Scheme 1B - 2016-01-02

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UK NEQAS FOR H&I SCHEME 1B - HLA-B27 TESTING HLA-B27 TYPING RESULTS OF SAMPLES 1B01&02/2016 DESPATCHED ON 5TH JANUARY 2016

Summary of Results Total tested (excl. Positive

115 0

114 0

Donor HLA type

1B01/2016

HLA-A2, A24; B57, B60; Cw6, Cw10; DR7, DR8; DQ4, DQ9

Negative

115

114

Donor HLA type

1B02/2016

HLA-A1, A31; B8, B60; Cw7, Cw10; DR4, -; DQ7, DQ8

Method

Source of pos/neg control cells used (if applicable)

NT/Equivocal % Positive % Negative Consensus Lab No 1 3 4 7 8 9 10 11 12 14 15 16 17 19 20 21 22 23 24 26 27 28 29 31 32 34 35 37 39 40 41 42 45 46 47 48 54 55 57 58 62 66 67 68 71 72 78 83 84 85 86 90 93 96 101 103 104 106 107 108 109 110 111 112 113 115 126 127 128 129 130 131 137 142 153 154 156 158 174 176 185 198 199 209 219 223 225 232 234 245 253 255 256 257 269 273 276 278 279 285 288 292 295 300 305 306 307 310 317 319 323 324 325 326 327 329 334 339 340

4

5

0.0%

0.0%

100.0%

100.0%

Negative

Negative

Result 1B01/2016 Negative Negative Negative Negative NT Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative NT Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative NT Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative NT

Result 1B02/2016 Negative Negative Negative Negative NT Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Equivocal Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative NT Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative NT Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative NT

Date received 07-Jan 07-Jan 06-Jan 05-Jan NT 06-Jan 06-Jan 04-Jan 07-Jan 06-Jan 07-Jan 07-Jan 13-Jan 06-Jan 06-Jan 06-Jan 07-Jan 07-Jan 07-Jan 07-Jan 06-Jan 06-Jan 06-Jan 06-Jan 06-Jan 06-Jan 06-Jan 06-Jan 11-Jan 07-Jan 07-Jan 07-Jan 08-Jan 11-Jan 07-Jan 07-Jan 07-Jan 16-Jan 06-Jan 06-Jan 06-Jan 06-Jan 06-Jan 06-Jan 06-Jan 11-Jan NT 08-Jan 07-Jan

Date tested 11-Jan 08-Jan 08-Jan 06-Jan NT 07-Jan 06-Jan 12-Jan 12-Jan 07-Jan 08-Jan 12-Jan 21-Jan 08-Jan 06-Jan 07-Jan 07-Jan 13-Jan 14-Jan 07-Jan 07-Jan 12-Jan 06-Jan 07-Jan 06-Jan 07-Jan 08-Jan 14-Jan 13-Jan 08-Jan 14-Jan 12-Jan 08-Jan 13-Jan 07-Jan 12-Jan 08-Jan 25-Jan 06-Jan 11-Jan 06-Jan 12-Jan 06-Jan 07-Jan 22-Jan 20-Jan NT

07-Jan 08-Jan 06-Jan 06-Jan 06-Jan 06-Jan 08-Jan 08-Jan 12-Jan 08-Jan 06-Jan 07-Jan 06-Jan 08-Jan 06-Jan 08-Jan 11-Jan 07-Jan 06-Jan 14-Jan 07-Jan 06-Jan 06-Jan 05-Jan 14-Jan 06-Jan 15-Jan 08-Jan 09-Jan 04-Jan 06-Jan 07-Jan 11-Jan 06-Jan 14-Jan 07-Jan 12-Jan NT 11-Jan 06-Jan 11-Jan 07-Jan 06-Jan 11-Jan

08-Jan 08-Jan 07-Jan 18-Jan 12-Jan 08-Jan 08-Jan 12-Jan 15-Jan 13-Jan 11-Jan 08-Jan 07-Jan 14-Jan 07-Jan 13-Jan 18-Jan 08-Jan 06-Jan 18-Jan 08-Jan 08-Jan 06-Jan 14-Jan 14-Jan 14-Jan 19-Jan 11-Jan 12-Jan 11-Jan 07-Jan 08-Jan 14-Jan 08-Jan 22-Jan 12-Jan 20-Jan NT 12-Jan 08-Jan 25-Jan 12-Jan 06-Jan 11-Jan

06-Jan 06-Jan 25-Jan 07-Jan 07-Jan 07-Jan 07-Jan 07-Jan 12-Jan 17-Jan 06-Jan 11-Jan 05-Jan 11-Jan 06-Jan 07-Jan 07-Jan 07-Jan 08-Jan 15-Jan 18-Jan 28-Jan 06-Jan NT

13-Jan 08-Jan 25-Jan 09-Jan 07-Jan 08-Jan 07-Jan 08-Jan 13-Jan 22-Jan 07-Jan 25-Jan 07-Jan 12-Jan 13-Jan 07-Jan 07-Jan 18-Jan 08-Jan 25-Jan 22-Jan 28-Jan 12-Jan NT

07-Jan

PCR-SSP Flow Cytometry PCR-SSP Flow Cytometry

Enzo Life Sciences

PCR-SSP

In-house

Primers and Probes Primers - Alta Bioscience Gel

PCR-SSP Detection system

Own design

Fluorescence, SYBR GREEN and Melting Curve

Own design

Gel

Own design Own design

Gel Gel

Olerup Invitrogen

Gel Gel

Own design Olerup

Gel Gel

Biofortuna

Gel

Olerup Own design Own design Own design

Gel Gel Gel, Fluorescence Gel

High Resolution Typing HLA-B*27 alleles 1B01/2016 1B02/2016

Comments

No results returned

Luminex PCR-SSP PCR-SSP Luminex In-house, Commercial PCR-SSP Flow Cytometry, PCR-SSP In-house Flow Cytometry Flow Cytometry PCR-SSP PCR-SSP Flow Cytometry PCR-SSO PCR-SSP/SSO PCR-SSP/SSO Flow Cytometry PCR-SSP PCR-SSP PCR-SSP PCR-SSP Flow Cytometry Luminex PCR-SSP Flow Cytometry Flow Cytometry PCR-SSP PCR-SSP PCR-SSP Flow Cytometry PCR-SSP PCR-SSP, SBT RT-PCR - TAQMAN Flow Cytometry Flow Cytometry Flow Cytometry PCR-SSP PCR-SSP Flow Cytometry Flow Cytometry, PCR-SSP PCR-SSP Flow Cytometry Flow Cytometry PCR-SSP PCR-SSP Flow Cytometry Flow Cytometry

PCR-SSO, BAG Healthcare, reverse dot-blot assay

Commercial

In-house Axxora In-house

Own design

Gel

Invitrogen Own design Hain Lifesciences

Gel Gel Fluorescence

Innotrain Olerup Own design

Gel Fluorescence

Fluorescence Commercial

Gel

Olerup

Gel

All except B*27:05:23 and B*27:75

All except B*27:05:23 and B*27:75 Unknown whether B*27:91 and B*27:93 will be detected

In-house Secore

Crisp HLA-B27 TIB MOLBIOL Enzo Life Sciences Enzo Life Sciences In-house Beckman+One Lambda Olerup

Invitrogen

Gel

Olerup Olerup

Gel Gel

EuroImmun

Fluorescence

Commercial

Gel

Own design Olerup Olerup

Gel Gel Gel

Olerup

Gel

Olerup

Gel

Olerup Gel

Gel

Olerup

Gel

Linkafe Biosciences Commercial

Fluorescence Gel

Invitrogen Olerup

Gel Gel

Samples not received

B*27:01 to B*27:135

B*27:01 to B*27:135

Exon 2 and Exon 3

Exon 2 and Exon 3

Phoenix

PCR-SSO Flow Cytometry, PCR-SSP Flow Cytometry In-house PCR-SSP PCR-SSP PCR-SSP

RT-PCR PCR-SSP

B*27:01 to B*27:138

Enzo Life Sciences In-house

PCR-SSP

EuroArray Flow Cytometry, Luminex PCR-SSO Cytotoxicity, PCR-SSP Flow Cytometry Flow Cytometry RT-PCR Flow Cytometry PCR-SSP RT-PCR PCR-SSP PCR-SSP/SSO, Microarray Cytotoxicity PCR-SSP

B*27:01 to B*27:138

In-house

Phoenix 1B02 was slightly pos with monoclonal 1, but neg with monoclonal 2 In-house, Commercial

Invitrogen In-house

UK NEQAS 1B

PCR-SSP PCR-SSP NT

Samples not released from customs

PCR-SSO RT-PCR RT-PCR Flow Cytometry PCR-SSP

Commercial

PCR-SSP PCR-SSP

Cytotoxicity Flow Cytometry Cytotoxicity Flow Cytometry PCR-SSP PCR-SSP PCR-SSP Flow Cytometry PCR-SSP RT-PCR Cytotoxicity Flow Cytometry PCR-SSP

PCR-SSO Flow Cytometry PCR-SSP NT

Olerup EuroImmun Olerup Olerup

Gel Gel

Gel

Innotrain

Gel

Sciences Biomark inc Olerup

Fluorescence Gel

Olerup

Gel

BAG Healthcare

Gel

In-house

In-house In-house

AnDiaTec

Fluorescence

Own design

Fluorescence Analysis not performed - Method still under validation

Page 1 of 2 UK NEQAS for H&I is provided from the Welsh Blood Service, a division of Velindre NHS Trust © Copyright Notice: Reports are confidential, no data may be published without permission from UK NEQAS for H&I

1B v1 12/08/15

UK NEQAS FOR H&I SCHEME 1B - HLA-B27 TESTING HLA-B27 TYPING RESULTS OF SAMPLES 1B01&02/2016 DESPATCHED ON 5TH JANUARY 2016 Lab No 3 7 17 20 21 27 31 32 40 46 47 57 67 68 71 83 84 90 93 103 104 110 111 128 131 137 153 256 288 295 310 324 334

Monoclonal 1 Designation HLA-B27 FITC FITC B27 FITC Anti B27-FITC/CD3-PE HLA-B27 Duraclone B27 MSIgG1 RDI/MsIgG2a FITC PF/FITC HLA-B27 PE Anti HLA-B27 ABC m3 IgG2a IgG2a ABC m3 HLA-B27/B7 Duraclone B27 HLA-B27 B7/B27 HLA-B27 B27 HLA-B27-FITC/HLA-B7-PE Anti-HLA-B27 Anti-HLA-B27 FITC Anti-HLA-B27 FITC Iotest HLA-B27 FITC HLA-B27-FITC/HLA-B7-PE HLA-ABC m3/BB7.1 HLA-B27 FITC Anti-HLA-B27 Anti-HLA-B27 Anti-HLA-B27 FITC Iotest HLA-B27 FITC/B7 PE HLA-B27 FITC HLA-B27 FITC

Monoclonal 1 Specificity Clone GS145.2 Anti-HLA-B27 B27 B27+B7 HLA-B27 FITC CD3, HLA-B27, HLA-B7 Isotype control B7/B27 FITC HLA-B27 HLA-B27 HLA-B27 HLA-B27-FITC/B7-PE FITC B7/B27 R-PE B27 HLA-B27 FITC/CD3 PE FD705-9E1E10 Clone GS145.2 HLA-B27 HLA-B27 HLA-B27/B7 IgG2b FD705 HLA-B27 FITC Clone GS145.2 HLA-B27 HLA-B27

Monoclonal 1 Manufacturer BD BD One Lambda BD One Lambda Beckman Coulter Beckman Coulter Beckman Coulter Beckman Coulter BD Beckman Coulter Beckman Coulter Beckman Coulter Beckman Coulter Beckman Coulter Beckman Coulter Beckman Coulter Beckman Coulter One Lambda Beckman Coulter BD One Lambda BD Beckman Coulter Beckman Coulter Immunotech One Lambda BD BD BD Beckman Coulter Beckman Coulter Beckman Coulter

Monoclonal 2 Designation CD3 PE PE/FITC CD3 PE Anti B27-FITC HLA-B27/HLA-B7

Monoclonal 2 Specificity Clone SK7 Anti CD3 CD3 B27 HLA-B27-FITC/HLA-B7-PE

Monoclonal 2 Manufacturer BD BD BD One Lambda Beckman Coulter

HLA-B27 FITC/B7 PE PE/FITC CD3-PE-DY649 Anti CD3 FD705 IgG2b IgG2b FD705 HLA-B27

B7/B27 IgG/IgG2 PE HLA-B27 HLA-B27 HLA-B27 HLA-B27-FITC

Beckman Coulter Beckman Coulter Beckman Coulter BD Beckman Coulter Beckman Coulter Beckman Coulter One Lambda

Monoclonal 3 Designation

Monoclonal 3 Specificity

Monoclonal 3 Manufacturer

Details of Pos/Neg conrol cells used Crisp B7 homozygous control Phoenix flow crisp

BB7.1 IgG1 IgG1K BB7.1

HLA-B7 CD3 HLA-B27

Beckman Coulter Beckman Coulter Beckman Coulter

HLA-B7

PE

Beckman Coulter

CD3

PC5

Beckman Coulter

CD3 B27

PE-Dyomics649 B27

Beckman Coulter Beckman Coulter

B7

Clone BB7.1

Beckman Coulter

CD3 PE Iotest HLA-B7 PE

Clone SK7 HLA-B7

BD Beckman Coulter

CD45 PerCp-Cy5.5

Lymphocytes gating

BD

MPC-11 HLA-B27-FITC/B7-PE Anti-CD3 Anti-CD3 Anti-CD3 PE Iotest HLA-B27 FITC/B7 PE HLA-B7 PE HLA-B7 PE

HLA-B27 IgG2a-IgG1 CD3 PE Clone SK7 HLA-B7 HLA-B7

One Lambda Immunotech BD BD BD Beckman Coulter Beckman Coulter Beckman Coulter

Page 2 of 2 UK NEQAS for H&I is provided from the Welsh Blood Service, a division of Velindre NHS Trust © Copyright Notice: Reports are confidential, no data may be published without permission from UK NEQAS for H&I

Crisp Known positive samples

Crisp

1 Pos, 1 Neg

1B v1 12/08/15