Ispitivanje preciznosti izrade diferencijalne krvne slike primjenom digitalne morfologije na uređaju Sysmex DI-60 (Cellavision) i svjetlosne mikroskopije

master's thesis

• Title (english): Determination of the precision of differential blood count preparation using digital morphology on the Sysmex DI-60 (Cellavision) analyzer and light microscopy
• Author: Elena Filipaj
• Mentor: Désirée Coen Herak (mentor)
• Mentor: Marija Miloš (mentor)
• Committee member: Desiree Coen Herak (predsjednik povjerenstva)
• Committee member: Marija Miloš (član povjerenstva)
• Committee member: Ana Kozmar (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of medical biochemistry and haematology); Zagreb
• Defense date and country: 2024-09-16, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Medical Biochemistry

Abstract

Izrada DKS-a uobičajeno se provodi primjenom triju metoda: automatiziranom metodom na hematološkim
brojačima, svjetlosnom mikroskopijom i digitalnom morfološkom analizom stanica. Izrada DKS-a na
hematološkim brojačima predstavlja prvi korak u analizi DKS-a periferne krvi. U slučaju prisutnosti patoloških
stanica i stanica s morfološkim abnormalnostima, analizator daje upozorenje o patološkom nalazu što upućuje na
potrebu izrade DKS-a svjetlosnom mikroskopijom i/ili digitalnom morfološkom analizom. Bez obzira na
tehnološki napredak, izrada DKS-a svjetlosnom mikroskopijom i dalje predstavlja zlatni standard. Međutim,
prednost primjene uređaja za digitalnu morfologiju je ušteda vremena uz visoku preciznost i točnost. Cilj ovog
diplomskog rada bio je ispitati i usporediti preciznost izrade DKS-a metodom svjetlosne mikroskopije i digitalne
morfološke analize (postupak preklasifikacije i klasifikacije) u razmazima periferne krvi sa i bez patologije,
primjenom uređaja Sysmex DI-60. Ukupno je analizirano sedam razmaza periferne krvi: tri razmaza rutinskih
uzoraka u kojima je ispitana preciznost svjetlosne mikroskopije i digitalne morfološke analize, te četiri razmaza iz
programa vanjske procjene kvalitete u kojima je ispitana preciznost digitalne morfološke analize. Temeljem
dobivenih rezultata potvrđena je hipoteza o većoj preciznosti digitalne morfološke analize u odnosu na metodu
svjetlosne mikroskopije. Iznimka su bili patološki razmazi s akutnim leukemijama u kojima se metoda svjetlosne
mikroskopije pokazala preciznijom u brojenju patoloških, nezrelih stanica periferne krvi. Nadalje, preciznost
dobivena postupkom klasifikacije na uređaju Sysmex DI-60 nije uvijek bila veća u odnosu na postupak
preklasifikacije zbog nejasno definiranih morfoloških kriterija i subjektivnosti u njihovoj procjeni. Također, u
postupku preklasifikacije na uređaju Sysmex DI-60, uočen je veći udio neidentificiranih stanica u patološkim
razmazima periferne krvi u odnosu na razmaze bez patologije, što potvrđuje nužnost primjene svjetlosne
mikroskopije u takvim slučajevima. Najmanja preciznost uočena je za nesegmentirane granulocite za sve tri
metode i za ponovljivost i za međupreciznost u razmazima sa i bez patologije.

Abstract (english)

Preparation of a differential blood count (DBC) is commonly carried out using three methods: automated analysis
on hematology analyzers, light microscopy, and digital morphological cell analysis. Performing a DBC on
hematology analyzers is the first step in the analysis of peripheral blood DBC. In the presence of pathological cells
and cells with morphological abnormalities, the analyser provides a warning indicating a pathological finding,
suggesting the need for DBC analysis using light microscopy and/or digital morphological analysis. Despite
technological advancements, DBC performed using light microscopy remains the gold standard. However, the
advantage of using digital morphology analyzers lies in time savings, along with high precision and accuracy. The
aim of this thesis was to examine and compare the precision of DBC performed by light microscopy and digital
morphological analysis (reclassification and classification processes) on peripheral blood smears with and without
pathology, using the Sysmex DI-60 analyzer. A total of seven peripheral blood smears were analyzed: three smears
of routine samples in which the precision of light microscopy and digital morphological analysis was examined,
and four smears from the external quality assessment program in which the precision of digital morphological
analysis was assessed. Based on the obtained results, the hypothesis of greater precision of digital morphological
analysis compared to the light microscopy method was confirmed. Exceptions were pathological smears with acute
leukemias, in which light microscopy proved to be more precise in counting pathological, immature peripheral
blood cells. Furthermore, the precision obtained by the classification process on the Sysmex DI-60 analyzer was
not always higher compared to the reclassification process, due to poorly defined morphological criteria and
subjectivity in their assessment. Additionally, in the reclassification process on the Sysmex DI-60 analyzer, a
higher proportion of unidentified cells was observed in pathological peripheral blood smears compared to nonpathological
smears, confirming the necessity of using light microscopy in such cases. The lowest precision was
observed for non-segmented granulocytes across all three methods, for both repeatability and intermediate
precision, in smears with and without pathology.

• Keywords: diferencijalna krvna slika; digitalna morfologija; Sysmex DI-60; svjetlosna mikroskopija; preciznost
• Keywords (english): differential blood count; digital morphology; Sysmex DI-60; light microscopy; precision
• Language: croatian
• URN:NBN: urn:nbn:hr:163:328265
• Study programme: Title: Medical biochemistry - integrated undergraduate and graduate - university study; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: sveučilišni magistar medicinske biokemije i laboratorijske medicine (sveučilišni magistar medicinske biokemije i laboratorijske medicine)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2025-01-10 13:07:40