- Legally Marketed Device
Biotest Diagnostics claims substantial equivalence to the DRB-SSO Typing Kit (BK950015) currently in commercial distribution by Biotest Diagnostics.
- Device Description
The Biotest DRB-SSO Typing Kit is a highly sensitive dot blot assay. In the first step, genomic DNA is purified and the relevant genetic region - for the DRB-SSO Typing Kit this is a DRB fragment - is replicated using PCR *.
Two PCR amplifications with primer pairs DRB-G5/DRB-86G and DRB-G5/DRB-86T, respectively, designed towards the glycine/valine dimorphism at codon 86 of DRB sequences are performed in the high resolution assay. The two primer pairs amplify two non-overlapping groups of DRB sequences; primer pair DRB-G5/DRB-86G amplifies DRB alleles encoding amino acid residue glycine at position -86-, primer pair DRB-G5/DRB-86T amplifies those DRB alleles encoding amino acid residue -86- valine.
The DNA from the two PCR products is attached to a membrane, denatured and, at the same time, immobilized. If the DNA in the sample contains complementary sections, HLA allele-specific oligonucleotide probes can hybridize to the single-stranded DNA (oligotyping, SSO). Unbound oligonucleotide probes are removed by washing. The oligonucleotide probes are labeled with diaoxigenin **.
The presence of the DNA-oligonucleotide complex is detected by means of a specific enzyme-labeled antibody (anti-digoxigenin alkaline phosphatase conjugate). The presence of the bound antibody is demonstrated in a subsequent enzyme reaction, in which a color product is formed. All reactions, including the color reaction, are carried out on the membrane bottom of a microplate well. After completion of the test, the membrane is stripped off, evaluated and archived.
- Intended Use
The Biotest High Resolution Supplementary Kit is intended for high resolution HLA-DRB typing using the DRB-SSO Typing Kit, an oligonucleotide probe detection system.
- Comparison with Predicate Device
A summary comparison of the features of the Biotest High Resolution Supplementary Kit and the DRB-SSO Typing Kit is provided in Table 1 below:
Table 1
Feature Comparison of High Resolution Supplementary Kit and DRB-SSO Typing Kit
| |
High Resolution |
DRB-SSO |
| Intended Use |
HLA-DRB Typing |
HLA-DRB Typing |
| Assay Method |
Oligonucleotide Probe Detection |
Oligonucleotide Probe Detection |
| Reactive Ingredients |
Digoxigenin labeled SSO Probes Primer |
Digoxigenin labeled SSO Probes Primer |
| Specimen: |
|
|
| Type |
Anticoagulated blood
(EDTA, citrate, heparin) |
Anticoagulated blood
(EDTA, citrate, heparin |
| Min. Volume |
500ml x 2 |
500ml
|
| Results: |
|
| Evaluation |
Visual color |
Visual Color |
| Interpretation |
Pattern of positive reactions
(-86G-group)
(computer or manual scheme) |
Pattern of positive reactions
(computer or manual scheme) |
| Kit Size |
15 tests |
15 tests |
- Performance Data
The Biotest High Resolution DRB-SSO Typing Kit was compared to commercially available anti-HLA serological test by testing 410 specimens (well characterized, serologically known HLA panel specimens). There was 99.5% concordance (408/410) between the DRB-SSO Typing results and the results obtained using the anti-HLA serological tests.
Upon further testing to resolve discordants using additional methods (standardized in-house DNA methods), there was 100% concordance (410/410).
