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Analitical validation and clinical significance of determinati on (rPl%) reticulated platelet on hematology analyzer-Abbott-Sapphire

Introduction: Small proportion of circulating platelets, which are characterized by residual, probably the reticular cytoplasmic RNA are reticulated platelets (rPl). Simultaneously, the development of technology has allowed the determination of the rPl % on hematology analyzers, but for varying reference interval depending on the technology, clinical application remains questionable. The aim of study was : 1.to investigate intra-assay CV and reference range limits in physiological conditions for the determination of rPl% on Abbott CD Sapphire analzyer, 2.to investigate if there were difference in the value of rPl % between healthy subjects and a.patientes with post-chemotherapy thrombocitopenia -Group I ; newborns with ITP- Group II ; patients with depressive disorder and low value of platelet serotonin-Group II. Methods: Peripheral blood samples were analysed on Abbott CD Sapphire. The study included 174 healthy and sex –matched individuals (median age 50, 06 ± 11, 84 SD).Group I, include 50 patients with hematologic neoplasms after chemotherapy treatment and platelet counts less then 50 x 10 9/L ; Group II, include 12 newborns with ITP and positive anti- platelet antibody tests ; Group III, include 137 patients with depressive disorder and statistically significantly lower platelet serotonin (259±175.92ng/10 9platelets). Results: The duplicate analysis of forty samples from healthy individuals have acceptable value of reproducibility ; mean CV = 17, 3% ; for both subgroups according to their mean rPl % (mean rPl=1, 66 ; CV = 18% ; mean rPl =2, 76 ; V =16, 6).The median platelet count in 174 healthy controls was 238, 7 x10 9 /L and rPl % values showed non-parametric disribution (Kolmogorov - Smirnov test ). The reference interval of healty individuals presented as range of 2, 5-97, 5 percentile with 95% CI, was 0, 45-4, 56 %. We found a stastically significant difference (p< 0.001) in comparation of healthy subjects and heterogeneous group of thrombocitopenic hematology patientes (median 11, 1 %) and newborns with ITP (median 14, 2 %).We found statisticly siggnificant difference for propotion of reticulted pletelets in patients with depressive disorder (2, 36±1, 38 %) in comparasion to healthy (1, 58±0, 722 %). Conclusions: The validation of method for determination of rPl on Abbott Sapphire hematology analyzer was carried out and reference range limits shown in pysiological condition: lower limit : o, 45%, upper limit: 4, 56% of reticulated platelets- rPl%.We found a statistically significant excess of rPl in hematologic patients with thrombocytopenia, which probably refers to the remission. The patients with depressive disoder and reduced platelet serotonin concentration showed an increased prevalence of rPl compared to the healthy population.

reticulated platelets; analitical validation; residual cytoplasmic RNA

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