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Updated: Aug 1, 2026
Determining the Phagocytic Activity of Clinical Antibody Samples
Published on: November 30, 2011
Relationship between anti-neutrophil cytoplasmic antibody determined with conventional binding and the capture assay,
K Gisslén1, J Wieslander, G Westberg
1Department of Nephrology, Sahlgrenska University Hospital, Göteborg, Sweden.
Insights
The capture ELISA method for detecting anti-neutrophil cytoplasmic antibodies (ANCA) is more sensitive for diagnosing vasculitis relapses than direct ELISA. However, its specificity is lower, as high ANCA levels can occur without active disease.
Area of Science:
- Immunology
- Nephrology
- Rheumatology
Background:
- Anti-neutrophil cytoplasmic antibody (ANCA) detection is crucial for diagnosing and monitoring vasculitis.
- Different assay methods for ANCA may yield varying results, impacting clinical interpretation.
Purpose of the Study:
- To compare the efficacy of two different anti-neutrophil cytoplasmic antibody (ANCA) detection methods in predicting the long-term clinical course of vasculitis.
- To evaluate the relationship between PR3-ANCA levels measured by capture ELISA and direct ELISA with vasculitis relapses.
Main Methods:
- Retrospective analysis of 245 anti-neutrophil cytoplasmic antibody (ANCA) determinations from 10 patients with Wegener's granulomatosis or microscopic polyarteritis.
- Comparison of two assays for proteinase 3-ANCA (PR3-ANCA): conventional direct binding ELISA and capture ELISA using monoclonal antibodies.
- Long-term follow-up (mean 9 years) with frequent blood sample collection for ANCA analysis.
Main Results:
- The capture ELISA detected 100% of active vasculitis episodes, compared to 79% by conventional direct ELISA.
- In all cases of clinical relapse, the capture ELISA showed increasing or very high anti-neutrophil cytoplasmic antibody (ANCA) titres.
- A correlation between capture ANCA levels and clinical course was observed in 7 out of 10 patients.
Conclusions:
- Capture ELISA for PR3-ANCA demonstrates higher sensitivity in detecting vasculitis relapses compared to direct ELISA.
- The specificity of capture ELISA for anti-neutrophil cytoplasmic antibody (ANCA) was limited, as elevated levels were observed in patients without active disease.
Objectives:
To evaluate the relationship between anti-neutrophil cytoplasmic antibody (ANCA) measured with two different methods and long-term clinical course in vasculitis.
Design:
Retrospective determination of ANCA with two different assays for detection of PR3-ANCA, conventional direct binding ELISA and capture ELISA using monoclonal antibodies against PR3. The 245 ANCA determinations were performed from frozen blood samples collected three to four times a year in each patient.
Setting:
Department of Nephrology at a Swedish University Hospital.
Subjects:
A total of 10 ANCA-positive patients with vasculitis caused by Wegener's granulomatosis (WG) or microscopic polyarteritis (MPA) and a very long follow-up time (mean 9 years, range 5-15.5 years).
Results:
The total number of episodes with active vasculitis was 29 and all of them (100%) were detected by the capture technique whilst the conventional technique detected 23 (79%). The mean number of episodes with active disease requiring treatment with steroids and cytotoxic drugs was three per patient (range 1-6). At the time of clinical relapse of the vasculitis disease, the ANCA titre using the capture technique was either increasing or showed a very high value in all cases. The pattern of capture ANCA response could be subdivided into three categories: a close (four patients), an intermediate (three patients), and no (three patients) relationship between capture ANCA level and long-term clinical course.
Conclusion:
Detection of PR3-ANCA by the capture ELISA showed a higher sensitivity than that obtained by the direct ELISA in diagnosing relapse during follow-up of patients with vasculitis. The specificity of the capture ANCA was, however, low, as high levels occurred in patients without clinical disease activity.
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