PROFICIENCY TESTING REPORT - 岡大バイオバン …biobank.ccsv.okayama-u.ac.jp/.../01/PBMC16R1-L261-Report.pdfservices/pt/proficiency-testing-schemes/). Personalized Advice Service:

PROFICIENCY TESTING REPORT

VIABLE PBMC ISOLATION

Final PBMC16R1 L261 Prepared on: 13 January 2017 Page 2 of 18

Biorepository proficiency testing for the quality control

of biospecimens

Proficiency Testing Provider

IBBL (Integrated Biobank of Luxembourg)

6 Rue Nicolas Ernest Barblé

L-1210 Luxembourg

LUXEMBOURG

Coordinator Fay BETSOU, PhD HDR, IBBL CBS

Contact us Tel: +352 26970 - 520 (Mon-Fri 9 AM-5 PM CET)

Email: [email protected]

Web: http://www.ibbl.lu

Round: PBMC16R1

Preliminary ; Final ; Amended

Date of Round Closure: 21st October 2016

Report Number: PBMC16R1_Report01

Comments (reason for amendment or re-issue):

Participant’s name: Okayama University Hospital Biobank

Participant Code: L261

mailto:[email protected]

http://www.ibbl.lu/



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SECTION 1 - General Information

Design of the Viable PBMC Isolation Scheme: The Participant proceeded to the collection of one blood tube (any anti-coagulant permitted) from a healthy donor. The Participant isolated the PBMC following his/her routine procedure. The Participant measured the number of PBMC (cells/mL) and prepared up to five PBMC aliquots. Expected viability and cell concentration are the following: Viability: minimum 80%

Cell concentration: between 5*106 and 107 cells/mL of medium.

The Participant proceeded to the freezing of the PBMC aliquots following his/her routine procedure. The isolated PBMC aliquots were shipped back to the PT provider on dry ice in labelled tubes and related data were reported by the Participant through the website http://biospecimenpt.ibbl.lu.

The PBMC viability, early apoptosis/necrosis, number of viable PBMC isolated per mL of blood and PBMC functionality, measured by ELISPOT, for the total PBMCs isolated by each participant, were assessed by IBBL.

Confidentiality: The identities of Participants and their results and performance are kept confidential and are known only to persons involved in the operation of the PT Scheme.

Subcontractors: IBBL is implementing this Program internationally in collaboration with ISBER, the International Society for Biological and Environmental Repositories, based at 375 West 5th Ave, Suite 201, Vancouver, BC, Canada V5Y 1J6. The ISBER PT Advisory Group participates in the development of the annual PT program, in the Value Assignment of the Test Items, and provides advice to PT Participants.

Technical data: For your reference, all information related to the description of the Processing Item is included in the “Processing Item Information Sheet” (PIIS) which was provided to you along with the sample. Extra copies can be downloaded on IBBL’s website (http://www.ibbl.lu/ibbl-services/pt/proficiency-testing-schemes/).

Personalized Advice Service: Advice and educational feedback to Participants is available on demand (please contact IBBL using details on page 1 of this report).

Customer Satisfaction Survey: IBBL seeks the feedback of all Participants, both positive and negative, to be used and analysed to improve its management system, future Schemes, and customer service. Please complete our survey, available online: https://www.surveymonkey.com/r/IBBL2016PT

http://biospecimenpt.ibbl.lu/

http://www.ibbl.lu/ibbl-services/pt/proficiency-testing-schemes/

http://www.ibbl.lu/ibbl-services/pt/proficiency-testing-schemes/

https://www.surveymonkey.com/r/IBBL2016PT



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SECTION 2 – General Statistics

31 Participants registered for the Viable PBMC Isolation Round performed in 2016.

Geographic Origin of Participants

Country Count %

BELGIUM 2 6

ITALY 1 3

JAPAN 3 10

LUXEMBOURG 1 3

SINGAPORE 2 6

SOUTH AFRICA 2 6

SPAIN 1 3

UNITED STATES 19 61

Type of Facilities

Facility Count %

not specified 21 68

Hospital Laboratory 5 16

Private Laboratory 4 13

University/Academic 1 3



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Anticoagulant used for the blood draw

Anticoagulant Count %

Cell Preparation Tube (CPT) - Na2 Citrate 1 3

Cell Preparation Tube (CPT) - Na2 Heparin 1 3

Citrate dextrose additives (ACD) 5 16

K2 EDTA 11 35

Lithium Heparin 1 3

Sodium Heparin 12 39

Extraction equipment

Extraction Count %

Automatic extraction 1 3

Manual extraction 30 97

Extraction tube

Tube Count %

not specified 1 3

Cell Preparation Tube (CPT) 2 6

LeucoSep Tube 2 6

Other - Please Specify 2 6

SepMate Tube (StemCell) 6 19

Standard conical tube 18 58



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Cryopreservation medium

Medium Count %

Commercial 10 32

Home made 21 68

Cryopreservation method

Method Count %

not specified 1 3

Mr Frosty 14 45

Other - Please Specify 13 42

Progressive Rate Freezer 3 10

Storage temperature after aliquoting

Temperature Count %

-80°C 15 48

Liquid phase of LN2 2 6

Vapor phase of LN2 14 45



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SECTION 3 – Data Analysis Results and Evaluation of Performance

Statistical Procedure Used

Basic statistical procedures were used to calculate the PBMC extraction efficiency of the Participants. The Isolated PBMC Viability (%), the Isolated PBMC Apoptosis/Necrosis (%), the Isolated Viable PBMC Yield (cells/mL of blood) and Isolated PBMC functionality (ELISPOT) were calculated. For each parameter, the Participants’ mean was considered as the assigned value. The standard deviation was the standard deviation of the Participants. The outliers were calculated with the Grubb’s test, and were excluded from the statistical summaries.

The scoring system is based on deviation from the assigned value.

1. For the Isolated PBMC Viability and the Isolated Viable PBMC Yield, the scoring system is the following:

Deviation from assigned value (z-score) Consensus Score

< –3 standard deviation 3



> -1 and < +1 standard deviation 0

> +1 standard deviation 0

This scoring system highlights the proficiency of Participants to obtain higher yield and viability of cells than the consensus mean.

2. For the Isolated PBMC Apoptosis/Necrosis, the scoring system is the following:

Deviation from assigned value (z-score) Consensus Score


> -1 and < +1 standard deviation 0




This scoring system highlights the proficiency of Participants to obtain lower quantity of apoptotic/necrotic cells than the consensus mean.



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Isolated Viable PBMC Yield per mL of Blood

YOU SUBMITTED RESULTS WITH THIS METHOD

Expected Results

Panel Composition and expected result (mean values and participant data standard deviations)

Sample Sample content

Mean (10^6 ç/ml blood)

Proficiency testing standard deviation (10^6 ç/ml blood)

All Results

PBMC 1.01 0.78

Your Results

Sample Sample content Your result (10^6 ç/ml blood) Your consensus score

Your Aliquot(s) PBMC 0.37 0

Sum quantitative panel score 0

Your z-score for Isolated Viable PBMC Yield per mL of Blood is -0.821 (Your Aliquot(s)).

Your z-score for Isolated Viable PBMC Yield per mL of Blood has been designated as "accurate" or "very satisfactory".

If your z-score has been designated as "questionable" or "requiring action", some additional feedback may be found at the end of this report.

Your results are shown in green.

Your Aliquot(s) Results

PROLab Plus

Laboratory

L100

L226

L205

L261

L238

L216

L018

L046

L108

L150

L234

L056

L137

L065

L255

L267

L223

L217

L242

L124

L186

L203

L037

L066

L101

L028

L253

L219

L228

L230

L2141

0^6

ç/m

l blo

od

5

0

No. of laboratories: 30No. of outlier values: 1Mean: 1.01 10^6 ç/ml bloodProficiency Testing Standard Deviation: 0.78 10^6 ç/ml blood (Empirical value)

Median: 0.78Minimal value: 0.16Maximal value: 3.98Range: 3.82

B



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Your History of Z-Scores

This is your first participation with this method in the scheme; therefore, no history of your z-scores is available yet.

A history of your z-scores will be available in your future participations.



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Isolated PBMC Viability


Expected Results


Sample Sample content Mean (%) Proficiency testing standard deviation (%)

All Results PBMC 85.2 4.9

Your Results

Sample Sample content Your result (%) Your consensus score



Your z-score for Isolated PBMC Viability is 0.157 (Your Aliquot(s)).

Your z-score for Isolated PBMC Viability has been designated as "accurate" or "very satisfactory".




PROLab Plus

Laboratory

L056

L066

L186

L234

L124

L214

L137

L108

L226

L100

L238

L267

L255

L223

L253

L261

L228

L046

L217

L230

L065

L028

L101

L203

L205

L150

L216

L037

L018

L219

L242

%

100

80

60

No. of laboratories: 30No. of outlier values: 1Mean: 85.2 %Proficiency Testing Standard Deviation: 4.9 % (Empirical value)


B



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Isolated PBMC Apoptosis/Necrosis


Expected Results


Sample Sample content Mean (%) Proficiency testing standard deviation (%)

All Results PBMC 14.95 4.83

Your Results

Sample Sample content Your result (%) Your consensus score



Your z-score for Isolated PBMC Apoptosis/Necrosis is -0.154 (Your Aliquot(s)).

Your z-score for Isolated PBMC Apoptosis/Necrosis has been designated as "accurate" or "very satisfactory".




PROLab Plus

Laboratory

L242

L219

L018

L037

L216

L150

L203

L205

L101

L065

L028

L230

L217

L046

L228

L261

L253

L223

L255

L267

L238

L100

L226

L108

L137

L214

L124

L234

L186

L066

L056

%

50

0

No. of laboratories: 30No. of outlier values: 1Mean: 14.95 %Proficiency Testing Standard Deviation: 4.83 % (Empirical value)


B



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IFN-gamma ELISPOT assay

High spot count (more than 20 spots/200000 cells for CPI)

Low spot count (less than 20 spots/200000 cells for CPI)

CPI Antigen - a pool of protein antigens derived from Cytomegalovirus, Influenza, and Parainfluenza viruses, used as a positive control for the functionality of CD4+ T cells and antigen presenting cells

IFN-gamma ELISPOT assay Number of Participants Your result

High spot count 30 X

Low spot count 1

IFN-gamma ELISPOT assay

High spot count Low spot count

Num

ber

of part

icip

ants

30

28

26

24

22

20

18

16

14

12

10

8

6

4

2

0



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Global Viable PBMC Isolation 2016 Z-Scores For Your Aliquot(s)

PROLab Plus

Z-Score

-3 0 3 -3 0 3 -3 0 3 -3 0 3

Method

IFN

-gam

ma E

LIS

PO

T a

ssay

Isola

ted P

BM

C V

iability

Isola

ted P

BM

C A

popto

sis

/Necro

sis

Isola

ted V

iable

PB

MC

Yie

ld p

er

mL o

f B

lood

Labora

tory

7.120-7.049

3.817

3.033

3.110



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Overview Of Your Z-Scores For All Testing Schemes

PROLab Plus

Z-Score

-3 0 3 -3 0 3 -3 0 3

Round/Sample

DN

AQ

16R

1/T

ube A

DN

AQ

16R

1/T

ube B

DN

AQ

16R

1/T

ube C

Meth

od

DNA Concentration - Spectrofluorometry



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Overview Of Your Z-Scores For All Processing Schemes

PROLab Plus

Z-Score

-3 0 3 -3 0 3 -3 0 3 -3 0 3 -3 0 3 -3 0 3

Round/SampleD

NA

BLD

16/A

ll re

sults

DN

AB

LD

16/S

ilica m

em

bra

ne-b

ased

CF

DN

A16/A

ll re

sults

CF

DN

A16/S

ilica-m

em

bra

ne b

ased

PB

MC

16R

1/A

ll R

esults

PB

MC

16R

1/Y

our

Aliq

uot(

s)

Meth

od

PROLab Plus


proficiency testing for the quality control

of biospecimens

This report has been validated by the PT Coordinator.

Fay BETSOU, PhD HDR, Coordinator

Documents

PROFICIENCY TESTING REPORT - 岡大バイオバン …biobank.ccsv.okayama-u.ac.jp/.../01/PBMC16R1-L261-Report.pdfservices/pt/proficiency-testing-schemes/). Personalized Advice Service: