Scheme 1B-2018-03-04

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UK NEQAS FOR H&I SCHEME 1B - HLA-B27 TESTING

HLA-B27 TYPING RESULTS OF SAMPLES 1B03&04/2018 DESPATCHED ON 15 MAY 2018 Summary of Results Total tested (excl. Equivocal) Positive Negative NT/Equivocal % Positive % Negative

124

123

Donor HLA type

1B03/2018

HLA-A3, A30; B7, B13; Cw6, Cw7; DR7, DR9, DQ2, DQ9

0 124 2 0.0% 100.0%

123 0 3 100.0% 0.0%

Donor HLA type

1B04/2018

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

Negative

Positive

1 3 4 7 8 9 10 11 12 14 15 17 19 20 21 22 23 24 25 26 28 29 31 32 34 35 37 39 40 41 42

Result 1B03/2018 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

Result 1B04/2018 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 Positive Positive

Date received 16-May 16-May 16-May 16-May 17-May 16-May 18-May 15-May 22-May 16-May 17-May 17-May 17-May 16-May 21-May 16-May 29-May 17-May 16-May 16-May 16-May 16-May 16-May 21-May 16-May 16-May 17-May 17-May 17-May 16-May 17-May

Date tested 18-May 16-May 23-May 17-May 17-May 18-May 18-May 23-May 30-May 22-May 21-May 24-May 18-May 17-May 21-May 17-May 30-May 21-May 21-May 16-May 22-May 16-May 16-May 24-May 21-May 18-May 23-May 17-May 17-May 23-May 22-May

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

45 46 47 48 54 55 57 58 62 65

Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative

Positive Positive Positive Positive Positive Positive Positive Positive Positive NT

18-May 16-May 18-May 17-May 17-May 16-May 17-May 17-May 16-May 24-May

23-May 18-May 18-May 21-May 18-May 22-May 18-May 18-May 16-May 28-May

Molecular Serological Serological Molecular Molecular Molecular Serological Molecular Molecular Serological

66 67 72 76 77 78 83 84 85 86 90 96 101 103 104 106 107

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

17-May 16-May 18-May 16-May 16-May 17-May 16-May 16-May 17-May 16-May 21-May 16-May 16-May 16-May 18-May 17-May 22-May

25-May 16-May 24-May 16-May 17-May 18-May 18-May 18-May 20-May 17-May 22-May 22-May 17-May 17-May 18-May 18-May 25-May

Molecular Serological Molecular Serological Serological Molecular Serological Serological and Molecular Molecular Molecular Serological Molecular Molecular Serological Serological Molecular

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 Equivocal

Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive

19-May 16-May 18-May 16-May 18-May 16-May 16-May 24-May 16-May 17-May 17-May 16-May 18-May 18-May

24-May 17-May 18-May 18-May 24-May 22-May 18-May 28-May 22-May 05-Jun 18-May 17-May 22-May 22-May

Molecular Molecular Serological and Molecular Serological Molecular Molecular Molecular Molecular Molecular Serological and Molecular Molecular Serological and Molecular Serological and Molecular Serological

142 150 153 154 156 159 174 176 185

Negative Negative Negative Negative Negative Negative Negative Negative Negative

Positive Positive Positive Positive Positive Positive Positive Positive Positive

16-May 16-May 16-May 16-May 16-May 16-May 28-May 22-May 16-May

18-May 16-May 18-May 22-May 23-May 18-May 29-May 22-May 16-May

Molecular Molecular Serological Molecular Molecular Molecular Molecular Serological Molecular

198 209 219 223 224 225 232 234 236 245 252 255 256 257 269 273 276 278

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

17-May 16-May 24-May 16-May 16-May 24-May 19-May 20-May 12-May 16-May 17-May 16-May 16-May 16-May 16-May 16-May 24-May

18-May 18-May 25-May 23-May 23-May 29-May 23-May 29-May 18-May 23-May 17-May 24-May 16-May 24-May 24-May 25-May 23-May 28-May

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

279

Negative

Positive

16-May

21-May

Serological and Molecular

288 292 295 300 305 306 307 315 317 319 324 327 334 339 340 348 349 353 357 359 362 365 368 372 373 377 378 401

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

Positive Positive Positive NT Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive — Equivocal Positive Positive Positive Positive Positive Positive Positive — Positive Positive Positive

16-May 17-May 16-May NT 10-May 16-May 17-May 16-May 23-May 16-May 17-May 16-May 16-May 16-May 17/05/2018 — 17/05/2018 17/05/2018 17/05/2018 16/05/2018 16/05/2018 16/05/2018 16/05/2018 17/05/2018 — 21/05/2018 21/05/2018 30/05/2018

16-May 17-May 18-May NT 25-May 17-May 21-May 18-May 28-May 16-May 17-May 17-May 18-May 17-May 22-May — 17-May 17-May 17-May 24-May 16-May 16-May 28-May 22-May — 21-May 21-May 01-Jun

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

Consensus

Lab No

Method

PCR-SSP Detection system

Primers and Probes

Own design Own design Own design

Gel Gel Gel

Commercial kits

Fluorescence

Own design Own design

Gel Gel

Commercial kits

Gel, BAG reverse dot blot

1B03/2018

High Resolution Typing HLA-B*27 alleles 1B04/2018

Comments

B*27- only defined to first field B*27:03/05 allele group

B*27:03/05/05Q/13

Commercial kits

SSOP

Own design

Gel

Own design

Gel

Own design

Gel

Commercial kits Own design

Gel Gel

Commercial kits

Gel

27:03/05/05Q/13/17/19/55/56/58/60/61/64N/67/70 /72/74/80/82/84/87/88/90/93/94N/9699/104/110/111/117/118/121/123/124/128/131/13 5/140/141/143-146/150/152/155/159/162 B*27:05/47/90/116-162

Sample 1B 04/2018 result by flow cytometry fell within the manufacturer's recommended equivocal range. Our laboratory procedure is to send samples to an external laboratory for testing by PCR in these cases. Instructed by NEQAS to return these results as not tested rather than equivocal.

18B061392 18B061393 Commercial kits

Fluorescence

Commercial kits

Gel

Commercial kits Commercial kits

Gel Gel

Commercial kits Commercial kits

Fluorescence Luminex

Commercial kits

Gel

Own design

Fluorescence

Commercial kits Own design

Gel Fluorescence

Commercial kits

Gel

Commercial Commercial Commercial

Coloured dots by substrate conversion on microchip Gel

Commercial kits

Fluorescence

Commercial

Melting point

1B032018 was Negative in Exon 2 and Exon 3. 1B042018 was positive in Exon 2 and exon 3

Sample 1B03 was positive with HLA-B27 (clone: HLA-ABCm3) and HLA-B7 (clone:BB7.1) antibodies, however, it was negative with the second HLA-B27 (clone:FD705) antibody.

*27:01-27:05:08, 27:05:10-27:05:15, 27:05:1727:05:22, 27:05:24-27:05:32, 27:08, 27:10, 27:1227:13, 27:15-27:17, 27:25, 27:28, 27:36-27:40, 27:42, 27:44-27:45, 27:47-27:69, 27:71-27:74, 27:79-27:80, 27:82-27:84, 27:86-27:90:03, 27:9227:100, 27:103-27:105, 27:108-27:124, 27:126, 27:128, 27:131-27:135, 27:137, 27:137, 27:139, 27:141-27:149, 27:151-27:152, 27:155-27:156

Sample 1B 03/2018 negative for exon 2 and exon 3 Sample 1B 04/2018 positive for exon 2 and exon 3 The serological method used for each sample is: CDC-N,IH The molecolar typing method used for each sample is: SSP-L.R.

Samples not received.

Commercial Commercial

Fluorescence Tapestation (Agilent)/Gel

Commercial

Gel

Not participating in 1B03&04/18 Serological Molecular Serological Molecular Molecular Serological Molecular Serological

in house HLA-B27

NEQ-115

Not participating in 1B03&04/18 Serological Serological Molecular

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

Issue 2 P. 1 of 1 Effective Date 10/10/16