Differentiated ANCA diagnostics using BIOCHIP Mosaics
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1 Differentiated ANCA diagnostics using BIOCHIP Mosaics Gran. MPO microdoplet Example: Antibodies against MPO and homogeneous ANA pattern
2 Anti-MPO? ANA? panca positive, anti-mpo positive gran. Final result: formalin-resistant panca, anti-mpo (Anti-PR3 ELISA: 2 RU/ml (NV < 20 RU/ml), Anti-PR3 Capture ELISA: 2 RU/ml (NV < 20 RU/ml), Anti-PR3-hn-hr ELISA: 3 RU/ml (NV < 20 RU/ml), Anti-MPO ELISA: 56 RU/ml (NV < 20 RU/ml), Anti-BPI/Elastase/ Cathepsin-G ELISA: negative, EUROLINE ANA Profile 3: negative) 2
3 canca? Anti-PR3? canca positive, anti-pr3- positive gran. Final result: canca, anti-pr3 (Anti-PR3 ELISA: > 200 RU/ml (NV < 20 RU/ml), Anti-PR3 Capture ELISA: > 200 RU/ml (NV < 20 RU/ml), Anti-PR3-hn-hr ELISA: > 200 RU/ml (NV < 20 RU/ml), Anti-MPO ELISA: 9 RU/ml (NV < 20 RU/ml), Anti-BPI ELISA: positive, Anti- Elastase/Cathepsin-G ELISA: negative, EUROLINE ANA Profile 3: negative) 3
4 Anti-MPO? ANA? Quick ANCA positive, further testing required gran. Final result: formalin-sensitive panca, anti-cathepsin G (Anti-PR3 ELISA: 2 RU/ml (NV < 20 RU/ml), Anti-PR3 Cap ture ELISA: 2 RU/ml (NV < 20 RU/ml), Anti-PR3-hn-hr ELISA: 12 RU/ml (NV < 20 RU/ml), Anti-MPO ELISA: 2 RU/ml (NV < 20 RU/ml), Anti-BPI/Ela stase ELISA: negative, Anti-Cathepsin-G ELISA: positive, EUROLINE ANA Profile 3: negative) 4
5 canca? Anti-PR3? anti-lysosomes positive gran. Final result: anti-lysosomes (Anti-PR3 ELISA: 2 RU/ml (NV < 20 RU/ml), Anti-PR3 Cap ture ELISA: 2 RU/ml (NV < 20 RU/ml), Anti-PR3-hn-hr ELISA: 3 RU/ml (NV < 20 RU/ml), Anti- MPO ELISA: 2 RU/ml (NV < 20 RU/ml), Anti-BPI/Ela stase/cathepsin-g ELISA: negative, EUROLINE ANA Profile 3: anti-dsdna borderline) 5
6 Anti-MPO? ANA? anti-nuclear membrane positive gran. Final result: anti-nuclear membrane, anti-ro-52 (Anti-PR3 ELISA: 2 RU/ml (NV < 20 RU/ ml), Anti-PR3 Capture ELISA: 2 RU/ml (NV < 20 RU/ml), Anti-PR3-hn-hr ELISA: 2 RU/ml (NV < 20 RU/ml), Anti-MPO ELISA: 2 RU/ml (NV < 20 RU/ml), Anti-BPI/Elastase/Cathepsin- G ELISA: negative, EUROLINE ANA Profile 3: anti-ro-52 positive) 6
7 Anti-MPO? ANA? Quick granular pattern, further testing required gran. Final result: anti-rnp, anti-ro-52 (Anti-PR3 ELISA: 4 RU/ml (NV < 20 RU/ml), Anti-PR3 Cap ture ELISA: 4 RU/ml (NV < 20 RU/ml), Anti-PR3-hn-hr ELISA: 9 RU/ml (NV < 20 RU/ ml), Anti-MPO ELISA: 2 RU/ml (NV < 20 RU/ml), Anti-BPI/Ela stase/cathepsin-g ELISA: negative, EUROLINE ANA Profile 3: anti-rnp positive, anti-ro-52 positive) 7
8 Anti-MPO? ANA? panca pos., anti-mpo pos., ANA-pos. gran. Final result: formalin-resistant panca, anti-mpo, ANA homogeneous, anti-ro-52 (Anti- PR3 ELISA: 2 RU/ml (NV < 20 RU/ml), Anti-PR3 -hn-hr ELISA: 2 RU/ml (NV < 20 RU/ml), Anti-MPO ELISA: 126 RU/ml (NV < 20 RU/ml), Anti-BPI/Elastase/Cathepsin-G ELISA: negative, EUROLINE ANA Profile 3: negative) 8
9 canca? Anti-PR3? Quick cytopl. pattern, further testing required gran. Final result: anti-ribosomal P proteins (Anti-PR3 ELISA: 2 RU/ml (NV < 20 RU/ml), Anti-PR3 Capture ELISA: 4 RU/ml (NV < 20 RU/ml), Anti-PR3-hn-hr ELISA: 5 RU/ml (NV < 20 RU/ml), Anti-MPO ELISA: 2 RU/ml (NV < 20 RU/ml), Anti-BPI/Elastase/Cathepsin-G ELISA: negative, EUROLINE ANA Profile 3: anti-rib. P-proteins positive) 9
10 canca? anti-centromeres positive gran. Final result: anti-centromeres (Anti-PR3 ELISA: 2 RU/ml (NV < 20 RU/ml), Anti-PR3 Capture ELISA: 2 RU/ml (NV < 20 RU/ml), Anti-PR3-hn-hr ELISA: 2 RU/ml (NV < 20 RU/ml), Anti-MPO ELISA: 2 RU/ml (NV < 20 RU/ml), Anti-BPI/Ela stase/cathepsin-g ELISA: negative, EUROLINE ANA Profile 3: anti-centromere protein B positive) 10
11 canca? ANCA, anti-pr3 and anti-mpo positive gran. Final result: ANCA, anti-pr3 and anti-mpo positive (Anti-PR3-hn-hr ELISA: 200 RU/ml (NV < 20 RU/ml), Anti-MPO ELISA: 450 RU/ml (NV < 20 RU/ml), Anti-BPI/Elastase/Cathepsin-G ELISA: negative, EUROLINE ANA Profile 3: negative) 11
12 ANCA diagnostics Pure guesswork using ethanol-fixed granulocytes alone! reliable results using BIOCHIP Mosaics! Primate liver HEp-2 cells + granulocytes FA_1200_I_UK_B04, 11/2008
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