Hematology Calibrator Traceability Report

Reference Measurement Laboratory of
Shenzhen Dymind Biotechnology Co., Ltd.
Hematology Calibrator
Traceability Research Report
Calibrator Model: DM-CAL PLUS
Calibrator Lot No./Exp. NK0123/2023-02-10

Date:
Written by: LIU Jia
Reviewed by: LIU Jinying
Approved by: ZHENG Jiantong
Release Date: 2022/12/28
Controlled or Not: ■ Yes □ No

File No.: RMLAB-WI-T-020
Technical File
Version: Version 1.0
Page: Page 2, 16 pages in total
1 Basis of Value Assignment

In accordance with the document guidance ISO 17511:2020, In vitro diagnostic medical devic-
es—Requirements for establishing metrological traceability of values assigned to calibrators,
trueness control materials and human samples.
2 Standard of Value Assignment

After querying the JCTLM database, Reference measurement procedure issued by the Interna-
tional Council for Standardization in Hematology（ICSH）and Standards issued by Clinical And
Laboratory Standards Institute（CLSI/NCCLS）, blood cell analysis components (RBC count,
WBC count, Determination of Hemoglobin, Determination of Packed Cell Volume and Platelet
Count) have primary reference measurement procedures but no certified primary reference
materials（CRM）. Although hemoglobin has a reference material, it is used as a trueness control
materials to test the accuracy of the instrument. Therefore, the traceability model of blood cell
analysis components is consistent. According to the ISO 17511:2020 document clause 5.3, the
measurement results can be traceable to SI. The traceability chain and calibration hierarchy of
blood cell analysis components are shown in Figure 1.
Material Measurement procedure
p.1.set of fit for purpose primary measurement pro-

cedures. e.g. counting, Spectrophotometry, length,
applied to primary measurement procedures[p.3] ap-
paratus and reagents for assurance of critical meas-
urement factors
p.3. primary measurement procedures defining the

measurand (WBC,RBC,HGB,HCT,PLT);full SI tracea-
m.4.Manufacturer’s bility (ISO 15193 conforming)
working calibrators
(Fresh blood sample)
p.5.Manufacturer’s standing measurement procedure
%rucalb m.5.End-user IVD MD

calibrators
p.6. End-user IVD MD
%ru(y)c m.6.Human sample

with result
u(y)
Figure 1 Metrological traceability chain of hematology analyzer’s quantity
Technical File
In Figure 1:
p.1: set of fit for purpose primary measurement procedure, the methods used for blood cell
analysis mainly include counting, spectrophotometer and length.
p.3: primary measurement procedures defining the measurand. Measured include the cell con-
centration of white blood cells, the cell concentration of red blood cells, the hemoglobin concen-
tration, the hematocrit, and the cell concentration of platelets in normal human fresh blood.
m.4: Manufacturer’s working calibrators. The reference measurement laboratory selects fresh
whole blood from healthy people as the experimental sample for the reference measurement
method, assigns it to the manufacturer's working calibrator.
p.5: Manufacturer’s standing measurement procedure. Hematology analyzers calibrated with the
manufacturer's working calibrator can be used as a standard machine after passing the verifica-
tion of performance tests, etc. The standard machine is routinely maintained in strict accordance
with the requirements, and the reliability of its results is ensured according to the standard ma-
chine management specifications.
m.5: End-user IVD MD calibrators End-user IVD medical device calibrators produced through a
qualified manufacturing process for the calibration of end-user instruments. The reference value
of the end-user in vitro diagnostic medical device calibrator is determined repeatedly by the
standard machine according to the corresponding assignment process, and the average value is
taken as the reference value.
p.6: End-user IVD MD. The assigned end-user IVD medical device calibrators can be used for
the calibration of end-of-line hematology analyzers. Calibrated hematology analyzers test rou-
tine samples with traceable results.
m.6: Human sample with result.
The specific traceability transmission chains involved in the blood cell reference measurement
procedure are as follows:
Technical File
2.1 White blood cell count (WBC):

p.1. set of fit for purpose primary measurement

procedures: counting
p.3. primary measurement procedures defining

the measurand (WBC): ICSH. Reference method
for the enumeration of erythrocytes and leuco-
m.4.Manufacturer’s cytes. Clinical and Laboratory Hematology,
working calibrators 16:131-138, 1994
(Fresh blood sam-
l )
p.5.Manufacturer’s standing measurement pro-
m.5. End-user IVD

%rucalb
MD calibrators
m.6. Human sample

%ru(y)c
with result
u(y)
Figure 2 Metrological traceability chain of WBC
WBC count refers to counting the number of white blood cells contained in a unit volume of
blood. The measurand is the amount of white blood cells in human fresh blood, and the type of
quantity is cell concentration and the unit of white blood cell count is /L. The matrix in the white
blood cell count is all components of fresh blood except white blood cells.
After consulting relevant international scientific/medical organizations (JCTLM, ICSH,
CLSI/NCCLS, etc.) and authoritative institutions (WHO, BCR, IRMM, NIST, etc.), WBC only has
the reference measurement procedure recommended by ICSH, and there is no reference mate-
rial. Therefore, we choose this reference measurement program as the reference measurement
program for leukocyte count. According to the ISO 17511:2020 document clause 5.3, the
traceability chain and calibration hierarchy of WBC is shown in Figure 2. p.3 is the primary
measurement procedures defining the measurand (WBC). m.4 is manufacturer’s working cali-
brators. Normal human fresh blood assigned by the reference method (p.3) is selected as the
manufacturer’s working calibrator. p.5 is manufacturer’s standing measurement procedure which
calibrated with the manufacturer's working calibrator. m.5 is end-user IVD MD calibrators.
End-user IVD MD calibrators assigned through a qualified manufacturing process for manufac-
turer’s standing measurement procedure. p.6 is End-user IVD MD. The assigned end-user IVD
medical device calibrators can be used for the calibration of end-of-line hematology analyzers.
Technical File
m.6 is human sample with result.
2.2 Red blood cell count (RBC)

p.1.set of fit for purpose primary measurement

procedures: counting

the measurand (WBC): ICSH. Reference method
for the enumeration of erythrocytes and leuco-
m.4.Manufacturer’s cytes. Clinical and Laboratory Hematology,
working calibrators 16:131-138, 1994
(Fresh blood sam-
ple)
%rucalb m.5.End-user IVD

MD calibrators
m.6.Human sample
%ru(y)c with result
u(y)
Figure 3 Metrological traceability chain of RBC
RBC count refers to counting the number of red blood cells contained in a unit volume of blood.
The measurand is the amount of red blood cells in human fresh blood, and the type of quantity is
cell concentration and the unit of white blood cell count is /L. The matrix in the red blood cell
count is all components of fresh blood except red blood cells.
CLSI/NCCLS, etc.) and authoritative institutions (WHO, BCR, IRMM, NIST, etc.), RBC only has
the reference measurement procedure recommended by ICSH (Reference method for the
enumeration of eryth-rocytes and leucocytes. Clinical and Laboratory Hematology, 16:131-138,
1994), and there is no reference material. Therefore, we choose this reference measurement
program as the reference measurement program for leukocyte count. According to the ISO
17511:2020 document clause 5.3, the traceability chain and calibration hierarchy of RBC is
shown in Figure 3. p.3 is the primary measurement procedures defining the measurand (RBC).
m.4 is manufacturer’s working calibrators. Normal human fresh blood assigned by the reference
method (p.3) is selected as the manufacturer’s working calibrator. p.5 is manufacturer’s standing
measurement procedure which calibrated with the manufacturer's working calibrator. m.5 is
Technical File
end-user IVD MD calibrators. End-user IVD MD calibrators assigned through a qualified manu-
facturing process for manufacturer’s standing measurement procedure. p.6 is End-user IVD MD.
The assigned end-user IVD medical device calibrators can be used for the calibration of
end-of-line hematology analyzers. m.6 is human sample with result.
2.3 Determination of Hemoglobin Concentration (HGB)

procedures: Spectrophotometry

the measurand (HGB): CLSI/NCCLS. H15-A3
Reference and Selected Procedure for the Quan-
titative Determination of Hemoglobin in
m.4.Manufacturer’s Blood.2000
working calibrators
(Fresh blood sam-
ple)
m.5.End-user IVD
%rucalb
MD calibrators

with result
u(y)
Figure 4 Metrological traceability chain of HGB
The hemoglobin concentration refers to the amount of hemoglobin contained in a unit volume (L)
of blood. The measurand is the hemoglobin concentration in human fresh blood, and the type of
quantity is mass concentration, and the hemoglobin concentration is expressed in g/L. The ma-
trix in the determination of hemoglobin is all components in fresh blood except hemoglobin in red
blood cells.
After consulting relevant international scientific/medical organizations (JCTLM, ICSH, CLSI/
NCCLS, etc.) and authoritative institutions (WHO, BCR, IRMM, NIST, etc.), HGB has the refer-
ence measurement procedure recommended by CLSI/NCCLS (standard H15-A3) and ICSH
(recommendations for reference method for hemoglobinometry in human blood (ICSH standard
1995) and specifications for international hemoglobincyanide standard (4th edition)). We se-
lected the latest version of the standard as the reference measurement procedure (standard
H15-A3). Hemoglobin has two reference materials, BCR 522 produced by European Commis-
sion and code: 98/708 produced by NIBSC. They are not primary reference materials, and their
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main function is used as a trueness control materials to test the accuracy of the instrument.
There is no difference in performance between them, but the reference material code 98/708 is
cheaper and easier to obtain, so we choose it as the reference material. According to the ISO
17511:2020 document clause 5.3, the traceability chain and calibration hierarchy of HGB is
shown in Figure 4. p.3 is the primary measurement procedures defining the measurand (HGB).
m.4 is manufacturer’s working calibrators. Normal human fresh blood assigned by the reference
method (p.3) is selected as the manufacturer’s working calibrator. p.5 is manufacturer’s standing
measurement procedure which calibrated with the manufacturer's working calibrator. m.5 is
end-user IVD MD calibrators. End-user IVD MD calibrators assigned through a qualified manu-
facturing process for manufacturer’s standing measurement procedure. p.6 is End-user IVD MD.
The assigned end-user IVD medical device calibrators can be used for the calibration of
end-of-line hematology analyzers. m.6 is human sample with result.
2.4 Hematocrit Determination (HCT)

procedures: Length

the measurand (HCT): CLSI/NCCLS. H7-A3
m.4.Manufacturer’s Procedure for Determining Packed Cell Volume
working calibrators by the Microhematocrit Method.2000
(Fresh blood sam-
ple)

MD calibrators

with result
u(y)
Figure 5 Metrological traceability chain of HCT
Hematocrit refers to the percentage of red blood cells in the total blood volume. It reflects the
ratio of red blood cells to plasma. The results of hematocrit determination are expressed as
volume fractions, not involving matrix effects, and the unit is L/L (%).
Technical File

CLSI/NCCLS, etc.) and authoritative institutions (WHO, BCR, IRMM, NIST, etc.), HCT has the
reference measurement procedure recommended by CLSI/NCCLS (standard H7-A2) and ICSH
(recommendations for reference method for the packed cell volume. ICSH Lab Hematol
7,148-170,2001)，and there is no reference material. We choose a method that is easier to op-
erate, saves finished products and is more authoritative as the reference measurement proce-
dure (standard H15-A3). According to the ISO 17511:2020 document clause 5.3, the traceability
chain and calibration hierarchy of HCT is shown in Figure 5. p.3 is the primary measurement
procedures defining the measurand (HCT). m.4 is manufacturer’s working calibrators. Normal
human fresh blood assigned by the reference method (p.3) is selected as the manufacturer’s
working calibrator. p.5 is manufacturer’s standing measurement procedure which calibrated with
the manufacturer's working calibrator. m.5 is end-user IVD MD calibrators. End-user IVD MD
calibrators assigned through a qualified manufacturing process for manufacturer’s standing
measurement procedure. p.6 is End-user IVD MD. The assigned end-user IVD medical device
calibrators can be used for the calibration of end-of-line hematology analyzers. m.6 is human
sample with result.
2.5 Platelet Count (PLT):

procedures:. Counting and FCM

the measurand (PLT): ① WHO/DIL/00.3 Rec-
ommended Methods for the Visual Determination
m.4.Manufacturer’s of White Blood Cell Count and Platelet Count.②
working calibrators ICSH. Platelet Counting by the RBC/Platelet Ra-
(Fresh blood sam- tio Method. Am J Clin Pathol. 2001.115(3):460-4.
ple)

MD calibrators p.6. End-user IVD MD
%rucalb m.6.Human sample

with result
u(y)
Figure 6 Metrological traceability chain of PLT
Technical File
Platelet count refers to the number of platelets contained in a unit volume of blood. The type of
amount is cell concentration and the unit of platelet count is /L. The matrix in platelet count is all
components in fresh blood except platelets. After consulting relevant international scien-
tific/medical organizations (JCTLM, ICSH, CLSI/NCCLS, etc.) and authoritative institutions
(WHO, BCR, IRMM, NIST, etc.), PLT has the reference measurement procedure recommended
by WHO (Recommended Methods for the Visual Determination of White Blood Cell Count and Platelet
Count.) and ICSH (Platelet Counting by the RBC/Platelet Ratio Method. Am J Clin Pathol.
2001.115(3):460-4.)，and there is no reference material. Both of them are selected for method-
ological comparison. According to the ISO 17511:2020 document clause 5.3, the traceability
chain and calibration hierarchy of RBC is shown in Figure 6. p.3 is the primary measurement
procedures defining the measurand (PLT). The principle of these two reference measurement
procedures is counting: 1) According to the reference method recommended by WHO, dilute
fresh blood samples with 1% ammonium oxalate, and manually count platelets under a micro-
scope. 2) According to the reference method recommended by ICSH, the platelets were fluo-
rescently labeled, measured by flow cytometry, and the ratio of red blood cells to platelets was
counted, and then calculated according to the formula to count the number of platelets in fresh
blood. m.4 is manufacturer’s working calibrators. Normal human fresh blood assigned by the
reference method (p.3) is selected as the manufacturer’s working calibrator. p.5 is manufactur-
er’s standing measurement procedure which calibrated with the manufacturer's working cali-
brator. m.5 is end-user IVD MD calibrators. End-user IVD MD calibrators assigned through a
qualified manufacturing process for manufacturer’s standing measurement procedure. p.6 is
End-user IVD MD. The assigned end-user IVD medical device calibrators can be used for the
calibration of end-of-line hematology analyzers. m.6 is human sample with result.
3 Procedure of Value Assignment

3.1 Value Assignment of Working Calibrator and Source of Uncertainty
The assignment of the working calibrator is in the reference measurement laboratory of Shen-
zhen Dymind Biotechnology Co., Ltd., which was recognized by the China National Accreditation
Service for conformity assessment (CNAS) on October 16, 2019. Take 2 fresh blood samples
from different persons. Blood donors are required to be healthy adults (except pregnant women),
who have no disease, infection, anemia, etc. during blood donation. Every year, blood donors
are organized to go to regular hospitals for physical examination, and the results of blood routine
and basic biochemical tests are required to be normal. Apply the reference measurement pro-
cedures corresponding to the five parameters of WBC, RBC, HGB, PLT, HCT, and perform ex-
perimental tests respectively according to the corresponding measurement reference method’s
operation manual to assign the values of the blood samples preliminarily. One of the blood
sample with assigned values is used as the working calibrator. The principle of each reference
method is presented in a) ~ e), and its uncertainty is provided in the "Reference Measurement
Report"
a WBC count: The electronic cytometer is used to measure the WBC count by linear regres-
sion method. The model of particle analyzer is Multisizer 4e and the equipment control
number is BE13120
b RBC count: The electronic cytometer is used to measure the RBC count by linear regres-
sion method. The model of particle analyzer is Multisizer 4e and the equipment control
number is BE13120
c Hemoglobin measurement (HGB): The cyanmethemoglobin (CN-Hb) measurement method
Technical File
is used to measure the concentration of hemoglobin in the blood sample by a spectropho-

tometer. The model of UV-Vis spectrophotometer is GENESYS 180 and the equipment
control number is 9A5W358001.
d Hematocrit measurement (HCT): The micro-hematocrit centrifuge is used to measure and
calculate the hematocrit. The model of microscope is ECLIPSE E200MV and the equipment
control number is 20170620033.
e PLT count: Platelet count was measured by phase contrast microscope. The model of mi-
croscope is ECLIPSE E100 and the equipment control number is 853602.
Table 1 Assigned values and uncertainty of reference methods
Sample No. Reference value WBC RBC HGB HCT MCV PLT
Assigned value 1 of
7.40 5.65 155 48.5 85.8 234
working calibrator
2022122701
Relative expanded un-
2.7 1.9 1.9 0.6 1.9 5.8
certainty/%
Assigned value 2 of
6.49 4.59 142 42.1 91.7 215
working calibrator
2022122702
Relative expanded un-
2.7 1.9 1.9 0.6 1.9 5.8
certainty/%
3.2 Value Assignment of Product Calibrator and Source of Uncertainty

3.2.1 Validation of Reference Measurement Procedure
Use Dymind hematology analyzer DF50 that is calibrated with working calibrator (Manufacturer's
standing measurement procedure), confirmed that the instrument functions normally and its
performance state meets the requirements of the operating instructions of the hematology ana-
lyzer and one of the blood sample with assigned values for calibration confirmation. Test for 11
times in a row, discard the first test result to prevent contamination, and calculate the average of
measurements. And use the assigned values of the reference method as the target values to
calculate the bias between the average of measurements and the target values. If WBC≤1.5%,
RBC≤1.0%, HGB≤1.0%, MCV≤1.0%, PLT≤3.0%, the reference measurement procedure is val-
idated.
Table 2 Calibration verification of blood cell analyzer
Parameter WBC RBC HGB MCV PLT
Reference 6.49 4.59 142 91.7 215

value
Technical File
Average of 6.45 4.55 141 91.9 220

measurements
Bias B (%) 0.62 0.87 0.70 0.22 2.33
Standard B ≤1.5 ≤1.0 ≤1.0 ≤1.0 ≤3.0

(%)
Judgment Qualified Qualified Qualified Qualified Qualified
3.2.2 Method of Value Assignment

Use the qualified calibration detection system in 3.2.1 to evaluate the calibrator according to the
Evaluation Process of Calibrators for Blood Cell Analyzers. Calculate the CV of uniformity in the
bottle and the F-test value. If WBC ≤ 4%, RBC≤2.0%, HGB≤1.5%, MCV≤3.0%, PLT≤5.0% and F≤F
（0.05，v1,v2）,the measured mean value is the initial fixed value of the calibrator.
Table 3 Value Assignment Data of Hematology Calibrator
Reference value 9.33 4.70 136 84.5 256
CV/% 1.85 1.01 0.82 1.08 2.31
Standard CV/% ≤4.0 ≤2.0 ≤1.5 ≤3.0 ≤5.0
F 1.106 2.269 1.900 1.497 2.108
𝐹𝐹(0.05,𝑉𝑉1 ,𝑉𝑉2) 2.393 2.393 2.393 2.393 2.393
3.2.3Commutability of Calibrator
Commutability is one of the important characteristics of the calibrator. Whether it has commuta-
bility means whether it has a matrix effect, that is, the performance of the calibrator should be
consistent with the performance of the sample to be tested. Use the above-mentioned hema-
tology analyzer for testing, and use the product calibrator whose value is initially assigned as the
standard for calibration. Select 20 fresh venous blood samples anticoagulated with EDTAK2.
When selecting the sample, it should be noted that its values should be distributed as evenly as
Technical File
possible within the full measurement range of the hematology analyzer. Test each venous blood
sample within 4 hours after collection respectively with reference method (working calibrator)
and the hematology analyzer that is calibrated with the product calibrator. Each sample was
tested 3 times on the blood cell analyzer, and the average value was taken. Three calibrators
were randomly selected and tested by the same method, and the calibrators were randomly in-
terspersed in the new blood samples. Record the corresponding results, perform the correlation
analysis, calculate the correlation coefficient, slope, intercept, and establish linear regression
equation to prove the commutability of manufacturer product calibrator. The result of the hema-
tology analyzer calibrated by product calibrator should be correlated with the result of the he-
matology analyzer calibrated by working calibrator, and the correlation coefficient r≥0.975 (or
equivalent to r2≥0.95), then the value assigned by the product calibrator can be regarded as re-
liable.
Procedure 1: Manufacturer's standing measurement procedure
Procedure 2: End-user IVD MD
Table 4 Correlation Data of Calibration Results
Pro- Pro- Pro- Pro- Pro- Pro- Pro- Pro- Pro- Pro-
Sam-
cedure cedure cedure cedure cedure cedure cedure cedure cedure cedure
ple No.
1 2 1 2 1 2 1 2 1 2
WBC WBC RBC RBC HGB HGB MCV MCV PLT PLT
1 9.4 9.62 3.20 3.13 70 71 92.7 93.3 249 250
2 7.23 7.11 2.76 2.83 109 110 97.1 99.3 217 219
3 5.8 5.68 4.27 4.32 159 161 90.5 89.4 205 206
4 5.76 5.84 2.03 2.05 144 144 104.6 104.8 226 225
5 10.37 10.37 4.72 4.82 127 127 96.5 96.5 232 226
cali-
bra- 9.33 9.35 4.77 4.75 135 137 84.7 84.7 252 257
tor1
6 12 11.92 2.62 2.67 136 136 84.4 84.2 265 260
7 11.83 11.63 4.64 4.49 175 175 81.7 82.8 388 402
8 6.91 6.91 5.11 5.13 141 141 75.7 74.4 258 262
9 5.96 5.81 4.59 4.54 142 142 68.7 68.5 257 262
10 7.25 7.41 4.62 4.45 132 130 70.9 70.3 296 297
Technical File
Pro- Pro- Pro- Pro- Pro- Pro- Pro- Pro- Pro- Pro-
Sam-
cedure cedure cedure cedure cedure cedure cedure cedure cedure cedure
ple No.
1 2 1 2 1 2 1 2 1 2
WBC WBC RBC RBC HGB HGB MCV MCV PLT PLT
11 3.29 3.23 5.21 5.16 146 146 95.4 95.2 97 94
12 5.7 5.63 3.84 3.86 133 133 91.2 92.3 239 235
13 4.86 4.83 5.32 5.26 164 164 98 98.0 375 375
cali-
bra- 9.36 9.44 4.78 4.72 136 137 84.4 84.2 255 254
tor2
14 4.56 4.39 4.82 4.79 128 128 68.5 69.3 204 211
15 2.05 2.03 3.32 3.29 133 133 93.6 94.7 169 165
16 2.80 2.84 2.74 2.55 154 154 88 88.2 174 173
17 7.2 7.07 4.07 4.04 136 136 103.5 102.9 198 199
18 5.13 5.14 5.07 5.10 122 122 104 103.8 428 445
cali-
bra- 9.37 9.35 4.69 4.70 136 137 84.1 84.1 253 251
tor3
19 5.4 5.33 4.18 4.20 149 148 112.9 110.6 249 250
20 5.46 5.48 4.19 4.17 122 123 113.5 113.8 217 219
y=ax+ y = 0.9979x - y = 0.9943x - y = 0.9894x + y = 0.9928x + y = 1.0464x -

b 0.019 0.0003 1.5416 0.7145 9.5943
r2 0.9984 0.9936 0.9984 0.9946 0.9973
Stand
Correlation coefficient r2≥0.95
ard
Judgm
Qualified Judgment Qualified Judgment Qualified
ent
Technical File
3.3 Uncertainty calculation of calibrator

The relative standard uncertainty introduced by calibration value(𝑢𝑢𝑟𝑟𝑟𝑟𝑟𝑟,𝑐𝑐ℎ𝑎𝑎𝑎𝑎 ) ,the relative standard
uncertainty introduced by uniformity(𝑢𝑢𝑟𝑟𝑟𝑟𝑟𝑟,𝑏𝑏𝑏𝑏 ) and the relative standard uncertainty introduced by
stability(𝑢𝑢𝑟𝑟𝑟𝑟𝑟𝑟,𝑠𝑠 ） are calculated according to According to YY/T 1709 Evaluation of Uncertainty of
Calibrator for In Vitro Diagnostic Reagents, and the synthetic relative standard uncertainty of
calibrator is calculated as follows:
2
𝑢𝑢𝑟𝑟𝑟𝑟𝑟𝑟,𝑐𝑐 = �𝑢𝑢𝑟𝑟𝑟𝑟𝑟𝑟,𝑐𝑐ℎ𝑎𝑎𝑎𝑎 + 𝑢𝑢2𝑟𝑟𝑟𝑟𝑟𝑟,𝑏𝑏𝑏𝑏 + 𝑢𝑢2𝑟𝑟𝑟𝑟𝑟𝑟,𝑠𝑠
When coverage factor k=2, its relative expanded uncertainty Urel=K×urel,c=2×urel,c (K=2 means the
confidence level is 95%)
See table 5:
Table 5 Summary of Product Calibrator Uncertainty
Uncertainty WBC RBC HGB MCV PLT
𝑢𝑢𝑟𝑟𝑟𝑟𝑟𝑟,𝑐𝑐ℎ𝑎𝑎𝑎𝑎 /% 1.37 0.99 0.97 0.97 2.94
𝑢𝑢𝑟𝑟𝑟𝑟𝑟𝑟,𝑏𝑏𝑏𝑏 /% 0.24 0.89 0.51 0.40 1.44
𝑢𝑢𝑟𝑟𝑟𝑟𝑟𝑟,𝑠𝑠 /% 1.01 0.75 0.52 0.59 0.63
urel,c/% 1.72 1.53 1.21 1.20 3.34
Assigned value of
9.33 4.70 134 84.5 256
the calibrator
Relative expanded
uncertainty (K=2) 3.5 3.1 2.5 2.4 6.7
Urel (%)
4 Commutability Evaluation of the Calibrator

Commutability is one of the important characteristics of the calibrator. Whether it has commuta-
bility means whether it has a matrix effect, that is, the performance of the calibrator should be
consistent with the performance of the sample to be tested. For the calibrator, even if the as-
signed value is accurate, if it does not have commutability, it will still result in deviations in the
accuracy of the measurement results. This experiment is carried out according to the method of
evaluating commutability recommended by EP14/A3 Evaluation of matrix effects, approved
guideline (CLSI/NCCLS: 2005).
1. Use the standing measurement procedure calibrated by the working calibrator and the
conventional measurement procedure calibrated by the product calibrator to test a series of
clinical samples and product calibrators at the same time. Take the arithmetic mean of the
concentration values measured by the standing measurement procedure as the X axis and
Technical File
the arithmetic mean of the concentration values measured by the conventional measure-
ment procedure as the Y axis to draw the scatter diagram and perform linear regression
analysis.
2. Draw a regression line and a 95% prediction interval line on the scattergram. Observe the
positions of the results on the scattergram. If they fall within the 95% prediction interval line,
then it indicates that it has the commutability; otherwise, it has no commutability, indicating
that there is a certain difference in matrix effect between the product calibrator and the
clinical sample. The final assigned values of product calibrator need to be adjusted through
methodology.
The evaluation scattergram of the commutability of the product calibrator, the 95% prediction
interval line and the regression line are shown in the figure below.
Technical File
5 Result Analysis and Conclusion

According to ISO 17511:2020, In vitro diagnostic medical devices—Requirements for establish-
ing metrological traceability of values assigned to calibrators, trueness control materials and
human samples. the traceability of hematology calibrator is carried out by international reference
method.
In accordance with the guidance of ISO/IEC Guide 98-3:2008 Uncertainty of measurement - Part
3: Guide to the expression of uncertainty in measurement (GUM: 1995).
According to the commutability evaluation method recommended by EP14/A3 Evaluation of
matrix effects, approved guideline (CLSI/NCCLS: 2005), the product calibrator of hematology
calibrator is evaluated for commutability. The actual measurement results are within the 95%
confidence interval of the expected values. There is no obvious matrix difference from the clini-
cal samples, and the commutability is good.

Hematology Calibrator Traceability Report

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Hematology Calibrator Traceability Report

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Reference Measurement Laboratory of

Shenzhen Dymind Biotechnology Co., Ltd.

Traceability Research Report

Calibrator Model: DM-CAL PLUS

Calibrator Lot No./Exp. NK0123/2023-02-10

Written by: LIU Jia

Reviewed by: LIU Jinying

Approved by: ZHENG Jiantong

Release Date: 2022/12/28

Controlled or Not: ■ Yes □ No

1 Basis of Value Assignment

2 Standard of Value Assignment

p.1.set of fit for purpose primary measurement pro-

p.3. primary measurement procedures defining the

p.5.Manufacturer’s standing measurement procedure

%rucalb m.5.End-user IVD MD

%ru(y)c m.6.Human sample

2.1 White blood cell count (WBC):

p.1. set of fit for purpose primary measurement

p.3. primary measurement procedures defining

m.5. End-user IVD

m.6. Human sample

m.6 is human sample with result.

2.2 Red blood cell count (RBC)

p.1.set of fit for purpose primary measurement

p.3. primary measurement procedures defining

%rucalb m.5.End-user IVD

p.1.set of fit for purpose primary measurement

p.3. primary measurement procedures defining

p.6. End-user IVD MD

Figure 4 Metrological traceability chain of HGB

p.1.set of fit for purpose primary measurement

p.3. primary measurement procedures defining

%rucalb m.5.End-user IVD

%ru(y)c m.6.Human sample

Figure 5 Metrological traceability chain of HCT

After consulting relevant international scientific/medical organizations (JCTLM, ICSH,

Material Measurement procedure

p.1.set of fit for purpose primary measurement

p.3. primary measurement procedures defining

%rucalb m.5.End-user IVD

%rucalb m.6.Human sample

3 Procedure of Value Assignment

is used to measure the concentration of hemoglobin in the blood sample by a spectropho-

3.2 Value Assignment of Product Calibrator and Source of Uncertainty

Table 2 Calibration verification of blood cell analyzer

Parameter WBC RBC HGB MCV PLT

Reference 6.49 4.59 142 91.7 215

Parameter WBC RBC HGB MCV PLT

Average of 6.45 4.55 141 91.9 220

Bias B (%) 0.62 0.87 0.70 0.22 2.33

Standard B ≤1.5 ≤1.0 ≤1.0 ≤1.0 ≤3.0

Judgment Qualified Qualified Qualified Qualified Qualified

3.2.2 Method of Value Assignment

Table 3 Value Assignment Data of Hematology Calibrator

Parameter WBC RBC HGB MCV PLT

Reference value 9.33 4.70 136 84.5 256

CV/% 1.85 1.01 0.82 1.08 2.31

Standard CV/% ≤4.0 ≤2.0 ≤1.5 ≤3.0 ≤5.0

Judgment Qualified Qualified Qualified Qualified Qualified

F 1.106 2.269 1.900 1.497 2.108

𝐹𝐹(0.05,𝑉𝑉1 ,𝑉𝑉2) 2.393 2.393 2.393 2.393 2.393

Judgment Qualified Qualified Qualified Qualified Qualified