Primerjava mikroskopskega pregleda citološkega sedimenta in pretočne citometrije pri diferenciranju levkocitov v peritonealni, plevralni in sinovialni tekočini ter likvorju

Gartnar, Tanja (avtor), Jerin, Aleš (mentor), Božnar Alič, Elizabeta (komentor)

Analiza telesnih tekočin ima pomembno vlogo pri diagnostiki vnetnih, infekcijskih, malignih in drugih patoloških stanj. Poleg celokupnega števila levkocitov je ključen del laboratorijske obravnave tudi diferenciacija levkocitov, ki se tradicionalno izvaja z mikroskopskim pregledom citološkega sedimenta, vendar je ta metoda časovno zamudna in podvržena subjektivni presoji. Namen magistrske naloge je primerjati mikroskopsko metodo pregleda citološkega sedimenta z avtomatizirano metodo pretočne citometrije na analizatorju Sysmex XN-1000 z modulom za telesne tekočine ter oceniti uporabnost in zamenljivost obeh metod v klinični praksi. V nalogo je bilo vključenih 1013 vzorcev telesnih tekočin (289 vzorcev likvorja, 214 vzorcev peritonealne tekočine, 355 vzorcev plevralne tekočine in 155 vzorcev sinovialne tekočine). Rezultate smo statistično ovrednotili s Spearmanovim korelacijskim koeficientom, Passing–Bablokovo regresijo in Bland–Altmanovo analizo. Pri primerjavi ujemanja meritev med mikroskopsko in avtomatizirano metodo so rezultati pokazali dobro ujemanje za številčno koncentracijo in delež polimorfonuklearnih in mononuklearnih levkocitov. Analizator v povprečju izmeri 2,4 % več polimorfonuklearnih in 2,4 % manj mononuklearnih levkocitov, največje razlike so v peritonealni tekočini, in sicer avtomatizirana metoda izmeri 7,7 % več polimorfonuklearnih in 7,6 % manj mononuklearnih levkocitov. Primerljivost metod je pri določanju koncentracij posameznih vrst celic (nevtrofilni granulociti, limfociti, monociti) dobra. Pri limfocitih je največje odstopanje v plevralni tekočini, v kateri avtomatizirana metoda izmeri za 10,8 % več limfocitov. Metodi se nekoliko slabše ujemata pri diferenciranju monocitov, in sicer imamo negativno proporcionalno odstopanje (Passing-Bablokova regresijska enačba za delež monocitov za vse tekočine: y = 1,638 + 0,669x). Določanje eozinofilnih granulocitov z avtomatizirano metodo je težavno, saj so prisotni v nizkih koncentracijah in imamo veliko relativno odstopanje med metodama. Ugotovljene razlike med mikroskopsko in avtomatizirano metodo so bistveno manjše od diagnostičnih mejnih vrednosti za ključna klinična stanja, zato ni verjetno, da bi te razlike vodile do napačne diagnoze spontanega bakterijskega peritonitisa, meningitisa, pljučnega empiema ali septičnega artritisa.

• Jezik: Slovenski jezik
• Ključne besede: telesne tekočine, diferenciacija levkocitov, citološki sediment, pretočna citometrija, Sysmex XN-1000
• Vrsta gradiva: Magistrsko delo/naloga
• Organizacija: FFA - Fakulteta za farmacijo
• Leto izida: 2026
• PID: 20.500.12556/RUL-181706
• Datum objave v RUL: 14.04.2026

Sekundarni jezik

• Jezik: Angleški jezik
• Naslov: Comparative evaluation of manual differential count and flow cytometry in peritoneal, pleural, synovial, and cerebrospinal fluid

Izvleček

The analysis of body fluids plays an important role in the diagnosis of inflammatory, infectious, malignant, and other pathological conditions. In addition to the total leukocyte count, a key component of laboratory evaluation is leukocyte differentiation, which is traditionally performed by microscopic examination of cytological slide. However, this method is time-consuming and subjective. The aim of this master’s thesis is to compare the microscopic examination of cytological slide with automated flow cytometric method using the Sysmex XN-1000 analyzer with body fluid module for leukocyte differentiation, and to evaluate the use and interchangeability of both methods in clinical practice. A total of 1013 body fluid samples were included in the study (289 cerebrospinal fluid, 214 peritoneal fluid, 355 pleural fluid, and 155 synovial fluid samples). The results were statistically evaluated using Spearman’s coefficient, Passing–Bablok regression, and Bland–Altman plot. A good agreement between methods was found for both absolute and relative polymorphonuclear and mononuclear counts. On average, the analyzer measured 2,4% more polymorphonuclear leukocytes and 2,4% fewer mononuclear leukocytes, with the largest differences observed in peritoneal fluid, where the automated method measured 7,7% more polymorphonuclear leukocytes and 7,6% fewer mononuclear leukocytes. The comparability of the methods for measuring individual cell populations (neutrophils, lymphocytes, and monocytes) was generally good. The greatest differences for lymphocytes were observed in pleural fluid, where the automated method measured 10,8% more lymphocytes. The agreement between methods was poorer for monocyte differentiation, showing a negative proportional bias (Passing–Bablok regression equation for relative monocyte count across all fluids: y = 1,638 + 0,669x). Measuring eosinophils using the automated method was challenging due to their low concentrations and the large relative deviations between methods. The observed differences between the microscopic and automated method were substantially smaller than the diagnostic decision limits for key clinical conditions. Therefore, it is unlikely that these differences would lead to misdiagnosis of spontaneous bacterial peritonitis, meningitis, pleural empyema, or septic arthritis.

• Ključne besede: body fluid, leukocyte differential count, cytospin slide, flow cytometry, Sysmex XN-1000