ALBERT

Description: Introduction: The diagnostic criteria for polycythemia vera (PV) has recently been updated by the World Health Organization (WHO). The criterion for erythrocytosis has been modified downwards: hemoglobin (hb)〉 16.5 g/dL or hematocrit (hto)〉 49% in men and hb〉 16 g/dL or hto〉 48% in women. This reduction increases the potential number of patients that would be test for JAK2 V617F mutation if PV is suspected. The V617F mutation in the JAK2 gene is present in 95% of cases of PV. It is estimated that the prevalence of this mutation in the general population is around 0.2%. Our aims are to determine the prevalence of JAK2 V617F in individuals with erythrocytosis according to WHO2016 criteria and to find prognostic factors that could help to identify patients with PV. Methods: We prospectively studied all hemograms performed in our laboratory during 7 nonconsecutive days. Variables studied were hb, hto, leukocytes, neutrophils, platelets, MCV, MCH, MCHC and RDW. JAK2 V617F mutation was studied in all males that had hb〉 16.5 g/dl or hto〉 49% or females that had hb〉 16 g/dl or hto〉 48%. JAK2 V617F mutation was studied by PCR assay in which an amplification control fragment and the JAK2 mutant allele were simultaneously amplified. All positive samples were confirmed by quantitative real-time PCR in a reference laboratory. Positive results were considered when the JAK2 V617F allele ratio was ≥ 0.7. The variables collected were correlated with the result of the JAK2 test in a univariate way. The T-Student test was used for the quantitative variables and the Chi-square test for the categorical variables. For the cell count variables, the Mann-Whitney U test was used. Results: A total of 15366 HG were analyzed. 1271 (8.3%) met the inclusion criteria for erythrocytosis. JAK2 V617F was performed on 1001 samples (270 samples were not suitable for the PCR assay due to low quality). Twelve samples (1.2%) were positive for JAK2 V617F mutation. However, 5 samples were excluded due to a known diagnosis of myeloproliferative neoplasm. Therefore, finally prevalence of JAK2 V617 mutation in 996 patients that met WHO erythrocytosis criteria was 0.8% (8/996). Medians for all parameter studied for each group are shown in table 1. In order to find out parameters that could increase the incidence probabilities to identify patients with JAK2 V617F we performed an univariate analysis of the variables included, according to JAK2 mutational status. We found that patients with JAK2 V617F had higher levels of leukocytes, neutrophils, platelets and RDW than patients with negative JAK2 (p

Description: INTRODUCTION: The 2016 update of the World Health Organization (WHO) classification criteria for Polycythemia Vera (PV) included a decrease of the hemoglobin threshold. This modification increases exponentially the individuals with potential PV. International groups have proposed diagnostic algorithms for the screening of individuals with potential PV. However, these algorithms are not based on the prevalence of JAK2V617F in this population, and to our concern has not been prospectively validated. Our aim is to develop and validate an algorithm based on previously reported prevalence of JAK2V617F in individuals with potential PV in our environment. METHODS: This study was designed in two phases: a first phase for the development of the algorithm and a second phase for the validation of the algorithm. Phase 1: from previous data collected from our group of JAK2 V617F prevalence in individuals with erythrocytosis according to WHO criteria we performed an area under the curve (ROC) analysis with optimal cutoff point in order to maximize sensibility and specificity of the collected variables. The thresholds for platelets and neutrophils were selected for being clinically relevant and with an AUC 〉0.8. With these two variables and their optimal cutoff point, a sequential 2-step-algorithm was designed (figure 1). Phase 2: for the validation of the algorithm, hemograms from outpatient individuals were selected, and the "2-step-algorithm" was applied. Therefore, if samples met the criteria for step 1 (hemoglobin 〉 16.5 g/dl for men or 16 g/dl for women or hematocrit 〉49 % for men or 48 % for women) then, the step 2 was applied (platelets 〉 248x103/ml or neutrophils〉5.98x103/ml). Finally, all samples that passed the 2-step-algorithm, were tested for JAK2 V617F. With the chosen samples a JAK2 V617F qualitative PCR with a sensibility of at least 〉0.1% was performed. The positive results were validated and quantified in our reference laboratory, and those with an allele burden 〉1% were considered positive. RESULTS: A total of 15298 hemograms were initially selected. Of those, 1595 (10.4%) met the erythrocytosis criterion (step 1). 581 (36%) of these met the step 2 criteria and the JAK2 V617F PCR was performed in 501 samples. A total of 7 positive cases (1.4%) were found, none of which was previously diagnosed of myeloproliferative neoplasm. The median of hemoglobin and hematocrit was 16.3 g/dl and 50.3% respectively. Of the selected samples, 2 of them met the platelet criterion, 4 of them the neutrophils criterion and 1 of them met both. After reviewing clinical records, 43.8% of them were found to have cardiovascular events history history: 1 patient had atherosclerosis of the lower limbs, 1 had acute coronary syndrome and 1 had an ischemic stroke. Characteristics of JAK2V617F individuals are detailed in table 1. Comparing to step 1, the addition of step 2 increases the efficiency of the algorithm 1.75 times. In order to know the prevalence of JAK2 V617F clonal hematopoiesis in our environment, 300 unselected samples were tested. We found 1 positive result. However, after reviewing clinical records, this sample was found to belong to a patient with known myeloproliferative neoplasm. Thus, JAK2 V617F prevalence in our environment was considered