Evaluation of capture ELISA for detection of antineutrophil cytoplasmic antibodies directed against proteinase 3 in Wegener's granulomatosis: First results from a multicentre study

Elena Csernok, Julia Holle, Bernhard Hellmich, Jan Willem, Cohen Tervaert, Cees G M Kallenberg, Peter C. Limburg, John Niles, Gouli Pan, Ulrich Specks, Kerstin Westman, Jörgen Wieslander, Kirsten De Groot, Wolfgang L. Gross

Research output: Contribution to journalArticle

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Abstract

Objective: To evaluate the performance characteristics of direct and capture ELISA for the detection of PR3-ANCA in Wegener's granulomatosis (WG) in international ANCA reference laboratories. Methods: Serum samples were derived from patients with histological and clinical diagnosis of WG (n=60), rheumatoid arthritis (RA) (n=30) and healthy controls (n=30). Each of them was tested for the presence of ANCA by indirect immunofluorescence technique (IFT), direct and capture ELISA in six international reference laboratories (Massachusetts General Hospital, Boston; Wieslab AB, Lund; University of Maastricht; University Hospital Groningen; Mayo Clinic, Rochester; Rheumaklinik Bad Bramstedt/University of Schleswig-Holstein Campus Lübeck). Each centre tested the sera according to their house protocols of IFT and ELISA. The diagnostic performance of each test was estimated by receiver operating characteristic curve analysis and sensitivity and specificity in detection of ANCA/PR3-ANCA were calculated for the respective methods. Results: In patients histologically and clinically known as WG, the detection of ANCA by IFT varied between 52 and 83% among the participating centres. PR3-ANCA positivity with the different ELISAs ranged from 53 to 80% in direct ELISA and from 72 to 76% in capture ELISA. While most capture ELISAs successfully detected PR3-ANCA, there were significant differences between IFT and direct ELISA results between laboratories. ROC curve analysis demonstrated that in five of six laboratories the overall diagnostic performance of capture ELISA was superior to IFT and direct ELISA, respectively. Conclusion: Capture ELISA is a highly sensitive assay for detection of PR3-ANCA in WG and should be used in conjunction with compatible clinical picture and histological evidence.

Original languageEnglish (US)
Pages (from-to)174-180
Number of pages7
JournalRheumatology
Volume43
Issue number2
DOIs
StatePublished - Feb 2004

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Myeloblastin
Antineutrophil Cytoplasmic Antibodies
Granulomatosis with Polyangiitis
Multicenter Studies
Enzyme-Linked Immunosorbent Assay
ROC Curve
Direct Fluorescent Antibody Technique
Fluorescent Antibody Technique
Indirect Fluorescent Antibody Technique
Serum
General Hospitals
Rheumatoid Arthritis

Keywords

  • ANCA
  • ELISA
  • IFT
  • Proteinase 3
  • Wegener's granulomatosis

ASJC Scopus subject areas

  • Neuroscience(all)
  • Rheumatology

Cite this

Evaluation of capture ELISA for detection of antineutrophil cytoplasmic antibodies directed against proteinase 3 in Wegener's granulomatosis : First results from a multicentre study. / Csernok, Elena; Holle, Julia; Hellmich, Bernhard; Willem, Jan; Tervaert, Cohen; Kallenberg, Cees G M; Limburg, Peter C.; Niles, John; Pan, Gouli; Specks, Ulrich; Westman, Kerstin; Wieslander, Jörgen; De Groot, Kirsten; Gross, Wolfgang L.

In: Rheumatology, Vol. 43, No. 2, 02.2004, p. 174-180.

Research output: Contribution to journalArticle

Csernok, E, Holle, J, Hellmich, B, Willem, J, Tervaert, C, Kallenberg, CGM, Limburg, PC, Niles, J, Pan, G, Specks, U, Westman, K, Wieslander, J, De Groot, K & Gross, WL 2004, 'Evaluation of capture ELISA for detection of antineutrophil cytoplasmic antibodies directed against proteinase 3 in Wegener's granulomatosis: First results from a multicentre study', Rheumatology, vol. 43, no. 2, pp. 174-180. https://doi.org/10.1093/rheumatology/keh028
Csernok, Elena ; Holle, Julia ; Hellmich, Bernhard ; Willem, Jan ; Tervaert, Cohen ; Kallenberg, Cees G M ; Limburg, Peter C. ; Niles, John ; Pan, Gouli ; Specks, Ulrich ; Westman, Kerstin ; Wieslander, Jörgen ; De Groot, Kirsten ; Gross, Wolfgang L. / Evaluation of capture ELISA for detection of antineutrophil cytoplasmic antibodies directed against proteinase 3 in Wegener's granulomatosis : First results from a multicentre study. In: Rheumatology. 2004 ; Vol. 43, No. 2. pp. 174-180.
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abstract = "Objective: To evaluate the performance characteristics of direct and capture ELISA for the detection of PR3-ANCA in Wegener's granulomatosis (WG) in international ANCA reference laboratories. Methods: Serum samples were derived from patients with histological and clinical diagnosis of WG (n=60), rheumatoid arthritis (RA) (n=30) and healthy controls (n=30). Each of them was tested for the presence of ANCA by indirect immunofluorescence technique (IFT), direct and capture ELISA in six international reference laboratories (Massachusetts General Hospital, Boston; Wieslab AB, Lund; University of Maastricht; University Hospital Groningen; Mayo Clinic, Rochester; Rheumaklinik Bad Bramstedt/University of Schleswig-Holstein Campus L{\"u}beck). Each centre tested the sera according to their house protocols of IFT and ELISA. The diagnostic performance of each test was estimated by receiver operating characteristic curve analysis and sensitivity and specificity in detection of ANCA/PR3-ANCA were calculated for the respective methods. Results: In patients histologically and clinically known as WG, the detection of ANCA by IFT varied between 52 and 83{\%} among the participating centres. PR3-ANCA positivity with the different ELISAs ranged from 53 to 80{\%} in direct ELISA and from 72 to 76{\%} in capture ELISA. While most capture ELISAs successfully detected PR3-ANCA, there were significant differences between IFT and direct ELISA results between laboratories. ROC curve analysis demonstrated that in five of six laboratories the overall diagnostic performance of capture ELISA was superior to IFT and direct ELISA, respectively. Conclusion: Capture ELISA is a highly sensitive assay for detection of PR3-ANCA in WG and should be used in conjunction with compatible clinical picture and histological evidence.",
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AU - Willem, Jan

AU - Tervaert, Cohen

AU - Kallenberg, Cees G M

AU - Limburg, Peter C.

AU - Niles, John

AU - Pan, Gouli

AU - Specks, Ulrich

AU - Westman, Kerstin

AU - Wieslander, Jörgen

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AU - Gross, Wolfgang L.

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N2 - Objective: To evaluate the performance characteristics of direct and capture ELISA for the detection of PR3-ANCA in Wegener's granulomatosis (WG) in international ANCA reference laboratories. Methods: Serum samples were derived from patients with histological and clinical diagnosis of WG (n=60), rheumatoid arthritis (RA) (n=30) and healthy controls (n=30). Each of them was tested for the presence of ANCA by indirect immunofluorescence technique (IFT), direct and capture ELISA in six international reference laboratories (Massachusetts General Hospital, Boston; Wieslab AB, Lund; University of Maastricht; University Hospital Groningen; Mayo Clinic, Rochester; Rheumaklinik Bad Bramstedt/University of Schleswig-Holstein Campus Lübeck). Each centre tested the sera according to their house protocols of IFT and ELISA. The diagnostic performance of each test was estimated by receiver operating characteristic curve analysis and sensitivity and specificity in detection of ANCA/PR3-ANCA were calculated for the respective methods. Results: In patients histologically and clinically known as WG, the detection of ANCA by IFT varied between 52 and 83% among the participating centres. PR3-ANCA positivity with the different ELISAs ranged from 53 to 80% in direct ELISA and from 72 to 76% in capture ELISA. While most capture ELISAs successfully detected PR3-ANCA, there were significant differences between IFT and direct ELISA results between laboratories. ROC curve analysis demonstrated that in five of six laboratories the overall diagnostic performance of capture ELISA was superior to IFT and direct ELISA, respectively. Conclusion: Capture ELISA is a highly sensitive assay for detection of PR3-ANCA in WG and should be used in conjunction with compatible clinical picture and histological evidence.

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KW - Wegener's granulomatosis

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