Clinical Value of Hypochromia Markers in the Detection of Latent Iron Deficiency in Nonanemic Premenopausal Women.

BACKGROUND: Iron deficiency (ID) is the most common cause of anemia in fertile women and hemoglobin (Hb) within the reference interval does not exclude ID. The consequence of an imbalance between the iron requirements and supply is a reduction of red-cell Hb content, which causes hypochromic cells. The aim of this study was to assess the reliability of new parameters low Hb density (LHD%), reticulocyte Hb equivalent (RetHe), and percentage of hypochromic erythrocytes (%HypoHe) in the detection of latent ID (LID), defined as depletion of iron stores without anemia. METHODS: Two hundred fifty consecutive nonanemic women in fertile age (18-40 years, mean 33.5 years), whose analyses had been requested by general practitioners, were included. Independent samples t-test, receiver operating characteristic (ROC) curve analysis (gold standard for detecting LID ferritin <30 µg/l), and Cohen's kappa index were applied. RESULTS: One hundred fifty-three women had ferritin within the reference range and Hb >120 g/L; 97 (38.8%) had LID. The results were as follows: %HypoHe-AUC 0.934, cutoff 1.6%, sensitivity 85.7%, specificity 92.1%; RetHe-AUC 0.914, cutoff 29.9 pg, sensitivity 86.8%, specificity 85.7%; LHD%-AUC 0.898, cutoff 5.0%, sensitivity 85.9%, specificity 84.1%. Applying those cutoffs, agreement between ferritin and %HypoHe was κ 0.61 and 0.56 for RetHe and LHD%. CONCLUSIONS: LHD%, %HypoHe, and RetHe emerge as reliable tests for the investigation of LID and could improve the ability to detect ID before anemia is present.

Biomarkers of hypochromia: the contemporary assessment of iron status and erythropoiesis.

Iron status is the result of the balance between the rate of erythropoiesis and the amount of the iron stores. Direct consequence of an imbalance between the erythroid marrow iron requirements and the actual supply is a reduction of red cell hemoglobin content, which causes hypochromic mature red cells and reticulocytes. The diagnosis of iron deficiency is particularly challenging in patients with acute or chronic inflammatory conditions because most of the biochemical markers for iron metabolism (serum ferritin and transferrin ) are affected by acute phase reaction. For these reasons, interest has been generated in the use of erythrocyte and reticulocyte parameters, available on the modern hematology analyzers. Reported during blood analysis routinely performed on the instrument, these parameters can assist in early detection of clinical conditions (iron deficiency, absolute, or functional; ineffective erythropoiesis, including iron restricted or thalassemia), without additional cost. Technological progress has meant that in recent years modern analyzers report new parameters that provide further information from the traditional count. Nevertheless these new parameters are exclusive of each manufacturer, and they are patented. This is an update of these new laboratory test biomarkers of hypochromia reported by different manufactures, their meaning, and clinical utility on daily practice.

Erythrocyte and reticulocyte parameters in iron deficiency and thalassemia.

INTRODUCTION: Red blood cells (RBCs) extended parameters or erythrocyte subsets are now reported by the new Sysmex XE 5000 analyzer. This study was aimed at establishing a characteristic analytical feature, including the new erythrocyte and reticulocyte parameters, in case of thalassemia trait and iron deficiency (IDA). METHODS: Ninety healthy individuals, 136 ß-thalassemia carriers, 121 mild IDA, and 126 severe IDA patients were analyzed. RESULTS: The values obtained for the RBC extended parameters were significantly different (P<0.0001) in the groups; the only exception was %Hypo-He in the case of mild IDA and thalassemia (P=0.6226). %Hypo-He was considerably greater in severe IDA (23.4%) than in mild cases (12.4%), P<0.0001. %MicroR was more increased in thalassemia (38.6 %) than in the mild IDA (16.5%, P<0.001) and in severe IDA (21.6%, P<0.001). Immature reticulocyte fraction (IRF) mean values in the groups were statistically different; the thalassemia group had an intermediate value (8.7%) between healthy (4.4%) and IDA (16.7 and 12.9%). CONCLUSIONS: Erythrocytosis and severe microcytosis, together with a high percentage of microcytes and a moderate increase in IRF, is the profile of ß-thalassemia carriers, whereas anisocytosis and the hypochromic subset correlates with the severity of the anemia in iron-deficient patients.

The role of automated measurement of RBC subpopulations in differential diagnosis of microcytic anemia and ß-thalassemia screening.

Cell counter-based formulas have been used in the differential diagnosis of microcytic anemia. The measurement of RBC subpopulations is now available on the Sysmex XE 5000 analyzer (Sysmex, Kobe, Japan). We describe the new formulas: % microcytic - % hypochromic; and % microcytic - % hypochromic - red cell distribution width (RDW), derived from the percentages of microcytic and hypochromic RBCs. The present study aimed to prospectively evaluate the reliability of these new formulas in the differential diagnosis of microcytosis and ß-thalassemia screening compared with already published indices. The indices were calculated for a set of 250 iron-deficient patients and 270 ß-thalassemia carriers. Independent samples t test and receiver-operating characteristics analysis were applied. The % microcytic - % hypochromic - RDW, % microcytic - % hypochromic, and Green and King indices provided higher areas under the curve. The % microcytic - % hypochromic - RDW was the most reliable index evaluated, with 100% sensitivity and 92.6% specificity. This index can be used to efficiently screen patients with microcytosis for further hematologic studies to confirm ß-thalassemia.

Percentage of hypochromic erythrocytes as a potential marker of iron availability.

BACKGROUND: The percentage of hypochromic red cells (% Hypo-He) is now available on the Sysmex analyzer XE 5000 (Sysmex Corporation, Kobe, Japan). In this study the reliability of % Hypo-He in the assessment of iron status in the presence of inflammation is investigated. METHODS: Ninety healthy subjects, 91 patients with iron deficiency anemia (IDA) previous to therapy, 65 patients with chronic kidney disease (CKD), 57 patients with anemia of chronic diseases (ACD) and 28 patients with storage iron deficiency (ACD/IDA) receiving therapy were analyzed. Pearson's correlation, Independent samples t-test, receiver operating characteristic (ROC) curve analysis and Cohen's k index were applied. RESULTS: % Hypo-He values inversely correlated with the blood hemoglobin (Hb) (r=-0.78) and Hb content (MHC, r=-0.75). In patients with IDA and ACD/IDA the values obtained for % Hypo-He showed no statistical difference (p=0.5037). ROC analysis results for % Hypo-He in the detection of iron deficiency defined by a serum transferrin receptor (sTfR) >21 nmol/L. The area under the curve is 0.929; cut-off 3.6%, sensitivity 91.5% and specificity 80.9%. Using the threshold 3.6% the k index obtained in comparison to sTfR was 0.64. CONCLUSIONS: % Hypo-He is a reliable parameter for the detection of iron deficiency in patients with anemia in the presence of inflammation.

Erythrocyte and Reticulocyte Indices on the LH 750 as Potential Markers of Functional Iron Deficiency.

Reticulocyte hemoglobin content (CHr) and percentage of hypochromic cells (%Hypo) are restricted to the Siemens analysers. The aims of the study were to investigate the correlation of Red cells size factor (RSf) and low Hemoglobin density (LHD%), reported by Beckman-Coulter analysers, with CHr and %Hypo in the assessment of iron status in the presence of inflammation. 381 samples were run on both LH 750 (Beckman-Coulter) and Advia 2120 (Siemens) analysers. Correlation between parameters were calculated and the diagnostic performance of the new parameters was assessed. Results. Correlation between RSf and CHr, r = 0.85. ROC curve analysis for RSf in the diagnosis of iron restricted erythropoiesis defined as CHr < 28 pg: AUC 0.983; Cutoff 91.1%; Sensitivity 98.8%; Specificity 89.6% Correlation between LHD% and %Hypo, r = 0.869. ROC curve analysis for LHD% in the diagnosis of iron deficiency defined by %Hypo >5%: AUC 0.954; Cut off 6.0%; Sensitivity 96.6%; Specificity 83.2% Conclusions. RSf and LHD% could be reliable parameters for the study of iron metabolism status.

Potential utility of the new Sysmex XE 5000 red blood cell extended parameters in the study of disorders of iron metabolism.

BACKGROUND: New erythrocyte parameters are reported by the Sysmex XE 5000 analyzer. This instrument measures the hemoglobin (Hb) content of individual red cells, calculates the percentage of hypochromic red cells (%Hypo He) and the percentage of hyperchromic red cells (%Hyper He) and quantifies the proportion of marginally sized erythrocytes (%Micro R and %Macro R). The goals of the study were to establish the reference range for erythrocyte extended parameters, their value in different types of anemia and to investigate their reliability in the study of disorders of iron metabolism. MATERIALS: Three hundred and ninety samples were analyzed. The Kolmogorov-Smirnoff test, independent samples t-test and Pearson correlation were calculated; receiver operating characteristic (ROC) curve analysis was used to determine their diagnostic performance. RESULTS: The values of the four parameters studied were normally distributed and statistically different (p<0.0001) in the different groups of patients; the only exception was %Hypo He in cases of iron deficiency and thalassemia (p=0.3758). Results of ROC curve analysis for %Hypo He in the diagnosis of restricted erythropoiesis reticulocyte Hb content (reticulocyte hemoglobin content, Ret He <29 pg) were: area under the curve 0.963; cut-off 1.8%; sensitivity 98.3%; specificity 91.1%. CONCLUSIONS: The new parameters appear to be sensitive for detecting small changes in the number of red cells with inadequate hemoglobinization and volume.