Verification, implementation and harmonization of automated chemiluminescent immunoassays for MPO- and PR3-ANCA detection

Ogrič, Manca (author), Švec, Tinka (author), Mrak Poljšak, Katjuša (author), Žigon, Polona (author), Hočevar, Alojzija (author), Čučnik, Saša (author)

Objectives: Antineutrophil cytoplasmic antibody (ANCA) testing assists clinicians diagnose ANCA-associated vasculitis (AAV). We aimed to verify and harmonize chemiluminescent immunoassays for the detection of myeloperoxidase (MPO)- and proteinase 3 (PR3)-ANCA. Methods: An in-house ELISA, a capture ELISA, and a chemiluminescent assay QUANTA Flash on a BIO-FLASH analyzer were used to detect MPO- and PR3-ANCA in sera from 39 patients with AAV, 55 patients with various non-AAV, and 66 patients with connective tissue diseases. The results of the assays were evaluated, and their clinical performance was assessed. The precision and linearity of the QUANTA Flash assays were determined, and likelihood ratios (LRs) for AAV at diagnosis were calculated. Results: The precision and linearity of the QUANTA Flash assays were confirmed. Overall agreement between 97.5 and 98.8 % and Cohen’s kappa coefficients between 0.861 and 0.947 were observed for the results of the QUANTA Flash assays and ELISAs. The diagnostic sensitivity, specificity, and ROC analysis of the assays for AAV were statistically similar (in-house ELISA 89.7 %, 95.0 %, and 0.937; capture ELISA 92.3 %, 98.3 %, and 0.939; and QUANTA Flash 89.7 %, 95.9 %, and 0.972). For the QUANTA Flash assay results, the interval-specific LRs for AAV at diagnosis were: 0–8 CU had LR 0.08, 8–29 CU had LR 1.03, 29–121 CU had LR 7.76, 121–191 CU had LR 12.4, and >191 CU had LR ∞. Conclusions: The QUANTA Flash MPO and PR3 assays provide precise and consistent results and have comparable clinical utility for AAV. The calculated LRs were consistent with published LRs, confirming the utility of LRs for harmonization of ANCA results.

• Language: English
• Keywords: antineutrophil cytoplasmic antibodies, ANCA, proteinase 3, PR3, myeloperoxidase, MPO, ANCA-associated vasculitis, AAV, chemiluminescent immunoassay, likelihood ratios, LRs
• Work type: Article
• Typology: 1.01 - Original Scientific Article
• Organization: MF - Faculty of Medicine; FFA - Faculty of Pharmacy
• Publication status: Published
• Publication version: Version of Record
• Year: 2024
• Number of pages: Str. 682–689
• Numbering: Vol. 62, iss. 4
• PID: 20.500.12556/RUL-154819
• UDC: 616-097
• ISSN on article: 1437-4331
• DOI: 10.1515/cclm-2023-0764
• COBISS.SI-ID: 169691907
• Publication date in RUL: 04.03.2024

Record is a part of a journal

• Title: Clinical chemistry and laboratory medicine
• Shortened title: Clin. chem. lab. med.
• Publisher: De Gruyter, European Federation of Clinical Chemistry and Laboratory Medicine
• ISSN: 1437-4331
• COBISS.SI-ID: 680355

Licences

• License: CC BY 4.0, Creative Commons Attribution 4.0 International
• Link: http://creativecommons.org/licenses/by/4.0/
• Description: This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.

Secondary language

• Language: Slovenian
• Keywords: antinevtrofilna citoplazemska protitelesa, ANCA, proteinaza 3, PR3, mieloperoksidaza, MPO, vaskulitis, povezan z ANCA, AAV, kemiluminiscentni imunski test, razmerja verjetnosti, LR

Projects

• Funder: ARRS - Slovenian Research Agency