Scheme 1B - 2017-03-04

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UK NEQAS FOR H&I SCHEME 1B - HLA-B27 TESTING HLA-B27 TYPING RESULTS OF SAMPLES 1B03&04/2017 DESPATCHED ON 25/04/2017 Summary of Results Total tested (excl. Equivocal) Positive Negative NT/Equivocal % Positive % Negative

122

118

Donor HLA type

1B03/2017

HLA-A1, A24; B7, B44; Cw4, Cw7; DR13, DR15, DQ6, DQ-

1 121 4 0.8% 99.2%

117 1 8 99.2% 0.8%

Donor HLA type

1B04/2017

HLA-A2, A-; B27, B62; Cw2, Cw10; DR1, DR12, DQ5, DQ7

Negative

Positive

Result 1B03/2017

Result 1B04/2017

Date received

Date tested

Method

1 3 4 7 8 9 10 11 12 14 15 17 19 20 21 22 23 24 25 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 72 76 77 78 83 84 85 86 90 96 101 103 104 106 107 108 109 110 111 112 113 115 126 127 128 129 130 133 137

Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Positive 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 Equivocal

Positive Positive Positive Positive Equivocal Positive Positive Positive Positive Positive Positive Positive Positive Positive Negative Positive Positive Positive Positive Positive Equivocal Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive NT Equivocal Positive Positive Positive Positive Positive Positive Equivocal Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive

27-Apr 26-Apr 26-Apr 27-Apr 26-Apr 26-Apr 26-Apr 26-Apr 26-Apr 11-May 26-Apr 02-May 27-Apr 28-Apr 26-Apr 26-Apr 26-Apr 28-Apr 27-Apr 26-Apr 26-Apr 26-Apr 26-Apr 27-Apr 26-Apr 26-Apr 26-Apr 28-Apr 26-Apr 26-Apr 26-Apr 26-Apr 27-Apr 26-Apr 27-Apr 27-Apr 26-Apr 23-Apr 26-Apr 26-Apr 27-Apr 26-Apr 26-Apr 26-Apr 28-Apr 26-Apr 27-Apr 26-Apr 26-Apr 26-Apr 23-May 27-Apr 26-Apr 27-Apr 26-Apr 26-Apr 27-Apr 27-Apr 05-May 27-Apr 26-Apr 26-Apr 27-Apr 27-Apr 26-Apr 26-Apr 02-May 26-Apr 26-Apr 03-May 26-Apr 27-Apr 26-Apr

08-May 26-Apr 08-May 27-Apr 28-Apr 28-Apr 26-Apr 03-May 03-May 15-May 02-May 03-May 28-Apr 02-May 27-Apr 28-Apr 27-Apr 03-May 03-May 27-Apr 26-Apr 02-May 26-Apr 02-May 26-Apr

Molecular Serological Molecular Serological Serological Molecular Serological Molecular Molecular Molecular Molecular Molecular, Serological Molecular Serological Serological Molecular Molecular Molecular Molecular Molecular Serological Molecular Molecular Serological Serological Molecular

142 153 154 156 174 176 185 198 209 219 223 224 225 232 234 236 245 252 255 256 257 269 273 276 278 279 288 292 295 300 305 306 307 310 317 319 324 325 326 327 329 334 339 340 349 353 357 359 362 365 368 370 372

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 NT Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative

Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive NT NT Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Equivocal Positive Positive Positive

04-Apr 26-Apr 26-Apr 26-Apr 01-May 02-May 26-Apr 27-Apr 26-Apr 22-May 26-Apr 26-Apr 02-May 04-May 02-May 03-May 26-Apr 29-Apr 26-Apr 28-Apr 26-Apr

Consensus Lab No

26-Apr 04-May 27-Apr 27-Apr 05-May 02-May 02-May 28-Apr 27-Apr 02-May 04-May 08-May 27-Apr 28-Apr 27-Apr 03-May 26-Apr 26-Apr NT 27-Apr 27-Apr 27-Apr 27-Apr 26-Apr 25-May 02-May 26-Apr 05-May 28-Apr 28-Apr 27-Apr 28-Apr 05-May 02-May 05-May 26-Apr 27-Apr 03-May 03-May 26-Apr 08-May 28-Apr 02-May 04-May 27-Apr 28-Apr 26-Apr

High Resolution Typing HLA-B*27 alleles 1B03/2017 1B04/2017

1B 04/2017 is equivocal as the result was 22.6. Indeterminant zone is 15 to 23

Report to replace previously submitted results

B*27:03/27:05

Molecular Molecular Serological Molecular Molecular Molecular Serological Serological Molecular Molecular Molecular Serological Molecular Molecular Molecular Serological Serological NT Serological Serological Molecular Serological Molecular, Serological

B*27:03+ 1B04/2017 falls close to decision line and would be sent for confirmation

This test is not up and running in yet as we are awaiting staffing. The results from both analysers fall within the Equivocal range for the assay.

Molecular Serological Molecular Molecular Serological Serological Molecular Molecular Molecular Molecular, Serological Serological Molecular Molecular Molecular Molecular Molecular Molecular, Serological Molecular Molecular, Serological Serological Serological

Molecular Serological Molecular Molecular Molecular Serological Molecular Serological Molecular

26-Apr 26-Apr 02-May 26-Apr 26-Apr 27-Apr 26-Apr NT

03-May 28-Apr 05-May 03-May 08-May 02-May 27-Apr 27-Apr 03-May 08-May 02-May 04-May 05-May 05-May 05-May 04-May 02-May 03-May 05-May 28-Apr 08-May 03-May 28-Apr 03-May 08-May 29-Apr 27-Apr 27-Apr 28-Apr NT

26-Apr 26-Apr 26-Apr 02-May 26-Apr

27-Apr 02-May 26-Apr 03-May 27-Apr

26-Apr 27-Apr 02-May 26-Apr 27-Apr 26-Apr 26-Apr 27-Apr 26-Apr 27-Apr 18-Apr 26-Apr 27-Apr 26-Apr 28-Apr 24-Apr

08-May 27-Apr 03-May 27-Apr 27-Apr 26-Apr 28-Apr 28-Apr 02-May 27-Apr 02-May 03-May 03-May 28-Apr 28-Apr 03-May

Molecular Molecular Serological Molecular Molecular Serological Molecular Serological Molecular Serological Serological Molecular Molecular Serological Molecular Serological Molecular Molecular Serological Molecular Serological Serological

Molecular Molecular Molecular Molecular Molecular Serological Molecular Molecular Molecular Serological Molecular Molecular Molecular Molecular Molecular Serological Serological Serological Serological NT

Comments

Result falls within indeterminate range and would be referred for confirmatory testing.

Negative in exon 2 and 3

Positive in exon 2 and 3

Sample 1B03 was HLA-B27 positive and HLA-B7 positive. Hence, it is possible that HLA-B27 positivity of sample 1B03 is a result of cross reaction (ABCm3 clone). Sample 1B03 was B27 positive when it was tested with an antibody against an alternative epitope (MPC-11 clone).

Sample 1B03 showed B7 cross reaction

Exon 2 & Exon 3

Samples not received as of 19/05/2017 No results returned

Positive diagnosis with our technique, must be confirmed by reference technique

UK NEQAS for H&I is operated by Velindre NHS Trust, a UKAS proficiency testing provider No 8351. © Confidential report; no data may be published without permission from UK NEQAS for H&I

NEQ-115 Issue 2 P. 1 of 2 Effective Date 10/10/16

UK NEQAS FOR H&I SCHEME 1B - HLA-B27 TESTING HLA-B27 TYPING RESULTS OF SAMPLES 1B03&04/2017 DESPATCHED ON 25/04/2017 Lab No

Source of pos/neg control cells used (if applicable) 17 20 Control Cells: B7 Homozygous Cell, B27/B7 Positive cell, B27/B7 Negative cell 21 27

Positive control: Crisp HLA-B27 positive control cells from Phoenix Flow Systems Negative control: ImmunoTrol cells from Beckman Coulter 31 CRISP Cells

68 110 111 128

Positive Controls -Crisp HLA-B27 Supplier- Phoenix Flow Systems N/A

Monoclonal 1 Designation Specificity B27 FITC B27 B27-FITC, B27 and B7, Becton Dickinson

Manufacturer One Lambda

HLA-B27-FITC One Lambda

Monoclonal 2 Designation Specificity CD3 PE CD3 B27-FITC, B27, One Lambda

Manufacturer BD

Designation

Monoclonal 3 Specificity

Manufacturer

N/A

HLA-B27-FITC / HLA-B7-PE Beckman Coulter

Beckman Coulter Duraclone

IgG2a-FITC, IgG1-PE, Isotype Control, Beckman Coulter HLA B27-FITC/HLA B7-PE, Beckman Coulter Anti HLA B27 FITC Anti HLA B27 FITC IOTEST HLA-B27-FITC, HLA-B27, Beckman coulter

133 137 324

In house

Anti CD27 HLA-ABCm3/BB71 Anti HLA B27 FITC

365

Internal control in each Duraclone B27 tube (Bc) (beads)

HLA-B27 (clone FD 705)

DuraClone B27, HLA-B27, Beckman Coulter

IOTest HLA-B27-FITC HLA-B7PE, HLA-B27 and HLA-B7, Beckman Coulter HLA B27-FITC, One Lambda FD705-9E1E10 Clone GS145.2

One Lambda BD Biosciences

CD27 BC HLA-B27/B7 Immunotech Anti HLA B27 FITC Beckman Coulter PE

Beckman Coulter

UK NEQAS for H&I is operated by Velindre NHS Trust, a UKAS proficiency testing provider No 8351. © Confidential report; no data may be published without permission from UK NEQAS for H&I

CD3 PE IOTEST HLA-B7-PE, HLA-B7, Beckman coulter Anti CD7 MPC-11 HLA-B7-PE HLA-B27 (clone HLA-ABC-m3)

Clone SK7

BD Biosciences CD45 PerCP-Cy5.5, Lymphocytes Gating, Becton Dickinson

CD7 BC HLA-B27 One Lambda HLA-B7-PE Beckman Coulter PE

Beckman Coulter

Anti CD3

CD3 (clone UCHT1)

T cells

BC

PE-Dyomics 649 Beckman Coulter

NEQ-115 Issue 2 P. 2 of 2 Effective Date 10/10/16