Title ODREĐIVANJE PROTUTIJELA NA EPSTEIN-BARROV VIRUS RAZLIČITIM IMUNOTESTOVIMA Title (english) DETECTION OF EPSTEIN-BARR VIRUS SPECIFIC ANTIBODIES BY DIFFERENT IMMUNOASSAYS Author Marijan Barišić Mentor Marina Bubonja Šonje (mentor)Committee member Maja Abram (predsjednik povjerenstva)Committee member Darinka Vučković (član povjerenstva)Committee member Marina Bubonja Šonje (član povjerenstva)Granter University of Rijeka Defense date and country 2018-09-25, Croatia Scientific / art field, BIOMEDICINE AND HEALTHCARE Abstract Epstein-Barrov virus (EBV) je jedan od najučestalijih uzročnika virusnih infekcija u ljudi. Specifična EBV-protutijela se mogu detektirati različitim serološkim postupcima u serumu zaraženih osoba metodama kao što su enzimski imunotest (EIA, engl. enzyme immunoassay), kemiluminiscentni imunotest (CLIA, engl. chemiluminescent immunoassay) i dr. Za definiranje faze infekcije najčešće se koriste testovi za određivanje IgM i IgG protutijela na antigen virusne kapside (VCA) EBV-a te IgG protutijela na EBV nuklearni antigen-1 (EBNA-1). U serumu bolesnika s akutnom primarnom infekcijom obično su prisutna IgM protutijela na VCA ili IgG i IgM protutijela na VCA, a izostaju IgG protutijela na EBNA-1. U osoba koje su preboljele infekciju prisutna su IgG protutijela na VCA uz EBNA-1 IgG, a izostaju IgM protutijela na VCA. IgM protutijela protiv VCA antigena prisutna su nekoliko mjeseci, dok su IgG protutijela protiv VCA i EBNA prisutna godinama.
Cilj istraživanja je usporediti rezultate dobivene korištenjem komercijalnih Enzygnost anti-EBV IgG i IgM testova EIA metodom na analizatoru BEP® 2000 System (Siemens, Njemačka), te Abbot Architect anti-EBV IgG i IgM testova kemiluminiscentnom metodom (CMIA, engl. chemiluminescent microparticle immuno assay) na analizatoru Architect i1000SR (Abbott, SAD) s referentnom metodom. U radu su korišteni arhivirani uzorci seruma koji su prethodno testirani u sklopu rutinske dijagnostike EBV infekcije u Kliničkom laboratoriju za kliničku mikrobiologiju KBC Rijeka. Serumi su prethodno analizirani Vircell ELISA metodom korištenjem kitova EBV VCA ELISA IgG, EBV VCA ELISA IgM i EBNA ELISA IgG (proizvođač Vircell, Granada, Španjolska), a dobiveni serološki profili potvrđeni su CLIA metodom istog proizvođača na ThunderBolt uređaju. Navedene metode su uzete kao referentne metode testiranja.
Visoka osjetljivost i specifičnost Enzygnost i Abbott komercijalnih seroloških EBV testova, te velika podudarnost rezultata dobivenih s obje korištene metode s referentnom metodom ukazuju na pouzdanost ovih komercijalnih testova u kliničkoj dijagnostičkoj primjeni. Dodatno, upotreba trećeg markera ENBA-1 IgG (Abbott Architect EBV test) omogućava bolju diskriminaciju primarne i prošle infekcije te pridonosi većoj pouzdanosti dobivenih rezultata.
Abstract (english) Epstein-Barr virus (EBV) is one of the most common causes of of viral infections in humans. Specific EBV antibodies can be detected by different serological procedures in serum of infected individuals with the help of methods such as enzyme-immunoassay (EIA), chemiluminescent immunoassay (CLIA) and others. For infection stage determination, tests for detection of IgM and IgG antibodies to EBV Viral Capsid Antigen (VCA) and IgG antibodies to Epstein-Barr Nuclear Antigen-1 (EBNA-1) are commonly used. VCA IgG and VCA IgM antibodies in the absence of EBNA-1 IgG antibodies are typically found in serum of patients with acute primary infections. Past infections are characterized by the presence of VCA IgG and EBNA-1 IgG antibodies in the absence of VCA IgM antibodies. VCA IgM antibodies persist for months, while VCA IgG and EBNA IgG antibodies persist for years.
The aim of the study is to compare results of commercially available Enzygnost Anti-EBV IgG and IgM assays by EIA method on BEP® 2000 System (Siemens, Germany), and results of Abbott Architect Anti-EBV IgG and IgM assays by chemiluminescent microparticle immunoassay (CMIA) method on Architect i1000SR analyzer (Abbott, USA), with a reference method. In this study, archived serum samples were used that were previously tested as part of routine diagnostics of EBV infections at Clinical Department for Clinical Microbiology of Clinical Hospital Centre Rijeka. The serum samples were previously tested by Vircell ELISA method using EBV VCA ELISA IgG, EBV VCA ELISA IgM and EBNA ELISA IgG kits (manufacturer Vircell, Granada, Spain), and resulting serological profiles were confirmed by CLIA method by the same manufacturer on ThunderBolt analyzer. The above methods were used as reference testing methods.
High sensitivity and specificity of commercially available Enzygnost and Abbott serological EBV assays, as well as high rate of result agreement of the two methods with the reference method indicate the reliability of these assays in clinical diganostics. Additionally, the use of a third marker, ENBA-1 IgG (Abbott Architect EBV assay), provides better discrimination of primary and past infection and contributes to better reliability of obtained results.
Keywords
EBV
CMIA
ELISA
serološke metode
Keywords (english)
EBV
CMIA
ELISA
serological methods
Language croatian URN:NBN urn:nbn:hr:184:615043 Study programme Title: Graduate university study of medical laboratory diagnostics (Biomedicine and Healthcare; clinical medical sciences) Type of resource Text File origin Born digital Access conditions Access restricted to students and staff of home institution Terms of use Created on 2018-12-13 09:47:55
