Probir na mukopolisaharidoze iz uzorka suhe kapi mokraće

master's thesis

• Title (english): Screening for mucopolysaccharidosis from a sample of dry urine spots
• Author: Rebeka Svatić
• Mentor: Ksenija Fumić (mentor)
• Committee member: Ksenija Fumić (predsjednik povjerenstva)
• Committee member: Olga Gornik Kljaić (član povjerenstva)
• Committee member: Toma Keser (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of medical biochemistry and haematology); Zagreb
• Defense date and country: 2020-05-21, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Medical Biochemistry

Abstract

Mukopolisaharidoze su podgrupa lizosomskih bolesti nakupljanja. Ove bolesti predstavljaju izazov suvremenoj kliničkoj i laboratorijskoj dijagnostici zbog heterogenosti simptoma i raznih oblika patofiziološke manifestacije. Rano postavljanje dijagnoze povećava mogućnosti početka liječenja i oporavka, a to je moguće samo ako postoje brzi i pouzdani testovi. Nakon postavljanja kliničke sumnje, laboratorijska dijagnostika mukopolisaharidoza započinje određivanjem koncentracije glikozaminoglikana u mokraći. Inspiracija za ovaj diplomski rad dobivena je iz studije koja opisuje rezultate dvogodišnjeg FIND projekta u Španjolskoj. Jedinica za dijagnozu i liječenje kongenitalnih metaboličkih bolesti je u suradnji s MPS asocijacijom Španjolske za rijetke bolesti pokrenula FIND projekt kojim bi se rano dijagnosticirale MPS kod djece sa simptomima bolesti te podigla svijest o ovim bolestima među pedijatrima. Unutar ovog projekta razvijena je in house metoda za određivanje GAG-ova iz suhe kapi mokraće prikupljene na filter papiru. Svi postupci pripreme uzorka, reagensa i kalibracijskih točaka u ovom diplomskom radu napravljeni su po uzoru i preporuci Laboratorija za metaboličke bolesti Kliničke bolnice Santiago de Compostela, Španjolska. Cilj rada je bio ispitati in house metodu za probir mukopolisaharidoza kako bi se pojednostavio postupak slanja uzoraka u laboratorij i ubrzala dijagnostika. Dobiveni rezultati in house metodom iz uzoraka suhe kapi mokraće nisu se pokazali pouzdanima, jer se GAG-ovi neravnomjerno ekstrahiraju iz suhe kapi mokraće na filter papiru. Osim toga metoda je vremenski zahtjevnija nego što se očekivalo. Stoga ova in house metoda nije pogodna za probir na mukopolisaharidoze. Njena eventualna primjena je moguća samo iz tekućih uzorka mokraće ako laboratoriju nije dostupna druga metoda. Određivanje GAG-ova kolorimetrijskom metodom s Blyscan reagensom je pouzdana metoda probira na mukopolisaharidoze koja se koristi u većini metaboličkih laboratorija.

Abstract (english)

Mucopolysaccharidosis is a subset of lysosomal storage diseases. These diseases present a challenge to modern clinical and laboratory diagnostics due to the heterogeneity of symptoms and various forms of pathophysiological manifestation. Early diagnosis increases the chances of starting treatment and recovery, and this is only possible if there are fast and reliable tests. After setting the clinical suspicion, laboratory diagnosis of mucopolysaccharidosis begins with determining the concentration of glycosaminoglycans in the urine. The inspiration for this thesis was obtained from a study describing the results of a two-year FIND project in Spain. The Congenital Metabolic Diseases Diagnosis and Treatment Unit, in collaboration with the Spanish MPS Association for Rare Diseases, launched the FIND project to diagnose MPS early in children with symptoms and raise awareness of these diseases among pediatricians. Within this project, an in-house method was developed to determine GAG from dry spots of urine collected on filter paper. All procedures in this thesis are made according to the model and recommendation of the Laboratory for Metabolic Diseases of the Clinical Hospital Santiago de Compostela, Spain. The aim of the study was to examine the in house method for mucopolysaccharidosis screening in order to simplify the procedure of sending samples to the laboratory and speed up the diagnosis. The results obtained with in house method for samples from dry urine spots did not prove to be reliable, because GAGs are unevenly extracted from dry urine spots on filter paper. In addition, the method is more time consuming than expected. Therefore this in house method is not suitable for screening for mucopolysaccharidosis. Its possible application is possible only from liquid urine samples if no other method is available to the laboratory. Determination of GAGs by colorimetric method with Blyscan reagent is a reliable method for screening for mucopolysaccharidosis and is used in most metabolic laboratories.

• Keywords: mukopolisaharidoze; glikozaminoglikani; Blyscan test; in house metoda
• Keywords (english): mucopolysaccharidoses; glycosaminoglycans; Blyscan assay; in house method
• Language: croatian
• URN:NBN: urn:nbn:hr:163:684507
• 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: 2022-01-25 13:55:54