Author/Editor		Vučković, Joško; Pavlič, Rok; Grat, Mateja; Žuran, Ivan; Krivec, Štefka
Title		Image DNA analysis of bone marrow megakaryocytes in differential diagnosis of thrombocytosis
Translated title		Slikovna analiza DNK megakariocitov kostnega mozga v diferencialni diagnozi trombocitoze
Type		članek
Source		Zdrav Vestn
Vol. and No.		Letnik 73, št. Suppl 1
Publication year		2004
Volume		str. I-23-5
Language		eng
Abstract		Background. Megakaryocyte (Mk) nuclear DNA content was measured by image analysis using new S-Form software. Feulgen-stained bone marrow aspirates from 11 patients with reactive thrombocytosis and 22 patients with clonal thrombocytosis of chronic myeloproliferative disorders (CMPDs) were analysed. Ploidy of 200 Mk was calculated for each patient and the fractions of hyperploid cells were compared. The presence of more than 5% of Mk with nuclear ploidy > 96N or >= 2% of Mk with nuclear ploidy > 228N in patients with thrombocytosis strongly suggests CMPD. On the other hand, the absence of hyperploid cells > 128N or the presence of a small fraction (less than 5%) of moderately hyperploid cells (96-228N) indicates reactive thrombocytosis. Image cytometry analysis therefore appears to add valuable information to conventional cytological examination in the diagnostic assesment of thrombocytosis. Conclusions. S-Form software image analysis is much faster and more reliable than microdensitometric DNA analysis, which we previously used for cytometric studies. We believe the method of image DNA analysis has the potential for widespread clinical use in the differential diagnosis of megakaryocyte disorders.
Summary		Izhodišča. Število, velikost in ploidnost megakariocitov so odločilni kazalci za diagnozo mieloproliferativnih bolezni (MPB). Pri reaktivnih stanjih najdemo občasno v kostnem mozgu podobne spremembe, ki jih z dostopnimi metodami težko razlikujemo od tistih pri MPB. Z novo metodo slikovne citometrije smo določali razliko v ploidnosti megakariocitov pri nekaterih MPB in reaktivnih trombocitozah. Metode. Z metodo slikovne citofotometrije in novim računalniškim programom S-Form smo analizirali aspirate kostnega mozga 11 bolnikov z reaktivno trombocitozo in 12 bolnikov s klonalno trombocitozo pri MPB. Za barvanje smo uporabili modificirano Feulgenovo metodo. Pri vsakem bolniku smo analizirali po 100 megakariocitov in izračunali delež hiperploidnih celic čez 96N in 128N. Rezultati. Prisotnost > 5% megakariocitov s ploidnostjo več kot 96N ali >= 2% megakariocitov s ploidnostjo več kot 128N pri bolniku s trombocitozo govori v prid MPB. Nasprotno odsotnost hiperploidnih celic > 128N ali prisotnost malega deleža (manj kot 5%) celic s ploidnostjo med 96N in 128N kaže na reaktivno trombocitozo. Zaključki. Slikovna analiza DNK megakariocitov z računalniškim programom S-Form je koristna in praktična metoda za razlikovanje primarnih in reaktivnih trombocitoz. Menimo, da bo pomemben dodatek pri citološki diagnostiki posameznih megakariocitnih motenj.
Descriptors		THROMBOCYTOSIS IMAGE CYTOMETRY MEGAKARYOCYTES PLOIDIES DIAGNOSIS, DIFFERENTIAL