52
Test 1 Ed. 1, Rev 2 25.01.10
Test 1
Ed. 1, Rev 2 25.01.10
Test 1
Test 1 technology
Test 1 Roller 20
NEW
Test 1
Test 1
ESR: distance in millimeters covered by the
erythrocytes in non coagulated blood into a
dedicated tube set in vertical position,
perpendicular to an horizontal surface in 60
minutes.
It is a non specific test for the screening of
different pathologies.
ESR is a time limited physical phenomenon
with poor reproducibility
Erythrocyte Sedimentation Rate
Test 1
Erythrocyte Sedimentation Rate
Classical Westergren/Wintrobe method (1921)
Test 1
Metodo classico di Westergren (1921)
T=0
T=0
Healthy subject
Patient under
inflammatory
state
T=1 hour
T=1 hour
…red cells are at the
same level
…red cells follow
Normal and patological contition
Test 1
Test1 Features
▪ Fully automated ESR system
▪ Engineered to respect CLSI requirements
▪ High correlation with standard Westergren method
▪ Total capacity up to 60 samples with a production
up to180 samples per hour
▪ 150 µl blood per sample in EDTA
▪ First result after 3 minutes
▪ Results in 20 seconds
▪ Thermostated at 37°C
▪ Complete integration with Haematology Lab
▪ Use of the same CBC EDTA samples
▪ 6 different models: 5 for CBC rack direct loading
▪ Internal barcode reader
▪ Latex Calibrators & Quality Controls
▪ Statistical Control
▪ Smart Card system
▪ Thermal printer
▪ Connection with LIS
results in 20 seconds
PATENT TECHNOLOGY
Test 1
Test 1 technology advantages
Results in 20 seconds related to red cells aggregation
kinetics
High correlation with Westergren
EDTA used as anticoagulant
Thermostated at 37°C
No influence by low hematocrit level (≤ 0.35)
Fixed micro-volume of 150 micro-liters per sample
180 samples/hour
Mixing for complete inversion
Sample traceability
Quality Controls
Statistical Qualitu control
Low waste production
Test 1
CBC Racks Direct Loading
5 Test1 models for direct loading:
Test 1 BCL SI 195.220 BCL
Test 1 SDL SI 195.230 SDL
Test 1 YDL SI 195.240 YDL
Test 1 MDL SI 195.250 MDL
▪ To obtain the complete integration in
the haematology lab now it’s possible
to load the same CBC racks on Test1
saving time and possible operator
mistakes.
▪ Direct loading is available for Beckman
Coulter, Siemens and Sysmex cell
blood counters
▪ Traceability of the sample is granted
by the Internal Barcode Reader!
▪ Total capacity up to 60 samples with
a production up to 180 samples per
hour
Test 1
Test1 family Smart Card system
▪ All Alifax instruments do not require
special consumable reagents.
▪ Instruments are enabled to work with
dedicated smart cards where the ESR
tests are loaded.
▪ Smart cards are available in different
size
▪ Environmentally friendly cards save
storage and transport costs
for 10.000 TESTS of
for 20.000 TESTS of
Test 1
The TEST1 technology is
PATENTED
Test 1
TEST 1 Technology
TEST 1 analyses the Aggregation Capacity of the
blood red cells via Optical Density reducing the
analysis time to only 20 seconds!!!
Aggregation
Starting (T = 0)
Aggregation after
20 sec
Light beambefore
Light beamafter
Every sample is read 1000 times in 20 seconds
--
-
- -- -
+
--
+ +
+ ++
+
+
+ +
+
+
-+
+
+
+++
+
+
-
--
-
--
PATENT TECHNOLOGY
Test 1
TEST 1 Technology
Every sample is read 1000
times in 20 seconds
PATENT TECHNOLOGY
Test 1
ESR Phases
1a phase: aggregation
Erythrocytes start forming firsts rouleaux;
sedimentation begins
2a phase : precipitation
Erythrocytes-plasmatic proteins interface is
very rapid; sedimentation phase continues
3a phase : packaging
Erythrocytes aggregate and package to the
tube bottom
Test 1 Typical Sigmoid Curve of Red cells sedimentation in a
inflammatory state
Roleaux Formation
Precipitation
Packaging
Se
dim
e
nta
tio
n
(mm
)
0
200
0 Time
(minutes)
60
-+
++
+++
++
-
--
-
--
-++
++++
++++++
++++
-
--
-
--
-+
++
+++
++
-
--
-
--
-++
++++
++++++
++++
-
--
-
--
-+
++
+++
++
-
--
-
--
-++
++++
++++++
++++
-
--
-
--
-
-
-
- -- -
+
--
+ +
+ ++
+
+
+ +
+
+
-
-
-
- -- -
++
--
++ ++
++ ++++
++
++
++ ++
++
++
Test 1
1st Phase: Aggregation
T = 0
Aggregation starting
Roleaux formation
0
200
Agglomerin
Erythrocytes start forming firsts rouleaux; sedimentation begins
-
-
-
- -- -
+
--
+ +
+ ++
+
+
+ +
+
+
Test 1
2nd Phase: Precipitation
Plasma
0
200Roleaux
Erythrocytes-plasmatic proteins interface is very rapid;
sedimentation phase continues
-+
++
+++
++
-
--
-
--
-+
++
+++
++
-
--
-
--
-+
++
+++
++
-
--
-
--
Test 1
3rd Phase: Packaging
T = 60 min
Final reading point
0
200
Plasma
Roleaux packaging
Erythrocytes aggregate and package to the tube bottom
Test 1
Reading Cell – i.e. pathologic sample
1 µl
T=0 T= 20 sec
Test 1
1 µl
T=0 T= 20 sec
Reading Cell – i.e. healthy sample
Test 1
In pathological condition the blood undergoes
changes and the concentration of acute phase
proteins as fibrinogen, IgM, α-macroglobulin,
called AGGLOMERINS, increases in plasma.
Rouleaux start to precipitate
Different surface electric
charges of Erythrocytes (-
) and plasma proteins (+)
interact causing red cells
aggregates called
Rouleaux.
Test 1
Inflammation Status
Roleaux Formation
Precipitation
Packaging
Se
dim
en
tatio
n
(mm
)0
200
0 Time (minutes) 60
Typical Sigmoid Curve
-+
++
+++
++
-
--
-
--
-++
++++
++++++
++++
-
--
-
--
-+
++
+++
++
-
--
-
--
-++
++++
++++++
++++
-
--
-
--
-+
++
+++
++
-
--
-
--
-++
++++
++++++
++++
-
--
-
--
-
-
-
- -- -
+
--
+ +
+ ++
+
+
+ +
+
+
-
-
-
- -- -
++
--
++ ++
++ ++++
++
++
++ ++
++
++
Test 1
Normal Status
Latency Phase
Time (minutes)
Se
dim
en
tatio
n
(mm
)0
0 60
No agglomerines
presence
- -
--- -
-
-
--
- -
-
--
- -
-
--
- -
-
--
- -
- -
--- -
-
--
- -
- -
--- -
-
-
--
- -
-
--
- -
-
--
- -
-
--
- -
- -
--- -
-
--
- -
Test 1
How TEST 1 Works
CPS at 37°C
Reading cell
(1 µl)
Sample 1, 150μlAir BubbleSample 2, 150μl
Test 1
PATENT OF TEST 1
The world patent of Test-1
concerns the mathematic
algorithm that expresses the same
sigmoid function of the ESR
Test 1
Correlation with WG
We have almost 100 publications that
demonstrate an excellent correlation of Test1
ESR results with the Westergren reference
method and commercial automated systems
based on sedimentation method
The results of different works demonstrate
correlation coefficients >94%-97%
Test 1
r2= 94%.
r2= 94%.
r2= 99%.
r2= 97%.
r2= 94%.
r2= 97%.
Correlation with WG - Publications
Erythrocyte sedimentation rate in blood a comparison of the Test 1 ESR system
with the ICSH reference method
J. Reis, J. Diamantino, N. Cunha, F. Valido (Clinical Pathology Department, IPO Coimbra; Francisco Gentil, EPE,
Portugal) Clinical Chemistry and Laboratory Medicine 2007 June; 45,
Comparison of TEST 1 with SRS 100 and ICSH reference method for the
measurement of the length of sedimentation reaction in blood
JS. Ozdem, H.S. Akbas, L. Donmez, M. Gultekin (Clinical Biochemistry Unit, Medical Faculty, Central Laboratory,
Akdeniz University, Antalya, Turkey) Clinical Chemistry and Laboratory Medicine. 2006;44(4):407-12
Determination of the Length of Sedimentation Reaction in Blood: a Comparison
of the Test1 ESR System with the ICSH Reference Method and the Sedisystem
A. Romero, M. Muñoz, G. Ramirez (Dept. of Haematology, H.C.U. "Virgen de la Victoria", Málaga & *GIEMSA, School
of Medicine, University of Málaga, Spain) Clinical Chemistry and Laboratory Medicine 2003, 41 (2).
Basic Evaluation and Reference Range Assessment of TEST1 for the
Automated Erythrocyte Sedimentation Rate
B.H. Lee, J. Choi, M.S. Gee, K.K. Lee, H. Park (Dept. of Laboratory Medicine, Kangbuk Samsung Hospital,
Sungkyunkwan University School of Medicine, Seoul, Korea) Journal of Clinical Pathology and Quality Control, Vol. 24,
No. 1, 2002.
Erythrocyte Sedimentation Rate. Use of Fresh Blood for Quality Control
M. Plebani, E. Piva (Dept. of Laboratory Medicine, University-Hospital, Padova, Italy) American Journal of Clinical
Pathology, 2002, 117:621-626.
Erythrocyte Sedimentation Rate by Test-1 Analyzer
N. de Jonge, I. Sewkaransing, J. Slinger, J.J.M. Rijsdijk (Dept. Clinical Chemistry, Leyenburg Hospital, The Netherlands)
Clinical Chemistry, June 2000, 46: 881-882.
Test 1
Test 1 respects the CLSI* requirements
1. Sample Mixing step
▪ Before the start analysis the instrument performs a mixing
cycle step for all the sample loaded
2. Temperature control
▪ The instrument is thermostated at 37°C
3. Material
▪ There is a single capillary in which the samples pass
through
4. Using of EDTA as anticoagulant
▪ EDTA is suggested as anticoagulant because increase the
stability and reduce the dilution errors caused by the using
of Sodium Citrate
*National Council for Clinical Laboratory Standardization:
“ Reference and Selected Procedure for ESR Test; Approved standard- 4th Edition”
vol.20 No.27; December 2000
Test 1
1. Temperature control
2. Sample intrinsic variation
3. Variation induced by delay in the workflow
4. Errors in the dilution step with Sodium Citrate
5. Low level of Hematocrit (≤0,35)
6. Inadequate materials
7. Inadequate quality controls
8. No calibration available
of Westergreen Method
Variables and Limitations
Test 1
Low hematocit influence
▪ Thanks to its unique technology Test1 results
don’t need to be corrected
▪ All the other systems present in the market
are affected by low hematocrit values that is a
crucial point in this test
Test 1 Different ESR Results of the same sample at different
hematocrit values
Sedimentation like time function at different hematocrit values artificially obtained adding
autologous plasma (Fabry T., Blood, Vol. 70, No 5, Nov. 1987: pp 1572-1576).
Hematocit 26 Hematocrit 28 Hematocrit 34 Hematocrit 45
SE
DIM
EN
TA
TIO
N
(mm
)
TIME (minutes)
0
20
40
60
80
100
120
140
0 20 40 60 80 100 120 140 160 180 200
Test 1
32
Low Hematocrit
▪ As well described in the scientific paper
written by TL Fabry* one reason for
variation in the ESR is the inverse
correlation to the hematocrit value.
▪ Fabry suggested a formula to adjust
the ESR considering the Hct value.
Hematocit 26 Hematocrit 28 Hematocrit 34 Hematocrit 45Hematocit 26 Hematocrit 28 Hematocrit 34 Hematocrit 45
SE
DIM
EN
TA
TIO
N
(mm
)
TIME (minutes)
0
20
40
60
80
100
120
140
0 20 40 6060 80 100 120 140 160 180 200
*Mechanism of Erythrocyte Aggregation and Sedimentation Blood, Vol. 70, No 5, Nov. 1987: pp 1572-1576
▪ None of the automated systems based on sedimentation ESR take
into account the hematocrit level.
Test 1
The temperature increases of 1 °C creates a variation of 3% of
viscosity.
Variation of the viscosity referring to the law of Stokes shows the change of
ESR results at different temperatures
Different Results of the Same Sample
20°C 25°C 37°C
24 ESR
28 ESR
38 ESR
Test 1
Test 1 measures the erythrocytes kinetics
aggregation in agglomerin presence. Thanks to
this unique characteristic is able to remove all
the variables and limitations of the classical WG
method based on the sedimentation.
All the other instruments simply read the final
result of a sedimentation, therefore present the
same limitations of the WG method.
Test 1
NCCLS suggests EDTA as anticoagulant that has to
be used to collect the sample that has to perform
the ESR test, because increases the blood stability.
NCCLS vol.20 No.27 “Reference and Selected Procedure for ESR Test; Approved standard-
4th Edition” page 6; chapter 8 “the ESR test: reference procedure ”
Test 1
THE EDTA COLLECTED SAMPLES
REFRIGERATED AT +4°C CAN BE
TESTED EVEN AFTER 24 HOURS
FROM BLOOD COLLECTION
From Clinical Chemistry, Vol. 47, No. 6, Supplement 2001, p. 162 - Internal quality
control for erythrocyte sedimentation rate (ESR) measured by TEST 1 analyzer by
D. Giavarina, S. Capuzzo, M. Carta, F. Caoduro, G. Soffiati, Clin. Chem. & Hematol Lab
- San Bortolo Hospital: Vicenza, Italia
Test 1
E. Heverin (Galway-Mayo Institute of Technology, Ireland), “Comparison of the Westergren method vs the TEST1 technique for determining the Erythrocyte Sedimentation Rate”, May 2002
K3EDTA @ 24hrs 4°C, K2EDTA @ 24hrs 4°C r=0.974
Reproducibility after 24h at 4°C
Test 1
K3EDTA @ 48hrs 4°C, K2EDTA @ 48hrs 4°C r=0.964
Reproducibility after 48h at 4°C
E. Heverin (Galway-Mayo Institute of Technology, Ireland), “Comparison of the Westergren
method vs the TEST1 technique for determining the Erythrocyte Sedimentation Rate”, May 2002
Test 1
E. Heverin (Galway-Mayo Institute of Technology, Ireland), “Comparison of the Westergren method vs the TEST1 technique for determining
the Erythrocyte Sedimentation Rate”, May 2002
ESR results at 4h versus 24 h at RT
Test 1
Test-1 ESR shows the aggregation activity even 12,
24, 48 hours after the samples collection (stored at
4°C), according to the sigmoid curve, that is the
unique curve described and represented for the ESR
by NCCLS (CLSI).
Conclusion
Test 1
Reproducibility with TEST 1
Thanks to the use of CPS thermostated at 37°C, Test1 shows the
stability of the assay (of 4%) independently of the external
temperature.
Y=0.9541
R2= 0.9923
0
10
20
3
0
40
50
60
20 40 60
13 10
50 454 42 22 2
23 2316 1412 103 36 7
54 536 6
10 112 29 92 22 25 69 85 6
23 2414 142 2
33 3112 114 4
15 16
22°C 26°C
Reproducibility with TEST 1 at 37°C
Test 1
Resutls – Repeatability and Reproducibility
Table 2a Repeatability results for a turbidimetric standard, lot n. 6 on one
TEST-1 SN 1077
Table 2b Reproducibility results for a turbidimetric standard, lot n. 6 on 3
different instruments TEST-1 SN 858, SN 860 and SN 1077 by 2 different
operators in 2 different periods of time (October and February)
Test 1
Internal statistical Quality control
* : HISTORICAL DATA
° : DAILY DATA
In this example historical
data are well aligned:
the system is working
properly.
Test 1
Sample Traceability
Traceability of the sample is granted by:
▪ the Internal or external Barcode Reader
▪ Using of the same CBC rack thus reducing
operator handling
▪ Alarm advisor of Missing barcode reading
Test 1
45
QC- Quality Control
1
Rev. 17.05.12
Do
cu
men
t13
Test 1
Latex Calibrators
▪ Thanks to latex technology we
can offer a useful calibration tool
to grant results stability and
reliability
▪ Using calibrators is possible to
easily align two or more systems!
For technical assistance use
Test 1
47
Latex Controls
Test 1
Turbidimetric standards
The kit is composed of 3 test tubes
each containing 3 ml of latex
solution at three different
concentration levels: Low, Medium,
High.
Low Medium High
The new latex control for the Test 1 family instrument is a
ready-to-use turbidimetric standard of synthetic latex
solutions
Each level has known turbidity values, on which the
instrument assesses photometric measurementscorrelated to ESR values.
Test 1
Latex Calibration
y = 0,9569x
R2 = 0,9783
0
10
20
30
40
50
60
70
0 20 40 60 80
Instrument 968
Instr
um
en
t 967
Latex Calibrators
Example of alignment of 2 instruments (Sn 968 and Sn
967) after calibration with Latex Calibrators
Test 1
External Quality Evaluation Latex Control
For inter laboratories
Quality Evaluation
Program
NEW
External Quality Evaluation Kit
For Test 1 family instrumentsSI 305.500 - SI 305.502 (Sarsted tubes)
1 box contains 3 ready to use tubes of unknown ESRlatex value.
Test 1
Test1 Maintenance
▪ Washing
▪ Check water values fluctuations
and trends
▪ Check the needles
▪ Check clots
▪ Check controls fluctuations and
trends
▪ Check harmonization
▪ Follow instructions
▪ Replace wash test tubes daily
▪ Call service support whenever
PATENT TECHNOLOGY
Test 1
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