Olerup SSP
IMPROVING RESULTS IN SSP LOW RESOLUTION TYPING
Olerup SSP® Low resolution Kits – Flexibility and Convenience
• A comparative study of different kit formats for: – HLA-A – HLA-B – HLA–DRB
• Objective: – To investigate the frequencies of typing ambiguities for low resolution SSP typings in two sets of samples – To compare the performance of the ”classic” formats to the new formats.
• Material and Methods – 102 randomly selected Swedish blood donors – 102 consecutive UCLA Exchange DNAs – The most current lots of the Olerup SSP® kits for HLA-A, -B and -DRB low resolution typing • ”classic” format vs new, extended formats
– Analyzed using Olerup SSP ® version of Helmberg-SCORE™ , allele database 3.5.0
Product formats vs Low Resolution Ambiguities, All samples 40%
35%
30%
25%
20%
) g(% b m a tin slu re w Lo
15%
10%
5%
0%
HLA-A
HLA-B
HLA-DRB
All samples, "classic format"
14%
16%
3,4%
All samples, new format
0,5%
1,5%
0,0%
Product formats vs Low Resolution Ambiguities, Sample Breakdown
HLA-A
HLA-B
HLA-DRB
”Classic” format
New format
”Classic” format
New format
”Classic” format
New format
All samples (n=204)
14%
0,5%
16%
1,5%
3,4%
0,0%
Random samples (n=102)
16%
1,0%
17%
1,0%
0,5%
0,0%
UCLA samples (n=102)
12%
0,0%
16%
2,0%
5,9%
0,0%
Olerup SSP® Low resolution Kits – Examples of improved low resolution HLA-A –Improved resolution for A*01,03; A*01,11 and A*03,11 samples • Resolves for example: – 01:01,03:01=03:01,03:18 and 1:01,11:01=11:01,11:94
–Improved resolution for samples containing A*26, A*66 or A*34 alleles • Resolves for example: – 01:01,26:01=03:18,26:01 – 11:01,26:03=03:75,11:10 – 26:01,66:02=26:09,26:65=26:65,43:01 – 66:01,66:02=34:01,66:12
–Improved reactivity pattern for A19-samples • A19-specific vial included – Will decrease risks of new alleles with blank reactivity patterns. Thereby sustaining resolution for homozygous samples
HLA-DR • Improved resolution for DRB1*13,13; DRB1*13,14 and DRB1*14,14 samples • Resolves for example: – – – –
13:01,14:02= 14:21,14:24 14:01,14:02=13:44/86,14:01 13:03,14:01=03:76,14:01=14xx,14xx Improved resolution for samples containing DRB1*03, DRB1*08, DRB1*07, DRB1*09 or DRB1*12
Olerup SSP® Low resolution Kits – Examples of improved low resolution HLA-B –Improved resolution for B*07,39 and B*07,40 samples • Resolves for example: – 07:02,39:01=07:69,67:01 – 07:02,39:02=48:05,67:01 – 07:02,40:01=40:xx, 40:166
–Improved resolution for B62,B63 samples. Separated from B15- and B62 serological equivalents –Exon 2-3 (Intron 2 spanning) primer pairs included
–Improved resolution for B*35 samples • With respect to B*18-alleles
• Resolves for example: – B*35:01:01:01,51:01:02=B*53:01:01,7 8:02:01
–Improved resolution for B*08:01 homozygous samples –Improved resolution through B*42 specific primer mix • Resolving various ambiguities for samples containing for example B*07, B*08, B*27 or B*39 alleles
Confidential - for internal use only
Olerup SSP® Low resolution Kits – Flexibility and Convenience • Conclusions – Low resolution kit format expansion result in a significant decrease of ambiguities for both sample populations – Possible in new format to include primer pairs spanning Class I intron 2 • resolves for example B*35:01:01:01,51:01:02 = B*53:01:01,78:02:01
• Benefits of the format change – will enable us to continue to: • cover all alleles as databases are released • Sustain high quality low resolution • Increase genotype resolution