ECAT FOUNDATION
External quality Control of diagnostic Assays and Tests
with a focus on Thrombosis and Haemostasis
SURVEY MANUAL 2014
All rights reserved. No part of this manual may be reproduced, stored in a retrieval system, or transmitted in any form or by
means, electronic, mechanical, photocopying, recording or otherwise, without prior permission from the ECAT Foundation
ECAT FOUNDATION
External quality Control of diagnostic Assays and Tests
with a focus on Thrombosis and Haemostasis
CONTENT
1. ECAT OFFICE INFORMATION ....................................................................................................................... 4
2. SUBCONTRACTED ACTIVITIES .................................................................................................................... 4
3. MISSION .......................................................................................................................................................... 4
4. GENERAL INFORMATION .............................................................................................................................. 5
5. COMPLAINTS AND APPEALS ........................................................................................................................ 6
6. USE OF RESULTS .......................................................................................................................................... 6
7. SURVEY PROGRAMME 2014 ........................................................................................................................ 7
8. SURVEY SCHEDULE 2014 ............................................................................................................................. 8
9. SAMPLES ........................................................................................................................................................ 9
10. SURVEY INFORMATION ............................................................................................................................ 10
11. REPORTING RESULTS .............................................................................................................................. 11
11.1 General information ................................................................................................................................ 11
11.2 Instructions for Lupus Anticoagulant testing .......................................................................................... 12
11.3 Instructions for D-Dimer ......................................................................................................................... 12
12. INSTRUCTIONS FOR REPORT FORMS FOR POSTAL SERVICE / FAX ................................................. 13
12.1 General explanation report forms........................................................................................................... 13
12.2 Explanation APC Resistance report form .............................................................................................. 13
12.3 Explanation Lupus Anticoagulant report form ........................................................................................ 14
13. INSTRUCTIONS FOR RESULT SUBMISSION VIA INTERNET ................................................................. 15
13.1 General Instructions ............................................................................................................................... 15
13.2 Special instructions for the Lupus Anticoagulant module ...................................................................... 17
13.3 Special instructions for the von Willebrand Factor module .................................................................... 19
14. SUBMISSION RESULTS MOLECULAR BIOLOGY SURVEYS .................................................................. 20
15. STATISTICAL EVALUATION....................................................................................................................... 21
15.1 Statistical evaluation of survey results ................................................................................................... 21
15.2 Explanation of Algorithm A ..................................................................................................................... 22
16. INSTRUCTIONS TO DOWNLOAD SURVEY REPORTS............................................................................ 23
17. SURVEY REPORT....................................................................................................................................... 25
17.1 General explanation of the survey report ............................................................................................... 25
ECAT SURVEY MANUAL 2014
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Page 3 of 27
ECAT FOUNDATION
External quality Control of diagnostic Assays and Tests
with a focus on Thrombosis and Haemostasis
1. ECAT OFFICE INFORMATION
Postal address
ECAT FOUNDATION
P.O. BOX 107
2250 AC Voorschoten
The Netherlands
Visiting address
ECAT Foundation
Dobbeweg 1
2254 AG Voorschoten
The Netherlands
Contact details (general)
T. +31 71 3030910
F. +31 71 3030919
E. [email protected]
W. www.ecat.nl
Contact details (financial department)
T. +31 71 3030911
F. +31 71 3030919
E. [email protected]
Registration number with the Chamber of Commerce (KvK) Gouda : 41174102
VAT number: NL802836872B01
Bank office:
ING
Address:
P.O. Box 94780
1090 GT Amsterdam
The Netherlands
Account no.:
6930471
IBAN no.:
NL38 INGB 0006 9304 71
BIC code:
INGBNL2A
Staff:
Name
Dr. P. Meijer
Mrs. A. Veninga
Mrs. M. Van der Voorn
Mrs. G. Zandbergen
Position
Director
Scheme Manager / Quality Manager
Office Manager / Deputy Scheme Manager
Financial Officer / Survey Assistant
2. SUBCONTRACTED ACTIVITIES
ICT-related issues
Laboratory testing
Development and maintenance web-based
data submission, back-up services, web and
database hosting.
Web design and maintenance.
R. Mendes Holding BV –
Rotterdam
Testing for homogeneity and stability.
Erasmus Medical Center –
Rotterdam
Sample preparation
Goedknegt Design - Rotterdam
Affinity Biologicals – Canada
Hyphen Biomed – France
Technoclone – Austria
MCA – The Netherlands
Klinikum Augsburg - Germany
3. MISSION
The primary aim of the ECAT Foundation is to contribute to quality assessment and improvement of clinical
laboratories operating within the field of thrombosis and haemostasis with respect to the diagnosis and
treatment of patients.
ECAT SURVEY MANUAL 2014
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Page 4 of 27
ECAT FOUNDATION
External quality Control of diagnostic Assays and Tests
with a focus on Thrombosis and Haemostasis
4. GENERAL INFORMATION
The ECAT Foundation has been operating External Quality Assessment Programmes (EQAP) in the field of
thrombosis and haemostasis since 1992. The ECAT is an independent and impartial organization.
The ECAT is based in The Netherlands but provides EQAP for assays and tests in the field of thrombosis and
haemostasis on an international scale. The programme is open for every laboratory providing services in the
mentioned discipline.
Board:
The board of the ECAT Foundation serves as the Scientific Advisory Board. Members of the board are well
experienced in the field of thrombosis and haemostasis.
The board consists of the following persons:
Name
Prof. Dr. C. Kluft
Dr. M.P.M. de Maat
Ir. D.C. van Cuilenburg
Dr. F. Haas
Dr. R. Niessen
Dr. O. Paauwe
Dr. H.W. Verbruggen
Specialism
Biochemist
Biochemist/epidemiologists/clinical chemist
Accountant
Clinical chemist
Clinical chemist
Director Thrombosis Foundation Netherlands
Biochemist
Board function
Chairman
Secretary/Deputy chairman
Treasurer
Report committee:
The reports conducted from the survey results are reviewed by a Report Committee. This committee exists of:
Dr. J. Ruinemans – Koerts (clinical chemist)
Dr. R. Niessen (clinical chemist)
Dr. P. Meijer (Director ECAT)
Accreditation:
The External Quality Assessment Programme of the ECAT Foundation is accredited according
to ISO/IEC 17043:2010 by the Dutch Council for Accreditation (RvA).
More details, including the scope of accreditation, can be found at our website.
The following modules of ECATs’ EQAP are part of the accreditation scope:















Thrombophilia module
Protein C Pathway Test
Lupus Anticoagulant / Antiphospholipid Antibodies
D-Dimer
Coagulation Factor module I
Coagulation Factor module II
Von Willebrand Factor module
Factor VIII inhibitor
Thrombin Generation Test
Factor XIII
Fibrinolysis parameters I
Fibrinolysis parameters II
Screen Module
Monitoring for Anticoagulation Drugs
Homocysteine
ECAT SURVEY MANUAL 2014
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Page 5 of 27
ECAT FOUNDATION
External quality Control of diagnostic Assays and Tests
with a focus on Thrombosis and Haemostasis
5. COMPLAINTS AND APPEALS
Any complaint regarding the delivery of samples, result submission and reports should be directed in written to
the ECAT office (for contact details: see ECAT office information at page 4)
Any complaint regarding a survey report should be sent to the ECAT within 6 weeks
after the issue date of the report.
Complaints received after this period are no longer considered.
Any appeal regarding the activities of the ECAT Foundation should be directed in written to the board of ECAT.
Contact details of the board:
Board ECAT Foundation
attn. Secretary
P.O. Box 107
2250 AC Voorschoten
The Netherlands
6. USE OF RESULTS
Results are treated confidential. Data represented in the reports are not traceable to a specific participant,
except the own results presented in `your results` in the participants´ own report.
Survey results may be used for scientific purposes. In this case anonymous use of results will be guaranteed.
Summaries of survey results may be provided to our suppliers of control samples for both internal use as well as
use in data sheets of control samples. This information will never include individual survey results. Individual
survey results will never be provided to third parties without permission of the participant.
ECAT SURVEY MANUAL 2014
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Page 6 of 27
ECAT FOUNDATION
External quality Control of diagnostic Assays and Tests
with a focus on Thrombosis and Haemostasis
7. SURVEY PROGRAMME 2014
The regular surveys of the ECAT External Quality Assessment Programme include the following modules:
Survey composition
Module
2014-1
2014-2
2014-3
2014-4
Screen module (APTT, PT/INR, Fbg) *
√
√
√
√
Thrombophilia module
Antithrombin (activity and antigen), Protein C (activity [chromogenic and
clotting] and antigen), Protein S activity, Protein S antigen (total and free),
APC Resistance
√
√
√
√
Protein C Pathway Test
√
√
√
√
Lupus Anticoagulant / Antiphospholipid Antibodies
√
√
√
√
D-Dimer
√
√
√
√
Coagulation Factor module I (Factor VIII, IX, XI and XII)
√
√
√
√
Coagulation Factor module II (Factor II, V, VII and X)
√
√
√
√
Von Willebrand Factor module
(antigen, activity, collagen binding, multimers, Factor VIII)
√
√
√
√
ADAMTS-13 (activity and antigen)
√
√
√
√
ADAMTS-13 (inhibitor)
√
√
√
√
Factor XIII
√
√
√
√
Fibrinolysis parameters I (Plasminogen, Antiplasmin)
√
√
√
√
Fibrinolysis parameters II (t-PA, PAI-1)
√
√
√
√
Unfractionated Heparin (anti-Xa)
√
√
√
√
Low-Molecular Weight Heparin (anti-Xa)
√
√
√
√
Homocysteine
√
√
√
√
Factor VIII inhibitor
-
√
-
√
Factor IX inhibitor
√
-
√
-
Thrombin Generation Test
-
√
-
√
HIT – Immunological assays
-
-
√
-
Orgaran (quantitative)
-
√
-
√
Fondaparinux (quantitative)
-
√
-
√
Rivaroxaban (quantitative)
-
√
-
√
Apixaban (quantitative)
-
√
-
√
Argatroban (quantitative)
-
√
-
√
Dabigatran (quantitative)
-
√
-
√
* Note: The screen module is only available in a selected number of countries
Electronic post-analytical platelet function survey:
In co-operation with the NASCOLA in the United States twice a year a post-analytical survey for platelet function
testing is provided. NASCOLA is responsible for the content and evaluation of these surveys. Participants of
these surveys are informed about the details of these surveys by e-mail.
ECAT SURVEY MANUAL 2014
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ECAT FOUNDATION
External quality Control of diagnostic Assays and Tests
with a focus on Thrombosis and Haemostasis
Electronic pre- and post-analytical surveys in haemostasis and case studies in bleeding disorders:
In co-operation with INSTAND in Germany these pre- and post-analytical electronic survey in haemostasis and
case studies in bleeding disorders are provided. Participants of these surveys are informed about the details of
these surveys by e-mail.
Molecular Biology:
In co-operation with the DGKL in Germany an EQA programme for Molecular Diagnostic Testing is provided.
DGKL is responsible for the content, sample distribution and evaluation of these surveys. Samples are
distributed according to the survey schedule shown in paragraph 8.
8. SURVEY SCHEDULE 2014
Every year the ECAT Foundation prepares a Survey Schedule. In this schedule information is given about:
 Survey:
code number of the survey
 Dispatch date:
date that we send the samples to all participants
 Closing date:
date that result submission closes (fax/postal service and website)
 Closing date D-Dimer: date that D-Dimer result submission closes (shorter turn-over time)
This survey schedule includes also the dispatch and closing dates of the molecular biology modules provided by
the DGKL.
All participants receive a copy of this schedule together with information about the annual subscription, so they
know in advance when to expect the samples.
Survey Schedule 2014
PLASMA TESTS MAIN PROGRAMME
(ECAT Foundation, The Netherlands)
Survey
Dispatch Date
Closing Date
Closing Date
Screen / D-Dimer
2014-1
4 March
2 April
19 March
2014-2
20 May
25 June
4 June
2014-3
26 August
1 October
10 September
2014-4
11 November
17 December
26 November
MOLECULAR BIOLOGY, DNA SEQUENCING AND DNA ISOLATION
(via DGKL, Germany)
Survey
Type
FV1/14
Molecular Biology A-I, first survey
FV2/14
Molecular Biology A-I, second survey
SQ1/14
DNA Sequencing, first survey
SQ2/14
DNA Sequencing, second survey
DI1/14
DNA Isolation, first survey
DI2/14
DNA Isolation, second survey
ECAT SURVEY MANUAL 2014
Dispatch Date
Closing Date
19 March
12 April
24 September
18 October
29 January
22 February
3 September
27 September
12 March
5 April
3 September
27 September
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Page 8 of 27
ECAT FOUNDATION
External quality Control of diagnostic Assays and Tests
with a focus on Thrombosis and Haemostasis
9. SAMPLES
General information
The samples used in the surveys are plasma and serum based. Part of the samples are from commercial
source. For abnormal samples real patient plasma is used when appropriate. In the surveys only non-expired
samples are used. Samples are provided as lyophilized material. Instructions for reconstitution are given in the
survey instruction leaflet.
In the survey reports information is given on the homogeneity and stability of the samples used.
The table below shows for each of the modules the type of sample material used and the intended measuring
range included in the surveys.
Module
Screen module:
APTT
PT/INR
Fibrinogen
Thrombophilia module:
Antithrombin (activity and antigen), Protein C (activity
[chromogenic and clotting] and antigen), Protein S
activity, Protein S antigen (total and free), APC
Resistance
Protein C Pathway Test
Lupus Anticoagulant / Antiphospholipid
Antibodies
D-Dimer
Coagulation Factor module I
(Factor VIII, IX, XI and XII)
Coagulation Factor module II
(Factor II, V, VII and X)
Von Willebrand Factor module
(antigen, activity, collagen binding, multimers, Factor
VIII)
ADAMTS-13 (activity and antigen)
Number
of
surveys
per year
Number of
different
samples
per survey
Sample
material
(lyophilized)
Measuring range
(approx.)
4
2
plasma
normal – prolonged
normal – prolonged
100 – 400 mg/dL
4
2
plasma
20 – 120%
normal / FV Leiden
4
2
plasma
normal / abnormal
4
1
plasma
negative - positive
4
2
plasma
normal - elevated
4
2
plasma
0 – 200%
4
2
plasma
0 – 200%
4
1
plasma
0 – 125%
4
2
plasma
0 – 125%
ADAMTS-13 (inhibitor)
4
2
plasma
0 – 10 BU/mL
Factor XIII
Fibrinolysis parameters I
(Plasminogen, Antiplasmin)
Fibrinolysis parameters II
(t-PA, PAI-1)
Unfractionated Heparin (anti-Xa)
4
2
plasma
0 – 120%
4
2
plasma
0 – 120%
4
2
plasma
0 – 50 ng/mL
4
2
plasma
0 – 1.25 IU/mL
Low-Molecular Weight Heparin (anti-Xa)
4
2
plasma
0 – 1.25 IU/mL
Homocysteine
4
2
serum
0 – 100 µmol/L
Factor IX inhibitor
2
2
plasma
0 – 5 BU/mL
Factor VIII inhibitor
2
2
plasma
0 – 5 BU/mL
Thrombin Generation Test
2
3
plasma
normal / abnormal
HIT – Immunological assays
1
2
plasma/serum
negative / positive
Orgaran (quantitative)
2
2
plasma
therapeutic range
Fondaparinux (quantitative)
2
2
plasma
therapeutic range
Rivaroxaban (quantitative)
2
2
plasma
therapeutic range
Apixaban (quantitative)
2
2
plasma
therapeutic range
Argatroban (quantitative)
2
2
plasma
therapeutic range
Dabigatran (quantitative)
2
2
plasma
therapeutic range
ECAT SURVEY MANUAL 2014
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Page 9 of 27
ECAT FOUNDATION
External quality Control of diagnostic Assays and Tests
with a focus on Thrombosis and Haemostasis
Distribution of samples
Samples are sent to participants according to the survey schedule and survey composition. The frequency is
indicated in paragraph 7.
The samples are packed in plastic bubble bags and carton boxes to prevent damage during transport. After
receipt the samples should be stored at 2-8 °C until use.
10. SURVEY INFORMATION
Survey instructions
Each sample shipment is accompanied with specific survey instructions. Read these instructions carefully. This
instruction leaflet gives information which sample should be used for which analyte, volume for reconstitution,
closing date for returning results, etc.
Survey instructions are also available in the participant area on our website.
Sample identification
Samples are properly identified with labels containing the following information:
Sample number, volume for reconstitution, survey number and module identification.
An example of a label is given below.
Sample : 13.03
ECAT
Volume
Survey
Module
: 0.75 mL
: 2013-1
: Thrombophilia
Instructions for reconstitution
For proper reconstitution of the plasmas, please follow exactly the instructions below:

The vial must reach room temperature before adding distilled water.

Dissolve the content of each vial in sterile, distilled, room temperature water. For the exact volume of water
to be used: see survey instruction leaflet.

Leave the vial for 5 minutes.

Swirl the vial gently to mix and leave for a further 15 minutes for complete reconstitution.

Before use mix the vial again gently.
Reconstituted plasma should preferably be used within 1 hour after reconstitution. Plasma should be stored at
room temperature after reconstitution. For immunological methods the reconstituted plasma can be stored for 1
months at -20°C.
Use of Latex Immuno Assays
For the application of ECAT plasmas into Latex-Immuno-Assays test we advise you to centrifuge the
reconstituted plasma for at least 10 minutes at 10.000 x g.
Sample application
The ECAT plasmas should be treated as regular patient plasmas and included in the normal daily analytical
process in the laboratory. The regularly used methods should be applied. No special treatment of the samples is
allowed. Results should be reported similar as a result of a patient is reported.
List with codes for methods and equipment
For participants reporting their results by fax or postal service a separate list with codes for methods and
equipment is enclosed. During the year only updates will be provided when necessary.
The complete list is also available in the participant area on our website.
ECAT SURVEY MANUAL 2014
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Page 10 of 27
ECAT FOUNDATION
External quality Control of diagnostic Assays and Tests
with a focus on Thrombosis and Haemostasis
11. REPORTING RESULTS
11.1 General information
Results should be preferably submitted via the results submission facility in the participants’ area of our website.
If you do not have a password to login for this facility, please contact the ECAT office.
For instructions about the use of the result submission facility see paragraph 13.
If you are not able to return results via our website, please use only the provided report forms for returning
results. Report forms are only provided to those participants that have registered for returning results via fax or
postal service. For instructions about the use of the report forms see paragraph 12.
Results returned after the closing date of the survey (see survey schedule) will not be
included in the evaluation.
To standardise the display of results these should be reported for each parameter with the below indicated
number of decimals.
Parameter
No of
decimals
No of
decimals
APTT / PT
1
Plasminogen activity
0
INR
2
Plasminogen antigen (%)
0
Fibrinogen
0
Plasminogen antigen (mg/dL)
1
Antithrombin activity
0
Plasmin Inhibitor
0
Antithrombin antigen (%)
0
PAI-1 antigen
1
Antithrombin antigen (mg/dL)
1
PAI-1 activity
1
Protein C (activity and antigen),
0
t-PA antigen
1
Protein S (activity and antigen)
0
2
APC Resistance (ratio)
2
Protein C Pathway Test (ratio)
2
Anti-Xa (UFH / LMWH)
Anti-Xa (Orgaran / Fondaparinux /
Rivaroxaban, Apixaban)
Argatroban (anti-IIa / dTT)
Lupus anticoagulant (clotting times)
1
Dabigatran (anti-IIa / dTT)
0
Lupus anticoagulant (ratio)
2
Homocysteine
1
D-Dimer (ng/mL)
0
Factor VIII inhibitor (titer)
1
D-Dimer (mg/L)
Coagulation Factor activity
(Factor II, V, VII, VIII, IX, X, XI and XII)
Von Willebrand Factor module
(antigen, activity, collagen binding, FVIII)
ADAMTS13 (activity / antigen)
2
Factor IX inhibitor (titer)
1
0
Thrombin Generation lag time (min)
1
0
Thrombin Generation time to peak (min)
1
0
Thrombin Generation peak thrombin (nM)
0
ADAMTS13 (inhibitor)
1
Thrombin Generation Velocity Index
1
Factor XIII activity
0
Thrombin Generation ETP/AUC
0
Factor XIII antigen (%)
0
HIT (OD)
3
Factor XIII antigen (mg/dL)
1
HIT (U/mL)
1
0
Parameter
HIT (%)
ECAT SURVEY MANUAL 2014
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2
2
Page 11 of 27
ECAT FOUNDATION
External quality Control of diagnostic Assays and Tests
with a focus on Thrombosis and Haemostasis
11.2 Instructions for Lupus Anticoagulant testing
According to the guidelines of the Standardization and Subcommittee (SSC) of the International Society of
Thrombosis and Haemostasis (ISTH) for Lupus Anticoagulant (LA) a laboratory should perform:
 At least 2 different screening tests based on different methodology (e.g. APTT and DRVVT).
 A mixing study should be performed to exclude whether a prolonged clotting time was related to a
factor deficiency.
 It should be confirmed that the prolonged clotting time is phospholipid-dependent.
Therefore each test panel exists of three different tests, a screening test, a mixing study and a confirmation test.
The ECAT External Quality Assessment Programme provides the ability to report the results for three different
test panels.
These different test panels are not meant to report repeated measurements with the same
reagent.
Additional information for users of the STAGO Staclot LA method:
This test is an APTT-based confirmation test and should be reported as such.
The result obtained with this test is reported as a difference in measured clotting times. It should be reported
with the unit “Delta seconds”.
Normal plasma
In the evaluation of the reported test results the ratio between the ECAT plasma and the laboratories own
normal plasma is calculated. To make the calculation of this ratio possible the measured clotting time for the
normal plasma should be reported.
The report box for normal plasma is not meant to report a reference range for normal plasma.
Ratio screen/confirm
The ratio screen / confirm is frequently used as an evaluation tool to assess the LA positivity of a sample. Here
the ratio of the result of the screening test and the confirmation test should be reported. No other ratios should
be reported in this result box.
11.3 Instructions for D-Dimer

Selection of units:
Please select the appropriate units. Use only the original units of the method
(i.e. D-Dimer units or FEU).
Also select within the unit-group the correct measuring unit: ng/mL or mg/L.
When incorrect units are selected the results will not be included in the survey
evaluation!

Tinaquant method:
Citrate plasma
If the method is calibrated for the use of citrated plasma select in the method list
the Tinaquant (citrate plasma).
Heparin plasma
Because ECAT always provides citrated plasma in their surveys you have to
correct your results before submission in the case you use heparin plasma.
Please multiply the measured result with the correction factor 1.19.
If the method is calibrated for the use of heparin plasma select in the method list
the Tinaquant (heparin plasma).
ECAT SURVEY MANUAL 2014
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ECAT FOUNDATION
External quality Control of diagnostic Assays and Tests
with a focus on Thrombosis and Haemostasis
12. INSTRUCTIONS FOR REPORT FORMS FOR POSTAL SERVICE / FAX
12.1 General explanation report forms
12.2 Explanation APC Resistance report form
EXAMPLE APC Resistance
Without Factor V deficient plasma
Type of assay
APC Resistance Ratio
for ECAT plasma
Enter the code for the
method
Specify from list
Clotting
Method
Chromogenic
Equipment
Indicate which assay
type is used
APC Resistance Ratio
for Laboratories
Reference Plasma
Calculated Normalised
Ratio = Ratio ECAT
plasma / Ratio
Reference Plasma
Result
Ratio ECAT Plasma
Ratio Laboratories
Reference Plasma
Normalised Ratio
Enter code for the
equipment
Classification
Normal
If abnormal:
Borderline
Abnormal
Heterozygous FVL
Homozygous FVL
Non-conclusive
Indicate the clinical
classification of the
sample.
If abnormal, also
indicate the possible
abnormality
ECAT SURVEY MANUAL 2014
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Page 13 of 27
ECAT FOUNDATION
External quality Control of diagnostic Assays and Tests
with a focus on Thrombosis and Haemostasis
12.3 Explanation Lupus Anticoagulant report form
EXPLANATION LUPUS ANTICOAGULANT REPORT FORMS
TEST PANEL 3
Indicate on which type of
assay your laboratory
method is based.
Enter the code for the
method.
SCREENING TEST 3
Type of assay
APTT
X DRVVT
KCT
dAPTT
dPT (TTI)
other
MIXING STUDY 3
Type of assay
APTT
DRVVT
KCT
dAPTT
dPT (TTI)
other
Specify from list
Method
Equipment
Result
001
21
Indicate the
appropriate unit
Unit
X
115
Please, indicate!
Normal
Seconds
X Prolonged
Ratio
Borderline
Enter the code for the
equipment used
(manual application is
code 0)
Normal plasma
Enter result for ECAT
plasma
35
Different parts of test
panel
Indicate the
classification of the
ECAT sample
Enter result for
laboratories own
normal plasma
Ratio ECAT plasma/Normal Plasma
1+1
X
1+2
1+4
1+5
Indicate for the mixing study the
ratio ECAT plasma / Normal
plasma used
CONFIRMATION TEST 3
Type of assay
APTT
PT
DRVVT
KCT
PNP
dAPTT
dPT
Ratio LA screen / LA cofirm
Specify from list
Method
Equipment
Result
Unit
Seconds
Ratio
Delta Sec.
53.5
Please, indicate!
LA positive
LA negative
Borderline
Normal plasma
Enter the ratio
between the result for
the screeing test and
the confirm test
Indicate if the ratio
screen/confirm is a standard
ratio or a normalised ratio
Ratio
2.15
Please, indicate!
X
Standard ratio
Normalised ratio
Comments
FINAL CONCLUSION
X
Negative
Borderline
Probably positive
Positive
Clearly positive
Additional comments
Indicate the final
interpretation of the
ECAT sample
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ECAT FOUNDATION
External quality Control of diagnostic Assays and Tests
with a focus on Thrombosis and Haemostasis
13. INSTRUCTIONS FOR RESULT SUBMISSION VIA INTERNET
FOR ANY PROBLEM WITH RESULT SUBMISSION PLEASE CONTACT THE ECAT
OFFICE: [email protected]
IMPORTANT INFORMATION

For decimals only use . (dots) and no comma’s.

Do not add text to the result fields.
13.1 General Instructions
1. Homepage


Start ECAT website: www.ecat.nl
Select at ECAT homepage the option:
Login
2. Login



Before you can enter the participant area you have to login.
Enter your
Username (=Labcode)
and your
Password
(provided by ECAT)
Select:
Log me in
3. Participant Area


You are now in the Participant Area of the ECAT website.
Select in the menu: Result submission
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ECAT FOUNDATION
External quality Control of diagnostic Assays and Tests
with a focus on Thrombosis and Haemostasis
4. Result submission


You are now at the page where you can go to the Result
Submission facility.
Select in Submit Results
(You are now directed to the result submission facility. In some
cases it may happen that there is a communication failure with
the data base for result submission. In such an occasion an
extra login is necessary. This will be indicated at the screen)
At this page also important instructions can be given from the
ECAT office and/or webmaster regarding results submission
issues.
5. Selection of module


On your screen you will see an overview of all modules you
have subscribed for.
Select the module for which you will enter results,
e.g. Thrombophilia (click on “Thrombophilia”).
6. Selection of Analyte


On your screen you will see an overview of all the tests
belonging to the selected module.
Select under Assay the analyte for which you will enter
results (click on the name of the analyte),
e.g. Antithrombin (click on “Antithrombin”).
When a resubmission is necessary in the case of an observed
error, please select again the analyte in the Assay list.
The last submitted result will be used for statistical analysis.
7. Enter results






Enter results:
Complete the fields for assay, method, equipment, result, unit
and interpretation (use blue arrows to select options).
All the mentioned fields are required! Results can not
be submitted if one of the fields is not completed.
If you have any comments to add, the text box for notes can be
used. Do not add comments to the result fields.
.
For decimals use only (dots) and no comma’s.
If no exact quantitative test results can be given (e.g. the test
results is above the upper limit of the calibration curve –
e.g. > 120% -) enter in the number in the result field and select
in the field < or > the appropriate indicator.
Select:
Print (print form for your own files).
Select:
Submit (to send results to the ECAT database).
ECAT SURVEY MANUAL 2014
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ECAT FOUNDATION
External quality Control of diagnostic Assays and Tests
with a focus on Thrombosis and Haemostasis
8. Submission confirmation


After correct completion of the result sheet and submission of
the results a confirmation of a successful result submission will
be given.
Here the option can be selected to return to the list of assays of
the module selected before or return to the list of modules
belonging to your subscription profile.
9. View function

The view function is only meant to review your last submitted
results. When you have submitted results the date of
submission, the sample codes and your participant number will
appear in the header. So here you can check your latest
submission yourself.
13.2 Special instructions for the Lupus Anticoagulant module
The Lupus Anticoagulant Module consist of several panels. First a selection should be made for which parts of a
panel test results will be submitted. Activate the panel by selecting the panel button. After data entry of the first
panel you can return to the main screen of the Lupus Anticoagulant module where you can continue with the
next panel. See for details instructions below.
1.
Data entry Lupus Anticoagulant module:

Select the appropriate parts of panel 1:
In this example the screening and confirmation tests are
selected.

Select: Panel 1
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ECAT FOUNDATION
External quality Control of diagnostic Assays and Tests
with a focus on Thrombosis and Haemostasis
2.
Complete the results sheet for panel 1.
(Same procedure as described in the general instructions)
.


Use for decimals only (dots) and no comma’s.
Do not add comments to the result fields.

Select:

Select:
Print
(print form for your own files).
Submit
(to send results to the ECAT database).
3.
If you would like to submit results for more Lupus Anticoagulant
panels or for Phospholipid Antibodies, please select Assays.
You will return to the main Lupus Anticoagulant screen where you
can select the next panel. Repeat step 1 – 3 when appropriate.
4.
After submission of all your results, please select
Final conclusion.
5.
Select the appropriate conclusion and submit.
After submission you can return to the list of modules.
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ECAT FOUNDATION
External quality Control of diagnostic Assays and Tests
with a focus on Thrombosis and Haemostasis
13.3 Special instructions for the von Willebrand Factor module
The Von Willebrand Factor Module consist of several test. Here first a selection should be made for which
analytes test results will be submitted. After activation of the selection only the selected test will be shown for
data entry. See instructions below.
1.
Data entry VWF module:

Select the appropriate tests:
In this example VWF antigen and ristocetin cofactor activity
are selected.

Select: Activate Selection
2.
Complete the result sheet as described above.
(Same procedure as described in the general instructions)
.


Use for decimals only (dots) and no comma’s.
Do not add comments to the result fields.

Select:

Select:
Print
(print form for your own files).
Submit
(to send results to the ECAT database).
3.

Before you return to the list of modules do not forget to
select and submit your VWF interpretation.
ECAT SURVEY MANUAL 2014
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ECAT FOUNDATION
External quality Control of diagnostic Assays and Tests
with a focus on Thrombosis and Haemostasis
14. SUBMISSION RESULTS MOLECULAR BIOLOGY SURVEYS
These surveys are provided by the DGKL-RfB in Bonn Germany.
Results for these modules should be directly submitted to the DGKL-RfB.
Codes for modules:
FV = Molecular Biology
DI = DNA isolation
SQ = DNA sequencing
There are two possibilities for result submission.
1) Via the report forms included with the samples and follow the instructions given by DGKL-RfB.
2) Via the website of DGKL-RfB (https://www.dgkl-rfb.de/cgi/switchLang?lang=en)
Homepage DGKL-RfB:
At the right top corner of the homepage you can login.
A specific participant number and password are needed.
Do not use your ECAT login details here!
To obtain this specific participant number and password
please select the option: “Don’t have an account yet.
Please register”.
Follow the instructions given on the registration page (see
below). The DGKL-RfB will provide you with the
participant number and password by e-mail.
Important note!
In the particicpants area of the DGKL-RfB website you can also order surveys. Don’t use this option. Only
register via the ECAT office. This to avoid double registrations.
Result submission:
First log in at the homepage of the DGKL-RfB website.
Make the appropriate selection for submission of your results.
ECAT SURVEY MANUAL 2014
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ECAT FOUNDATION
External quality Control of diagnostic Assays and Tests
with a focus on Thrombosis and Haemostasis
15. STATISTICAL EVALUATION
15.1 Statistical evaluation of survey results
For the external quality assessment programme of the ECAT the robust average of the results reported by all
participants in the survey is used as the assigned value (= consensus value). In accordance with ISO/IEC
standard 17043:2010 and ISO standard 13528:2005 Algorithm A is used as a robust statistical for the
calculation of the consensus value and the standard deviation.
Further explanation on Algorithm A can be found in paragraph 14.2.
The standard procedure for the evaluation of quantitative test results is as follows:
 Results are harmonised to the same unit (% / U/dL).
 The consensus value and standard deviation (SD) using Algorithm A.
 Based on this consensus value and SD the between-laboratory variation is calculated.
Algorithm A is applied on the total group and the level of assay type and method if there are at least 10
participants included in the same group (for the screen assays and homocysteine the minimum number is 5).
The consensus values are represented in the report as assigned value (see explanation of the report in
paragraph 16.1). For groups with less than 10 participants (for the screen assays and homocysteine less than 5
participants) the median is used as target value.
The measurement uncertainty of the consensus values is displayed by:
UAV = 1.25 x s / √p
U = uncertainty
AV = assigned value
s = standard deviation
p = number of participants
This information can be obtained from the report as follows:
cv and p can be obtained from the report.
s can be calculated from the coefficient of variation (CV) and the consensus value by the following formula:
s = (CV/100) x AV
Performance Indicator
As an individual performance indicator the Z-score is calculated.
Z-score
The z-score is calculated as follows:
[(laboratory result) - (mean result of all laboratories)] / (standard deviation of all results)
The Z-score is al so calculated for groups on the level of assay type and method with at least 10 participants.
Acceptance criteria
Each participants should carefully evaluate the Z-scores given in the report.
In accordance with ISO/IEC standard 17043:2010 and ISO standard 13528:2005 the following acceptance
criteria are used:
-2 < Z-score < 2
-3 < Z-score < -2 or 2 < Z-score < 3
Z-score < -3 or Z-score > 3
:
:
:
The result is acceptable
The results is questionable (warning signal)
The result is unacceptable (action signal)
A single action signal or two warning signals in consecutive exercises shall be taken as evidence that a anomaly
has occurred that requires investigation by the laboratory.
ECAT SURVEY MANUAL 2014
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ECAT FOUNDATION
External quality Control of diagnostic Assays and Tests
with a focus on Thrombosis and Haemostasis
15.2 Explanation of Algorithm A
Algorithm A is mainly based on the robust statistical model of Hampel. From the original data set the initial mean
value (x*) and standard deviation (s*) are calculated. As initial value for x* the median is taken and as initial
value for s* the median absolute deviation (MAD) is calculated. The MAD is defined as s* = 1.483 x median |x i –
x*|. In other words, for each value of the data set the absolute deviation of the initial mean value (x*) is
calculated. From this set of deviations the median is taken and multiplied by 1.483. This value is defined as the
initial standard deviation. Then limits for the distribution of the data set are calculated. These limits are defined
as x* ± δ, where δ = 1.5 x s*. If any value of the data set is outside these limits this value is transformed to the
value of the limit. From this new data set a new x* and s* are calculated, where s* is defined as
2
s* = 1.134 x √ ∑(xi”- x*) /(p – 1). [xi” = values from the transformed data set, p = number of results]. With this
new x* and s* new limits (δ) are calculated. These news limits are applied to the original data set, results
outside the limits are transformed to the limit values and new x* and s* are calculated. This procedure is
repeated until the process converges. Convergence may be assumed when there is no change from any one
iteration to the next in the third significant figure of the standard deviation or of the equivalent figure of the mean
value. The final mean value obtained and the standard deviation are used for the calculation of the betweenlaboratory coefficient of variation and the Z-score. The Z-score of an individual result is calculated on the basis
of the value in the original data set. An example of the application of Algorithm A is given in figure 1.
Fig. 1 Example of application of Algorithm
A to a data set with 4 iteration steps.
[Example taken from a presentation by Dr
Koch, University of Stuttgart, Germany,
given at the PT Conference in Sibiu,
Romania 2009.
Background Literature
Daszykowski M. et al (2007). Robust statistics in data analysis. Chemometrics and Intel Lab Systems 85: 203 –
219.
Hampel F. (1974). The influence curve and its role in robust estimation. J. Am. Statist. Assoc. 69: 383 – 393.
Huber P.J. (1964). Robust estimation of a location parameter. Ann. Math. Statist. 35: 73 – 101.
Huber P.J. (1965). A robust version of the probability ratio test. Ann. Math. Statist. 36: 1753 – 1758.
ISO 13528:2005. Statistical methods for use in proficiency testing by interlaboratory comparisons.
ISO 17043:2010. Conformity assessment – General requirements for proficiency testing.
ECAT SURVEY MANUAL 2014
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ECAT FOUNDATION
External quality Control of diagnostic Assays and Tests
with a focus on Thrombosis and Haemostasis
16. INSTRUCTIONS TO DOWNLOAD SURVEY REPORTS
Within the external quality assessment programme of the ECAT Foundation the following survey reports are
distinguished:
Type of survey report
Screen
Main
Modules/Parameters
APTT, PT/INR, Fibrinogen
Thrombophilia, Protein C Pathway, Coagulation
Factor module i and II, Von Willebrand Factor module,
ADAMTS13, Factor VIII inhibitor, Factor IX inhibitor,
Factor XIII, Fibrinolysis module I and II,
Unfractionated Heparin, Low-Molecular Weight
Heparin, Homocysteine Orgaran, Fondaparinux,
Rivaroxaban, Apixaban, Argatroban, Dabigatran,
Lupus Anticoagulant
D-Dimer
HIT
TGT
Lupus Anticoagulant / Antiphospholipid Antibodies
D-Dimer
HIT immunological assays
Thrombin Generation Test
Note: The availability of a particular report and/or modules within the main report depends on the survey composition (see
schedule paragraph 7)
1. Homepage


Start ECAT website: www.ecat.nl
Select at ECAT homepage the option:
Login
2. Login



Before you can enter the participant area you have to login.
Enter your
Username (=Labcode)
and your
Password
(provided by ECAT)
Select:
Log me in
ECAT SURVEY MANUAL 2014
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ECAT FOUNDATION
External quality Control of diagnostic Assays and Tests
with a focus on Thrombosis and Haemostasis
3. Participant Area


You are now in the Participant Area of the ECAT website.
Select in the menu: Survey Reports
4. Download Survey Reports


You are now at the page where you can go to the area where
you can download survey reports.
Select in Download Reports
5. Select and download Survey Report


You are now at the page where you can download survey
reports.
Select from the appropriate survey the report you would like to
download. The last survey is always on the top of the list.
Reports up to two years ago can be downloaded.
In this example the D-Dimer report is selected.
You can now print and/or save the report to your own
computer.
Note: Survey reports are available in PDF-format. To be able
to read the reports it is important that Acrobat Reader is
installed on your computer.
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ECAT FOUNDATION
External quality Control of diagnostic Assays and Tests
with a focus on Thrombosis and Haemostasis
17. SURVEY REPORT
Important general remark:
When results are not correct presented in the survey report caused by a mistake of the
participant (e.g. a wrong result was submitted) no new survey report will be provided.
When results are not correct presented in the survey report caused by a fault of ECAT a
new report will be provided.
From each survey an individual survey report will be provided. Each report includes a front page, general page
and the evaluation of the survey results for those modules a participant has subscribed for. Below a description
of the report is given.
17.1 General explanation of the survey report
Front page
The front page exposes the type of report, survey number and labcode.
General page
This page includes the report issue date, the survey number and the type of report as well as general
information.
Here is also stated that the Survey Manual 2014 should be considered an integral part of the survey report.
Survey report pages
For each analyte a participant has subscribed for in the ECAT programme a report is given.
The report consists of the following parts:
1. The header
2. The graph
3. The table
4. Z-score plot
5. Z–score history plot
The header
The header of each report sheet consist of two parts. At the top of each page the survey number, number of
pages of the report, the date the report is issued and the labcode is indicated. Also the name of the Module and
the analyte is indicated.
Example of a header
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ECAT FOUNDATION
External quality Control of diagnostic Assays and Tests
with a focus on Thrombosis and Haemostasis
In addition, information about the number of responders, the sample used, the units in which the results are
reported, prior use of the sample, stability and homogeneity is given.
Example of general information
When appropriate the clinical classification is given in a separate table.
Example of classification table
The graph
The distribution of the results is represented in a histogram.
Depending of the analyte the results are grouped based on the assay
principle or the method used.
The position of your own result within the distribution is indicated by a black
arrow on top of the corresponding bar in the histogram.
The table
The table given in the report shows the
descriptive statistics for all results and for each
assay principle and method.
The assigned value represents the consensus
value as calculated by Algorithm A.
The between-laboratory variation (CV) and Zscore are only given when at least 10
participants belong to the same group (for the
screen assays and homocysteine this number
is 5).
The group(s) to which your results belong are
highlighted in grey.
ECAT SURVEY MANUAL 2014
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ECAT FOUNDATION
External quality Control of diagnostic Assays and Tests
with a focus on Thrombosis and Haemostasis
Z-score plot
The relationship of the Z-scores of the two different samples are
plotted in a Z-score plot. The Z-score plot only includes methods
with at least 10 participants.
The relationship of both Z-scores gives an indication if the
deviation from the mean value of your particular method is
caused by systematic and/or random errors.
Z-score history
The history of the Z-score for a period of one year is given in a
Z-score history plot.
The dashed lines in the Z-score history plot indicates the level
of -1/1 , -2/2 and -3/3.
For an explanation about the Z-score see paragraph 14.1.
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