Beruflich Dokumente
Kultur Dokumente
A flow cytometer for the automated analysis of urinary sediment was designed, and its performance was
examined by the evaluation of 821 specimens. Auramine 0, a dye for DNA and RNA, was used for the staining
of the sediment. Urine (5 ml or more) was processed by the instrument for sediment analysis. Conventional
microscopic analysis was done for comparison. The RBC count, the WBC count, and the number of bacterial
cells, epithelial cells, and casts found by the flow cytometer and by microscopy were compared. Correlation
was high for all these results. The overall sensitivity, specificity, and efficiency (accuracy) in the items
analyzed were 84.7%, 57.8%, and 67.2%, respectively. One hundred specimens could be analyzed by the
instrument per hour. The instrument seemed useful for screening for urinary tract disorders to identify
specimens that should be analyzed microscopically in routine laboratories. o 1995 Wiley-Liss, Inc.
Key terms: Urine, sediment, flow cytometry, laboratories, hospital, autoanalysis, urinary tract disorders
Microscopic Analysis
Ten milliliters of fresh urine was centrifuged at 500
X g for 5 min, and its supernatant was removed by aspiration, leaving 0.2 ml of urine, which contained the sediment. After gentle mixing of the sediment, it was
smeared onto a slide and observed under a light microscope at 400 X (high power field; HPF) except for casts.
Casts were observed at 100X (low power field; LPF)
(3,4).
Statistics
Statistical analysis was done with Pearsons method for
correlation studies and with Wilcoxons test for nonparametric group studies (2). Differences for which P was
less than 0.005 were taken to be significant.
Efficiency Test
Instrument
A flow cytometer used for reticulocyte analysis (Sysmex R-1000, Toa Medical Electronics Co., Kobe, Japan)
was modified for urinary sediment analysis by the addition of devices for urine transport and software for signal
analysis, both designed with the technical cooperation of
the manufacturer. In testing, a sample cup containing
0 1 9 9 5 Wiley-Liss, Inc.
76
YASUI ET AL.
Forward scatter signal
REC
Fluorescencesignal
Epithelial cell
Bacteria
FIG. 1. Forward scatter signals and fluorescence signals of components of urinary sediment.
sults were true positives was judged by microscopic analysis of the urinary sediment.
RESULTS
Scattergrams of Components of Sediment
Figure 2 shows a negative sample and a positive sample
with all five main components (REK, WBC, bacteria, epithelial cells, and casts). The positive sample shows typical patterns when each of the five components, which
may be found in abnormal urinary sediment, were increased to more than that of a negative specimen. All
types of epithelial cells, such as squamous cells, transi-
Positive Sample
Negative Sample
Casts
_-CL
P I
LL
FIG.2. Scattergrams of negative sample and positive sample with all components. FLS, forward light scatter;
FI, fluorescence intensity; NL, nuclear length; CL, cell length.
77
R BC
Table 1
Reproducibility Test (Performed 10 Times)
Sample
Mean(/$)
SD
3.3
6.1
1098.8
4.1
97.2
1.0
0.6
20.5
1.4
6.5
29.9
9.5
1.9
34.1
6.7
34.9
18.9
87.5
16.5
72.5
4.4
5.0
4.7
1.8
4.1
12.6
26.6
5.4
11.1
5.7
43.9
276.0
149.1
105.4
276.8
4.1
16.3
3.8
8.2
13.1
9.3
5.9
2.6
7.8
4.7
37.2
31.3
19.5
15.9
21.3
2.7
4.4
2.4
1.5
1.7
7.3
14.1
12.2
9.7
8.1
0.05
0.05
0.19
0.10
0.10
29.1
61.0
55.8
94.4
29.1
RBC
1
2
3
4
5
WBC
1
2
3
4
5
y = 0.363~
+ 0.946. r = 0.765, P ~0.0001
CN%)
Epithelial cells
3
4
5
Casts
1
2
3
4
200
400
(MPF)
Microscopy
WBC
Bacteria
1
2
3
4
5
0.04
0.03
0.34
0.05
Microscopy
Epithelial cells
y = 0.308~
+ 0.638, r = 0.619, P ~0.0001
20
40 (MPF)
Microscopy
Renal Disorders
The numbers of epithelial cells and casts in patients
with renal disorders were significantly higher than in the
other subjects (Table 5). Renal disorders were judged
from the total results after analysis of urinary sediment.
Speed
The automated method can analyze 100 specimenshr.
DISCUSSION
A medical technologist first scans the whole field by
78
YASUI ET AL
Table 2
Significant Differences in Values Between Sexes for Five Items
RBC
WBC
Bacteriab
Epithelial cells
Casts
Total
(/H PF)
9.0 f 70.0
7.3 f 55.6
0.5 f 1.5
5.0 t 15.8
2.0 i_ 23.1
Microscopy
Men
Women
(IHPF)"
(IHPF)a
10.6 t 7 9 . 2
7.2 t 56.9
10.0 2 70.3
4.0 2 27.3
0.2 2 0.5
0.7 t 1.6
1.9 +- 5.8
7.6k19.1
2.9 k 31.6'
1.3 s 12.3'
P
0.0004
<0.0001
<0.0001
<0.0001
0.7005
Automated
Men
(Iul)a
3 . 1 t 29.7
2.8
15.0
0.7 t 2.1
0.7k2.1
0.5 k 4.7
Total
Uul)
4.1 k 3 1 . 8
4.9
42.2
0.9 2.3
2.2t7.8
0.4 k 3.8
*
*
analysis
Women
Uul)
4.6 t 32.4
54.9
6.6
1.1 f 2 . 6
1.122.6
0.5 2 3.2
<0.0001
<0.0001
<0.0001
<0.0001
0.0056
Table 4
Overall Efficiency of Sediment Analysis of Five Items"
Table 3
Results of Evaluation of Automated Analysis"
Sensitivity (Yo)
Specificity (%)
Efficiencv (Yo)
RBC
68.5
76.1
74.6
WBC
84.9
90.7
90.1
Bacteria
65.8
85.2
83.3
Epithelial
cells
74.5
89.0
86.3
Casts
61.7
72.7
71.5
Positive
sarnDlesC
222
206
428
Positive sa m piesb
Negative Sam plesb
Total samdes
Negative
SamDles'
40
282
322
Total
samoles
262
488
750
Table 5
Results i n Patients with Renal Disorders
RBC
WBC
Bacteriab
Epithelial cells
Casts
Renal disordersa
(/HPF)
13.86 I109.27
7.76 2 38.83
0 . 6 3 -t 1.44
6.17 t 16.92
4.46 ? 36.68'
Microscopy
Othera
(IHPF)
8.42 t 62.8
7.14 t 57.69
0.50 2 1.46
4.79 f 15.39
1.69
20.46'
Automated analysis
Renal disordersa
Othera
P
0.2412
0.9637
0.0365
0.0290
0.0016
(/PI)
4.39
5.41
1.05
2.98
0.92
k
k
k
k
k
35.28
26.28
2.63
8.57
7.57
(IpC
3.90 t 30.75
4.70 k 43.81
0.90 ? 2 . 3 4
2.06 k 7.66
0.40 +- 3.08
P
0.0269
0.1998
0.3937
0.0081
0.0357
ACKNOWLEDGMENTS
We thank Dr. Mikio Okamura, First Department of Internal Medicine, Osaka City University Medical School,
for providing information about his patients with kidney
disease.
LITERATURE CITED
1. Davis BH, Bigelow NC: Clinical flow cytometric reticulocyte analysis. Pathobiology 58:99-106, 1990.
79