Određivanje anti-AQP4 i anti-MOG antitijela u diferencijalnoj dijagnostici demijelinizirajućih bolesti SŽS-a

master's thesis

• Title (english): Determination of anti-AQP4 and anti-MOG antibodies in differential diagnosis of demyelinating diseases
• Author: Helena Kanižić
• Mentor: Željka Vogrinc (mentor)
• Committee member: Željka Vogrinc (predsjednik povjerenstva)
• Committee member: Dunja Rogić (član povjerenstva)
• Committee member: Ksenija Fumić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of medical biochemistry and haematology); Zagreb
• Defense date and country: 2022-08-04, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Medical Biochemistry

Abstract

U autoimune, kronične demijelinizirajuće bolesti SŽS-a opisane u ovom radu spadaju multipla
skleroza (MS), poremećaji iz spektra optičkog neuromijelitisa (NMOSD) te bolesti vezane uz antitijela na
MOG (MOGAD). Diferencijalnu dijagnostiku ovih bolesti olakšalo je otkriće visokospecifičnih antitijela
pronađenih u NMOSD pacijenata, anti-AQP4 antitijela te visokospecifičnih antitijela pronađenih u
MOGAD pacijenata, anti-MOG antitijela. Rano postavljanje dijagnoze je ključno za pravilan odabir
terapije, a određivanjem antitijela na AQP4 i MOG može se znatno ubrzati i olakšati postavljanje
dijagnoze. Laboratorijski testovi koji se koriste za dokazivanje antitijela moraju zadovoljiti visoke kriterije
osjetljivosti, specifičnosti i točnosti da bi se mogli primjenjivati u rutinskom radu. Od mnogih metoda koje
su opisane za određivanje navedenih antitijela ističe se metoda indirektne imunofluorescencije temeljene
na stanicama (CBA IIF) transficiranim antigenima AQP4 i MOG. U radu su uspoređene dvije metode za
dokazivanje antitijela na AQP4, ranije korištena ELISA metoda i nova metoda CBA IIF te je ispitana
njihova dijagnostička osjetljivost, specifičnost i točnost u stvarnim laboratorijskim uvjetima. CBA IIF test
za kombinirano određivanje antitijela na AQP4 i MOG proizvođača EUROIMMUN pokazao se kao
visoko osjetljiv i visoko specifičan te jednostavan za izvođenje uz minimalno utrošeno vrijeme i resurse.
Rezultati ovog rada su potvrdili veću osjetljivost ELISA testa za određivanje antitijela na AQP4 u odnosu
na CBA IIF test (92% naprema 84%) dok je CBA IIF test specifičniji (94% u odnosu na 82%). Točnost
testa je veća kod CBA IIF testa (90,67%) u odnosu na ELISA test (85,33%). Zbog samo jednog pozitivnog
nalaza antitijela na MOG nije bilo moguće prikazati i interpretirati rezultate osjetljivosti i specifičnosti
testa jer je broj ispitanih uzoraka bio premalen. Istraživanje bi trebalo proširiti na veći broj ispitanika te
obuhvatiti i zdrave osobe kako bi se dobili potpuno vjerodostojni podaci za našu populaciju.

Abstract (english)

The autoimmune, chronic demyelinating diseases of the CNS described in this thesis include
multiple sclerosis (MS), neuromyelitis optica spectrum disorders (NMOSD), and myelin oligodendrocyte
glycoprotein antibody-associated disease (MOGAD). Differential diagnosis of these diseases was
facilitated by the discovery of highly specific antibodies found in NMOSD patients, anti-AQP4 antibodies,
and highly specific antibodies found in MOGAD patients, anti-MOG antibodies. Early diagnosis is key for
the selection of the right therapy and determination of antibodies to AQP4 and MOG can bring to a
significantly faster and much easier diagnosis.. Laboratory tests used to determine antibodies must meet
high criteria of sensitivity, specificity, and accuracy to be used in the routine. Of the many methods
described for determining these antibodies, the cell-based indirect immunofluorescence (CBA IIF) with
cells transfected with AQP4 and MOG stands out. In this thesis two methods for the determination of anti-
AQP4 antibodies, the previously used ELISA method and the new CBA IIF method are compared, and
their diagnostic sensitivity, specificity, and accuracy in real laboratory conditions are also examined. The
CBA IIF test for the combined determination of antibodies to AQP4 and MOG made by EUROIMMUN
has proved to be highly sensitive, highly specific, and easy to perform with minimal time and resources
spent. The results of this research confirmed a higher sensitivity of the ELISA test for determining
antibodies to AQP4 compared to the CBA IIF test (92% to 84%) while the CBA IIF test is more specific
(94% vs. 82%). The CBA IIF test has higher accuracy (90.67%) than the ELISA test (85.33%). Due to only
one positive result of anti-MOG antibodies, it was not possible to present and interpret the results of the
sensitivity and specificity of the test because of the too small number of examined samples. The research
should be extended to a larger number of respondents and include healthy controls to obtain fully credible
data for our population.

• Keywords: MOG; AQP-4; MS; NMOSD; MOGAD; EUROIMMUN; indirektna imunofluorescencija
• Keywords (english): MOG; AQP-4; MS; NMOSD; MOGAD; EUROIMMUN; indirect immunofluorescence
• Language: croatian
• URN:NBN: urn:nbn:hr:163:991380
• Study programme: Title: Medical biochemistry; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: magistar/magistra medicinske biokemije i laboratorijske medicine (magistar/magistra medicinske biokemije i laboratorijske medicine)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2022-09-09 14:20:47