master's thesis
Verification of reagent for determining the concentration of human chorionic gonadotropin by electro-chemiluminescence immunoassay method

Monika Doželenčić (2015)
Sveučilište u Zagrebu
Farmaceutsko-biokemijski fakultet
Zavod za medicinsku biokemiju i hematologiju
Metadata
TitleVerifikacija reagensa za određivanje koncentracije humanog korionskog gonadotropina metodom imunotesta elektrokemiluminiscencijom
AuthorMonika Doželenčić
Mentor(s)Ana-Maria Šimundić (thesis advisor)
Abstract
Cilj ovog rada bio je ispitati izvedbene značajke metode za određivanje koncentracije humanog korionskog gonadotropina u serumu, na Roche cobas e 411 analizatoru, odnosno ispitati nepreciznost, usporedivost metoda, utjecaj prijenosa analita („carry over“) i utjecaj hemolize na određivanje hCG-a u serumu. Ispitivana je ponovljivost i međupreciznost, izračunom koeficijenata varijacije (KV, %) u uzorcima komercijalnih kontrola na dvije koncentracijske razine (PCU 1 i PCU 2), tijekom pet dana u triplikatu. Kriteriji prihvatljivosti koji su korišteni su kriteriji zadani od proizvođača: KVP(PCU 1)=2,1 %; KVP(PCU 2)=4,0 %; KVM(PCU 1)=2,6 % i KVM(PCU 2)=4,6 %. Usporedivost je ispitivana određivanjem koncentracije hCG-a u serumu u 40 uzoraka zadanog koncentracijskog raspona (0,6-10000 mIU/mL), u endokrinološkom laboratoriju i KZZK-u, u KBC-u Sestre Milosrdnice. Korišteni kriteriji prihvatljivosti su prema RCPA. Utjecaj prijenosa analita („carry over“) ispitan je analiziranjem uzorka visoke koncentracije (>10000 mIU/mL) u duplikatu i odmah potom uzorka niske koncentracije (< 0,1 mIU/mL) u triplikatu, prema IUPAC protokolu. Ispitivanje utjecaja hemolize rađeno je prema CLSI EP7-A2 protokolu. Uzorci su pripremljeni serijskom dilucijom poola seruma sa hemolizatom plazme koji je dobiven metodom osmotskog šoka, sa tri koncentracijske razine hCG-a. Svaka koncentracijska razina sadržavala je rastuće koncentracije hemoglobina (0; 2,5; 5,0; 7,5 i 10,0 g/L). Korišteni kriteriji su prema RCPA. Analize su izvršene na Roche cobas e 411 analizatoru. Za statistička obradu koristili su se Microsoft Excel i MedCalc programi. Međupreciznost i ponovljivost za PCU 2 zadovoljili su kriterije, dok za PCU 1 međupreciznost je bila unutar zadanih kriterija, a ponovljivost nije. Rezultati usporedivosti pokazali su da je odstupanje unutar kriterija kroz cijeli koncetracijski raspon. Utjecaja prijenosa analita („carry over“) iznosi 0 %. Rezultati utjecaja hemolize su unutar kriterija, osim za nisku koncentracijsku razinu (7,9 mIU/mL) pri koncentracijama hemoglobina > 5 g/L odstupanje je izvan kriterija. Možemo zaključiti da je nepreciznost zadovoljavajuća. Metode su potpuno usporedive. Rezultati ne pokazuju utjecaj prijenosa analita („carry over“) na određivanje koncentracije hCG-a u serumu. Hemoliza utječe na niskoj koncentracijskoj razini iznad 5 g/L hemoglobina u serumu. Postupak verifikacije je zadovoljio kriterije i analizat se može pustiti u rutinski rad.
Keywordshuman chorionic gonadotropin hCG verification hemolysis carry over Roche cobas e 411
Parallel title (English)Verification of reagent for determining the concentration of human chorionic gonadotropin by electro-chemiluminescence immunoassay method
Committee MembersAna-Maria Šimundić
Marija Grdić Rajković
Olga Gornik
GranterSveučilište u Zagrebu
Farmaceutsko-biokemijski fakultet
Lower level organizational unitsZavod za medicinsku biokemiju i hematologiju
PlaceZagreb
StateCroatia
Scientific field, discipline, subdisciplineBIOMEDICINE AND HEALTHCARE
Pharmacy
Medical Biochemistry
Study programme typeuniversity
Study levelintegrated undergraduate and graduate
Study programmeMedical Biochemistry
Academic title abbreviationmag. med. biochem.
Genremaster's thesis
Language Croatian
Defense date2015-08-31
Parallel abstract (English)
The aim of this study, was to examine the performance specifications of the method for determining concentration of human chorionic gonadotropin in serum, on Roche cobas e 411 analyzer, i.e. to examine imprecision, comparability, the impact of carry over and the impact of hemolysis. Repeatability and imprecision were examined by calculating the coefficient of variation (CV, %) in samples of commercial controls in two concentration levels (PCU 1 and PCU 2) during five days in triplicate. Performance criteria declared by the manufacturer were: CVP(PCU 1)=2,1 %; CVP(PCU 2)=4,0 %; CVM(PCU 1)=2,6 % and CVM(PCU 2)=4,6 %. Comparability of methods was examined determining the concentration of hCG in serum in 40 samples within given concentration range (0,6-10000 mIU/mL), in endocrine laboratory and in KZZK, KBC Sestre Milosrdnice. The RCPA acceptability criteria were used. The impact of carry over was examined by analyzing the sample with high concetrations of analyte (> 10000 mIU/mL) in duplicate and immediately after a sample with low concentrations of analyte (< 0,1 mIU/mL) in triplicate, using the IUPAC protocol. The impact of hemolysis was examined using the CLSI EP7-A2 protocol. Samples were prepared by serial dilution of pool serum spiked with plasma hemolysate obtained by osmotic shock, in three concentrations levels of hCG. Each level contained increasing concentrations of hemoglobin (0; 2,5; 5,0; 7,5 and 10,0 g/L). The RCPA acceptability criteria were used. All analysis were performed on Roche cobas e 411 analyzer. Statistical analysis was performed using Microsoft Excel and MedCalc programs. Imprecision and repeatability for PCU 2, and imprecision for PCU 1 were within the given criteria. For compabarability, results showed bias within the allowable criteria trough entire concentration range. The impact of carry over was 0 %. Results of the effect of hemolysis were within the acceptance criteria, with the exception of the lowest concentration level (7,9 mIU/mL), where hemolysis shows significant interference in samples with hemoglobin concentrations > 5 g/L. We conclude that method imprecision is satisfactory. Methods are completely comparable. Results show no impact of carry over. Hemolysis has effect at lower concentrations of analyte with hemoglobin oncetrations of hemoglobin > 5 g/L. The verification procedure was satisfying and analyte can be used in routine laboratory work.
Parallel keywords (Croatian)humani korionski gonadotropin hCG verifikacija hemoliza prijenos analita Roche cobas e 411
Resource typetext
Access conditionAccess restricted to students and staff of home institution
Terms of usehttp://rightsstatements.org/vocab/InC/1.0/
URN:NBNhttps://urn.nsk.hr/urn:nbn:hr:163:110950
CommitterPetra Gašparac