7
MCV 0.4 1.0 0.9 0.7 0.6 1.0 0.6
PLT 4.7 4.4 6.7 6.4 7.2 9.2 2.2
Page 14
Table 3
Percentage of flagged measurements in samples from dogs and cats.
LaserCyte ForCyte MS45 Heska CBC Scil Vet ABC VetScan QBC VetAutoread CELL-DYN ADVIA
Analyte Dog Cat Dog Cat Dog Cat Dog Cat Dog Cat Dog Cat Dog Cat Dog Cat Dog Cat
Becker et al.
N 260 109 90 44 178 72 242 95 249 107 236 97 173 50 259 110 224 87
WBC 1 3 8 20 3 14 1 0 6 22 2 5 21 28 44 72 80 66
RBC 1 4 8 20 3 3 0 2 1 3 2 3 — — 2 4 79 74
HGB 1 2 8 20 1 3 0 0 1 0 1 1 9 40 2 4 33 59
MCV 0 0 8 20 1 3 0 0 0 0 1 1 — — 2 4 79 67
HCT 0 0 8 20 1 3 0 0 1 0 1 1 0 0 2 4 33 59
PLT 1 3 8 20 3 1 3 89 4 44 3 2 57 68 2 4 80 56
Page 15
Table 4
Results of comparing in-clinic instruments and the CELL-DYN to the ADVIA 120 reference standard.
e, Species Parameter LaserCyte ForCyte MS45 Heska CBC Scil Vet ABC VetScan QBC VetAutoread CELL-DYN
anine N 209 70 141 203 204 189 136 204

Becker et al.
r 0.990 0.992 0.989 0.990 0.988 0.992 0.942 0.994

Sy|x 1.55 1.74 1.78 1.10 1.11 1.53 3.62 1.27
y = ax+b 1.04x−0.26 1.03x+0.29 1.03x+0.65 1.03x−0.39 1.13x−0.21 1.03x+0.72 0.97x+2.06 1.01x−0.79

Bias, 109/L 0.08 0.81 1.10 0.99 1.11 1.21 1.61 −0.58
95% limits −2.95 to 3.10 −2.61 to 4.22 −2.41 to 4.61 0.82 to 1.19 0.91 to 1.35 − 1.82 to 4.23 −5.72 to 8.93 −3.09 to 1.94
eline N 74 26 50 67 76 69 33 74
r 0.986 0.966 0.970 0.993 0.979 0.984 0.950 0.992
Sy|x 1.56 2.50 2.19 1.20 2.07 1.67 3.08 1.16
Y = ax+b 1.01x−0.14 1.05x−0.34 1 .00x+0.45 1.04x−0.52 1.10x−1.20 1.05x+0.11 1.22x−0.74 0.99x−0.37

Bias, 109/L −0.15 0.54 0.51 −0.11 0.22 0.75 2.42 −0.49
95% limits −3.18 to 2.88 −4.27 to 5.36 −3.73 to 4.75 − 2.57 to 2.53 − 4.03 to 4.47 −2.53 to 4.04 −3.90 to 8.74 −2.77 to 1.80
anine N 213 74 142 201 202 186 — 208

r 0.983 0.965 0.983 0.990 0.992 0.985 — 0.985
Sy|x 0.31 0.48 0.29 0.26 0.22 0.25 — 0.27
Y = ax+b 1.02x−0.05 1.01x+0.34 1.00x+0.17 1.07x+0.06 1.02x+0.08 0.90x+0.55 — 1.00x+0.01

Bias, 1012/L 0.04 0.41 0.17 0.44 0.22 −0.04 — 0.00
95% limits − 0.57 to 0.65 −0.51 to 1.34 −0.39 to 0.73 −0.10 to 0.98 −0.21 to 0.66 − 0.64 to 0.56 −0.52 to 0.53
eline N 73 30 51 68 76 70 — 76
r 0.981 0.899 0.971 0.985 0.983 0.975 — 0.979
Sy|x 0.42 0.82 0.58 0.40 0.44 0.46 — 0.44
Y = ax+b 1 .00x+0.03 1.00x+0.74 1 .00x+0.56 1.07x+0.41 1.09x+0.27 0.90x+0.78 — 1.06x+0.13

12
Bias, 10 /L 0.01 0.75 0.55 0.89 0.85 0.11 — 0.52
95% limits − 0.80 to 0.82 −0.83 to 2.33 −0.59 to 1.68 0.08 to 1.70 − 0.07 to 1.77 −0.91 to 1.12 −0.35 to 1.38
anine N 208 71 141 201 202 184 144 206

r 0.988 0.987 0.978 0.990 0.994 0.993 0.983 0.995
Page 16
Sy|x 0.65 0.67 0.78 0.60 0.47 0.46 0.74 0.38
Y = ax+b 1.05x−0.68 0.95x+0.41 0.96x−0.87 1.03x−0.08 1.05x−0.27 0.97x+0.79 0.95x−0.55 0.97x+0.17

Bias, g/dL 0.13 −0.21 −1.49 0.40 0.47 0.33 −1.33 −0.30
Becker et al.
95% limits −1.17 to 1.43 − 1.58 to 1.61 −3.07 to 0.08 −0.80 to 1.61 − 0.51 to 1.45 −0.60 to 1.27 −2.85 to 0.19 −1.09 to 0.49
eline N 75 27 50 65 74 67 34 73
r 0.983 0.920 0.973 0.989 0.987 0.989 0.983 0.987
Sy|x 0.63 0.73 0.83 0.52 0.58 0.53 0.57 0.50
Y = ax+b 0.99x+0.16 1.03x−0.09 1.00x − 1.14 1.06x+0.10 1.05x−0.53 1.05x+0.19 0.93x−0.50 0.99x+0.16
Bias, g/dL 0.10 0.25 −1.19 0.74 0.07 0.73 −1.28 0.08
95% limits −1.12 to 1.33 −1.14 to 1.65 −2.80 to 0.43 − 0.32 to 1.80 −1.11 to 1.25 −0.32 to 1.79 −2.49 to −0.08 −0.89 to 1.06
anine N 209 73 140 199 201 184 — 207

r 0.813 0.879 0.820 0.893 0.920 0.849 — 0.938
Sy|x 2.84 2.26 0.67 2.32 1.82 3.40 — 1.74
y = ax+b 1.15x—10.15 0.97x−3.06 0.88x+9.42 1.02x−4.77 0.89x+4.27 1.22x−8.13 — 1.03x−0.37

Bias, fL 0.65 −4.29 1.01 −3.53 −3.33 7.76 — 1.87
95% limits − 5.25 to 6.56 −9.56 to −0.27 −5.29 to 7.31 − 8.25 to 1.20 −7.47 to 0.80 1.10 to 14.41 −1.60 to 5.34
eline N 72 30 50 68 76 69 — 71
r 0.759 0.901 0.947 0.937 0.946 0.948 — 0.953
Sy|x 2.20 1.65 1.84 1.95 1.64 2.45 — 2.28
y = ax+b 0.93x+2.46 0.88x+3.63 0.82x+7.85 0.94x+0.13 0.88x+3.44 1.26x−4.31 — 1.25x−8.41
Bias, ft. −1.15 −2.48 −1.22 −2.57 −2.31 8.29 — 3.69
95% limits −6.80 to 4.49 −6.14to 1.17 −5.87 to 3.43 −6.66 to 1.51 −6.13 to 1.52 3.05 to 13.53 −1.12 to 8.49
anine N 204 69 135 190 195 169 124 200

r 0.886 0.790 0.870 0.887 0.890 0.804 0.789 0.824
Sy|x 73.13 103.80 75.46 66.12 57.27 79.99 101.2 76.71
y = ax+b 1.08x−17.02 0.97x+15.60 1.05x−20.00 0.97x−14.48 0.84x+1.06 1.08x−19.74 1.13x−17.46 0.98x−24.26

Bias, 109/L 8.38 6.38 −3.64 −22.19 −47.93 −8.02 20.31 −31.65
Page 17
95% limits −134.93 to 151.69 −198.08 to 210.83 −151.24 to 143.96 −152.03 to 107.65 −168.65 to 72.79 −171.95 to 155.92 −177.58 to 218.19 −188.60 to 125.31
eline N 76 30 50 67 74 69 34 75
Becker et al.
r 0.883 0.837 0.779 0.716 0.767 0.685 0.847 0.766

Sy|x 100.4 147.5 103.7 117.1 91.23 96.65 83.58 105.3
y = ax+b 1.18x−27.51 1.30x−24.76 0.97x+54.88 0.80x+54.98 0.72x+38.16 0.59x+47.10 1.05x−1.21 1.05x−46.74

9
Bias, 10 /L 22.98 77.20 45.44 −1.04 −44.71 − 69.33 11.88 −32.67
95% limits −174.95 to 220.91 −214.19 to 368.59 −156.48 to 247.36 −240.39 to 238.30 −243.66 to 154.23 −294.05 to 155.39 −150.05 to 173.82 −244.75 to 179.41
ces in sample number are due to exclusion of samples with insufficient volume or with error messages indicating instrument malfunction.
Page 18
Table 5
Results of comparing in-clinic instruments and laboratory hematology analyzers to PCV as the reference standard.
Species Parameter LaserCyte ForCyte MS45 Heska CBC Scil Vet ABC VetScan QBC VetAutoread CELL-DYN ADVIA
Canine N 249 80 177 233 239 234 162 245 202

Becker et al.
r 0.957 0.948 0.959 0.979 0.978 0.975 0.990 0.971 0.982

Sy|x 3.56 4.20 3.35 2.54 2.48 2.79 1.71 2.70 2.19
y = ax+b 1.05x−0.06 1.03x−0.01 1.05x+1.34 1.05x+0.25 1.03x−0.54 1.09x+2.11 1.02x−0.95 1.03x+0.80 1.04x − 0.38
Bias, % 1.93 1.25 3.27 2.23 0.83 5.73 0.03 2.29 1.09
95% limits −5.09 to 8.95 −6.29 to 9.42 −3.29 to 9.83 −2.48 to 7.30 −4.03 to 5.69 0.12 to 11.35 −3.22 to 3.38 −3.01 to 7.58 −3.21 to 5.40
Feline N 100 41 70 88 101 96 50 99 75

r 0.943 0.887 0.957 0.975 0.969 0.927 0.991 0.950 0.981
Sy|x 3.15 3.46 3.14 2.18 2.49 4.12 1.32 3.20 1.90
y = ax+b 0.98x+1.51 0.89x+6.62 1.06x+1.56 1.05x+1.89 1.05x+2.15 1.13x+2.99 1.05x−0.61 1.14x+1.98 1.06x−0.39
Bias, % 0.74 3.21 3.43 3.50 3.82 7.18 1.01 6.61 1.37
95% limits −5.43 to 6.91 −3.88 to 10.32 −2.71 to 9.57 −0.83 to 7.82 −1.10 to 8.73 −0.89 to 15.25 −1.67 to 3.69 0.24 to 12.98 −2.38 to 5.12
Differences in sample number are due to exclusion of samples with insufficient volume or with error messages indicating instrument malfunction.
Page 19

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Vet Clin Pathol. 2008 December ; 37(4): 373–384. doi:10.1111/j.1939-165X.2008.00085.x.

Comparative clinical study of canine and feline total blood cell

Martina Becker1, Andreas Moritz1, and Urs Giger2

Justus-Liebig University Giessen, Giessen, Germany

© 2008 American Society for Veterinary Clinical Pathology

Materials and Methods

Linearity, precision, and carry-over

The International Council for Standardization in Hematology (ICSH) defines accuracy as “a

intervals are recommended to avoid clinically relevant misinterpretations. Of more concern

Pract 2001;42:326–332. [PubMed: 11480897]

14. Schwendenwein I, Jolly M. Automated differentials by an impedance on-site hematology system

IDEXX Laboratories, Inc., Laser Canine/feline 95 12 Latex beads Unlimited 1.37/1.1.87

nt to this study also include feline reticulocyte counts.

sample preparation time.

WBC 3.1 2.3 3.2 2.6 2.2 1.4 1.4

RBC 4.1 2.8 1.2 2.0 2.7 1.4 0.5

anine N 209 70 141 203 204 189 136 204

r 0.990 0.992 0.989 0.990 0.988 0.992 0.942 0.994

y = ax+b 1.04x−0.26 1.03x+0.29 1.03x+0.65 1.03x−0.39 1.13x−0.21 1.03x+0.72 0.97x+2.06 1.01x−0.79

Y = ax+b 1.01x−0.14 1.05x−0.34 1 .00x+0.45 1.04x−0.52 1.10x−1.20 1.05x+0.11 1.22x−0.74 0.99x−0.37

anine N 213 74 142 201 202 186 — 208

Y = ax+b 1.02x−0.05 1.01x+0.34 1.00x+0.17 1.07x+0.06 1.02x+0.08 0.90x+0.55 — 1.00x+0.01

Y = ax+b 1 .00x+0.03 1.00x+0.74 1 .00x+0.56 1.07x+0.41 1.09x+0.27 0.90x+0.78 — 1.06x+0.13

anine N 208 71 141 201 202 184 144 206

Sy|x 0.65 0.67 0.78 0.60 0.47 0.46 0.74 0.38

Y = ax+b 1.05x−0.68 0.95x+0.41 0.96x−0.87 1.03x−0.08 1.05x−0.27 0.97x+0.79 0.95x−0.55 0.97x+0.17

anine N 209 73 140 199 201 184 — 207

y = ax+b 1.15x—10.15 0.97x−3.06 0.88x+9.42 1.02x−4.77 0.89x+4.27 1.22x−8.13 — 1.03x−0.37

anine N 204 69 135 190 195 169 124 200

y = ax+b 1.08x−17.02 0.97x+15.60 1.05x−20.00 0.97x−14.48 0.84x+1.06 1.08x−19.74 1.13x−17.46 0.98x−24.26

r 0.883 0.837 0.779 0.716 0.767 0.685 0.847 0.766

y = ax+b 1.18x−27.51 1.30x−24.76 0.97x+54.88 0.80x+54.98 0.72x+38.16 0.59x+47.10 1.05x−1.21 1.05x−46.74

Canine N 249 80 177 233 239 234 162 245 202

r 0.957 0.948 0.959 0.979 0.978 0.975 0.990 0.971 0.982

Feline N 100 41 70 88 101 96 50 99 75

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