Různé metody stanovení PCSK9 a jejich srovnání s ohledem na klinickou utilitu

Bc. Karel Snášel

Master's thesis

PCSK9 measurement and method comparison with regards to clinical utility

Abstract:

Prace predstavuje vyvoj nove ho prototypu chemiluminiscenc ní imunoanaly zy (CLIA) pro stanovení proteinu PCSK9 na automaticke m analyza toru KleeYa, ktery byl motivova n naru stají cí potr ebou efektivní ho a rychle ho stanovení PCSK9 v souvislosti s neda vno zava de nou le c bou inhibitory PCSK9. Tato le c ba pr edstavuje zleps ení moz ností terapie hypercholesterolemie a prevence kardiovaskula rní ch onemocne ní zejme na u pacientu s familia rní hypercholesterole mií a poruchami lipidove ho metabolismu. Pro uvedenou le c bu by bylo pr í nosem pr esne monitorova ní hladin PCSK9, aby byla optimalizova na jejich u c innost a minimalizova ny moz ne vedlejs í u c inky. Nove vyvinuty prototyp CLIA pro detekci PCSK9 byl navrz en tak, aby poskytoval pr esne vy sledky do 20 minut, coz je za sadní pro klinickou praxi, kde rychlost a pr esnost diagnostiky mohou vy razne ovlivnit le c ebne rozhodnutí . Nastavení me r í cí ho rozsahu prototypu vyhovuje pro vzorky se r s fyziologicky mi hodnotami i koncentracemi pro pacienty le c ene inhibitory PCSK9. Ove r ení spra vnosti nastavení prototypu CLIA PCSK9 je realizova no prostr ednictví m analyticke sady testu a klinicka spra vnost a sensitivita prostr ednictví m me r ení m rea lny ch vzorku zdravy ch da rcu a pacientu le c eny ch inhibitory PCSK9. Srovna ní s metodou ELISA proka zalo shodu se statistickou vy znamností p=0,05. Nove vyvinuty prototyp CLIA PCSK9 s automatizovany m provedení m spln uje poz adavky pro specificitu, pr esnost a rychlost analy zy a tí m pouz ití pro testova ní velke ho mnoz ství vzorku v klinicke praxi v kontextu zleps ení podmí nek pro le c bu a monitoring pacientu s familia rní hypercholesterolemií a jiny mi poruchami lipidove ho metabolismu.

Abstract:

The paper presents the development of a new prototype of chemiluminescent immunoassay (CLIA) for the determination of PCSK9 protein on the KleeYa automated analyzer, which was motivated by the increasing need for efficient and rapid determination of PCSK9 in connection with the recently introduced treatment with PCSK9 inhibitors. This treatment represents an improvement in the therapy options for hypercholesterolemia and the prevention of cardiovascular diseases, especially in patients with familial hypercholesterolemia and lipid metabolism disorders. For this treatment, precise monitoring of PCSK9 levels would be beneficial to optimize their effectiveness and minimize possible side effects. The newly developed CLIA prototype for PCSK9 detection was designed to provide accurate results within 20 minutes, which is crucial for clinical practice, where the speed and accuracy of diagnosis can significantly influence treatment decisions. The measurement range setting of the prototype is suitable for serum samples with physiological values and concentrations for patients treated with PCSK9 inhibitors. Verification of the correctness of the CLIA PCSK9 prototype settings is carried out through an analytical test panel, and clinical accuracy and sensitivity are assessed by measuring real samples from healthy donors and patients treated with PCSK9 inhibitors. Comparison with the ELISA method demonstrated agreement with statistical significance at p=0.05. The newly developed CLIA PCSK9 prototype with automated execution meets the requirements for specificity, accuracy, and speed of analysis, thus enabling its use for testing a large number of samples in clinical practice, improving conditions for the treatment and monitoring of patients with familial hypercholesterolemia and other lipid metabolism disorders.

Keywords

CLIA, ELISA, magnetické nanočástice, chemiluminiscence, proprotein konvertáza subtilisin/kexin typu 9, inhibitory PCSK9, familiární hypercholesterolemie, CLIA prototyp

Language used: Czech

Date on which the thesis was submitted / produced: 16. 5. 2024

Thesis defence

• Date of defence: 14. 6. 2024
• Supervisor: doc. Ing. Martin Mandl, CSc.
• Reader: Mgr. Jakub Sopoušek, Ph.D.

Masaryk University

Faculty of Science

Master programme / field:
Biotechnology / Biotechnology