The binding of proteinase 3 antineutrophil cytoplasmic antibodies (PR3-ANCA) varies in different ELISAs.
BACKGROUND
The demonstration of proteinase 3 specific antineutrophil cytoplasmic antibodies (PR3-ANCA), and the estimation of antibody values are useful in the diagnosis and management of patients with Wegener's granulomatosis (WG). However, external quality assessment programmes suggest that PR3-ANCA binding varies in different assays.


OBJECTIVE
To demonstrate variations in PR3-ANCA binding in different commercial and in house enzyme linked immunosorbent assays (ELISAs).


METHODS
Binding of a PR3-ANCA standard and 19 sera from patients with WG was compared in eight commercial and in house assays. Binding was expressed in different units depending on the kit.


RESULTS
One commercial assay performed unsatisfactorily. Three commercial kits produced PR3-ANCA binding (70, 102, and 84 U/ml) close to the expected value for the standard (100 U/ml). Serial dilutions of this standard were linear in only one commercial assay and the in house assay. Sera from patients with WG with borderline binding in the in house assay bound in the eight commercial kits at 0-148 kit units; low binding sera ranged from 0 to 273 units; moderately strong sera bound at 7-260 units; and strongly binding sera bound at 13-336 units. In four assays, at least one strongly positive serum bound at levels greater than the provided range.


CONCLUSIONS
Levels of antibody binding and units of binding have not been standardised in commercially available PR3-ANCA ELISAs. This may affect the diagnosis and management of patients with WG, in addition to the implementation of international guidelines for treatment.
Immunology Unit, Division of Laboratory Medicine, Austin Hospital, Heidelberg, Victoria 3084, Australia.
M
M
Trevisin
P
P
Neeson
J
J
Savige
D019268:Antibodies, Antineutrophil Cytoplasmic / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D019084:Fluorescent Antibody Technique, Indirect / Q000379:methods; D014890:Granulomatosis with Polyangiitis / Q000175:diagnosis* / Q000378:metabolism; D006801:Humans; D053146:Myeloblastin; D011933:Reagent Kits, Diagnostic / Q000592:standards; D015203:Reproducibility of Results; D012697:Serine Endopeptidases / Q000378:metabolism*
D019268:Antibodies, Antineutrophil Cytoplasmic; D011933:Reagent Kits, Diagnostic; D012697:Serine Endopeptidases; D053146:Myeloblastin