A viable alternative to the ICSH reference method for serum iron assay is proposed where automated batch analysis is required. The introduction of Ferrozene into contemporary automated methodology makes this possible. The Donnan effect on membrane dialysis has been resolved by observing the fate of aqueous and protein matrices inoculated with 59FeCl3 as they leave the system.

In order to determine if mutant hemoglobins can be identified by relatively simple methods, a Working Group of the ICSH Expert Panel on Abnormal Hemoglobins and Thalassemia analyzed 17 hemolysates containing 14 different mutant hemoglobins by four electrophoretic methods: (1) cellulose acetate in alkaline buffers, (2) citrate agar pH 6.0, (3) urea 2-mercaptoethanol buffer pH 8.9, and (4) urea 2-mercaptoethanol buffer pH 6.0. The examined mutants included several of great numerical and clinical importance as well as some rare ones, namely, HbS, C, D Los Angeles (Punjab), E, G Philadelphia, N Baltimore, and O Arab; also Hb Ft. Worth, Montgomery, Winnepeg, Rush, Q India, Bethesda, and Lepore. Comparative mobilities of these hemoglobins in all of the methods are presented here. The combined data permit their presumptive identification, often with a high degree of specificity. The system has been applied in Iran, where the four prevalent mutants can be differentiated by these methods, at considerable saving of time and resources previously expended on structural analyses. It is proposed as a basis for an ICSH Tentative Standard. There is little doubt that this presumptive identification of hemoglobin variants by simple electrophoresis will be improved or complemented by the introduction of newer techniques, such as immunologic analysis. However, for the present and for some time to come, the system outlined here should be found valuable. The present report does not concern itself with the numerous auxiliary techniques involved in the identification of abnormal hemoglobins-sickle-cell test, solubility tests, lability test--and no claim is made that the simple system described here eliminates these other techniques from the diagnostic armamentarium of the laboratory.

Before a new method is used for clinical testing, it is essential that it is evaluated for suitability for its intended purpose. This document gives guidance for the performance of verification, validation and implementation processes required by regulatory and accreditation bodies. It covers the planning and execution of an evaluation of the commonly performed screening tests (prothrombin time, activated partial thromboplastin time, thrombin time and fibrinogen assay), and instrument-specific issues. Advice on selecting an appropriate haemostasis analyser, planning the evaluation, and assessing the reference, interval, precision, accuracy, and comparability of a haemostasis test system are also given. A second companion document will cover specialist haemostasis testing.

Keywords: haemostasis; laboratory automation; precision.

Rapid and accurate analysis of platelet count plays an important role in evaluating hemorrhagic status. Therefore, we evaluated platelet counting performance of a hematology analyzer, Celltac F (MEK-8222, Nihon Kohden Corporation, Tokyo, Japan), that features easy use with low reagent consumption and high throughput while occupying minimal space in the clinical laboratory. All blood samples were anticoagulated with dipotassium ethylenediaminetetraacetic acid (EDTA-2K). The samples were stored at room temperature (18(;)C-22(;)C) and tested within 4 hours of phlebotomy. We evaluated the counting ability of the Celltac F hematology analyzer by comparing it with the platelet counts obtained by the flow cytometry method that ISLH and ICSH recommended, and also the manual visual method by Unopette (Becton Dickinson Vacutainer Systems). The ICSH/ISLH reference method is based on the fact that platelets can be stained with monoclonal antibodies to CD41 and/or CD61. The dilution ratio was optimized after the precision, coincidence events, and debris counts were confirmed by the reference method. Good correlation of platelet count between the Celltac F and the ICSH/ISLH reference method (r = 0.99, and the manual visual method (r= 0.93) were obtained. The regressions were y = 0.90 x+9.0 and y=1.11x+8.4, respectively. We conclude that the Celltac F hematology analyzer for platelet counting was well suited to the ICSH/ISLH reference method for rapidness and reliability.

Warfarin has been widely used for an oral anticoagulant therapy against thrombotic diseases. For the monitoring of its anticoagulant intensities, prothrombin time (PT) ratio and percentage of thrombotest (TT) are commonly used in Japan. Recently, International Normalized Ratio (INR) was recommended by ICSH/ICTH. Practicality and usefulness of INR and its combined use of thrombin-antithrombin III complex (TAT) for the monitoring of oral anticoagulation therapy were evaluated among patients of ischemic heart disease with or without interventions, and of cardiomyopathies and valvular diseases. Difference in thromboplastin sensitivities have been shown to cause errors in PT elongation and in the evaluation of anticoagulant activity, so that the monitoring only by PT ratio is considered to be irrelevant, and that INR is recommended to be used. INR was comparable to the levels of TT. Majority of the patients, whose TAT levels were kept normal, were controlled below the proposed therapeutic ranges of INR. With the combination of INR and TAT monitoring, anticoagulant effect of warfarin could be achieved safer in lower dose than the levels that might cause bleeding accidents.

Since rapid blood count analysis as near patient testing is expanding, we evaluated the use of a Sysmex pocH-100i compact haematology analyser in an outdoor oncology setting according to the recently published International Council for Standardization in Haematology (ICSH) guidelines. In total, 838 blood samples from oncology patients were analysed by pocH-100i and re-analysed by a high-throughput haematology analyser for comparison (Abbott CD-4000 or Sysmex XE-2100) to evaluate in use imprecision, comparability and vote-outs. Imprecision was less than 5%, except for platelet enumeration in the low range (within-run imprecision 7%). Good comparability was found even for platelet enumeration in the low range (r2 = 0.82). Vote-outs were found in 10.6% of examined samples. In conclusion, the Sysmex pocH-100i demonstrates good imprecision conform with former publications, produces reliable results in normal and in lower ranges comparable to the results of high throughput haematology analysers. In a well controlled management plan the Sysmex pocH-100i is suitable for near patient testing in oncology.

OBJECTIVE

To show distribution of the investigated patients into diagnostic groups, find out the diagnostic value of the levels of hemoglobin and packed cell volume as possible markers of absolute erythrocytosis in the group of patients with polycythaemia.

METHODS

We evaluated 61 patients, mean age was 46 years (18-82), 9 females and 52 males before treatment. Mean levels of hemoglobin in females - 171 g\l (143-190), packed cell volume 52% (49-61). Mean levels of hemoglobin and packed cell volume for males were 187 g/l (168-196) and 57,8% (49-65), respectively. All blood samples were taken in the morning. Full blood picture of venous blood was determined by Coulter principle on Gen S ("Beckman-Coulter", USA) blood analyzer with preserving agent (ethylene diamine tetraacetate, EDTA). Red cell mass and plasma volume were measured by the radionuclide method (Cr-51). Results were performed with an allowance for patient's surface area and were interpreted according to International Council for Standardization in Haematology guidelines (ICSH) (Pearson et al. 1995).

RESULTS

Polycythaemia vera was detected only in 19 (31%) among 61 patients, 15 patients refused from further investigation. Among others 46 patients 14 subjects had secondary erythrocytosis, among them 9 were diagnosed with absolute erythrocytosis (hypoxic) and 5 with idiopathic erythrocytosis. Relative ("apparent") erythrocytosis was detected in 13 cases. Measurement of red cell mass allowed us to divide patients into groups with absolute and relative erythrocytosis. Such laboratory parameters as hemoglobin, number of red blood cells and packed cell volume do not always completely show the level of red cell mass due to possible variations of the plasma volume and can not be the reason for diagnosis of haematological disorder. It is shown that hemoglobin level over 185 g/l confirms the presence of absolute erythrocytosis only in 50% of males with polycythaemia, 15% of males with secondary erythrocytosis might have incorrect diagnosis as though increased red cell mass. Statistically defined highly significant (p=0,001) difference of the level of red cell mass in males with polycythaemia and patients with "apparent" polycythaemia turned out to 166% and 111%, respectively. The levels of red cell mass in patients with polycythaemia confirm absolute erythrocytosis over superior normal limit (more then 25%) in comparison with secondary erythrocytosis where red cell mass rate remained normal. Average plasma volume measurements in the same groups of patients were at normal range - 95% u 81%, respectively. Difference between these mean values was authentically significant.

CONCLUSIONS

Red cell mass and plasma volume measurement is easy and necessary procedure for estimation absolute and "apparent" polycythaemia. Rather common occurrence of different forms of erythrocytosis and in particular "apparent" erythrocytosis must determine certain diagnostic approach according to specific clinical case.

The partial thromboplastin time (PTT) test is widely used as a screening test for the detection of hemophilia. It is also used to monitor patients on heparin anticoagulation. This proposal from a ICSH Panel proposes guidelines for the performance of this test, including comparable reference ranges, precision and sensitivity requirements.

A simple accurate and very reproducible procedure for measuring serum iron concentration without precipitating serum proteins is described. In the same time, iron is released from serum transferrin, reduced and determined at 535 mn by incubation in a water bath at 55 degrees for five minutes within a formate buffered system (pH 2,6; i = 0,05) including bathophenanthroline sulfonate. This method yields values very similar to those obtained by that of the International Committee for Standardization in Hematology (ICSH).

BACKGROUND

Measurement of the length of sedimentation reaction in blood (LSRB), also called erythrocyte sedimentation rate (ESR), is a widely used hematology test. This study intends to compare ESR levels measured by Test-1 method and International Council for Standardization in Hematology's (ICSH) reference method, and analyzes the effect of hematocrit (Hct) on ESR results.

METHODS

A total of 755 patients from 2 hospitals were included in the study, and samples with EDTA were studied by Test-1 method for ESR measurement and total blood count, whereas citrated samples were studied with reference Westergren method. Then, 2 methods were compared. Distribution of ESR results according to the ESR(≤20, >20 mm/h) and Hct(≥35%, <35%) levels and hospital type was analyzed. ESR levels with Hct levels<35% were corrected with Fabry's formula.

RESULTS

The mean and SD values for the Test-1 method, reference Westergren method, and corrected ESR measurement were 21.30 ± 18.39, 28.59 ± 25.82, and 24.92 ± 20.58 mm/h, respectively. Within the whole group, the correlation coefficient (r) was .77 (.7-.80) with a significance level P < .001. Passing-Bablok regression analysis of the methods resulted in a regression equation y = 1.00 (95% Cl: 0.43-1.88) + 0.75 (95% Cl: 0.70-0.78)x while the significance of linearity was acceptable (P < .01). All subgroup linear regression analyses revealed that the correlation was acceptable, except ESR>> 20 mm/h group, Hct < 35% group, and corrected ESR group (significance level were P>> .10).

CONCLUSIONS

The study showed that the role of the hospital and the capacity of testing are important in choosing the instrument for measuring ESR. Furthermore, the patient profile, especially malignancy possibility and Hct level, may be important for instrument selection.

Background: The Advanced RBC Application of the CellaVision DM9600 system (CellaVision AB, Lund, Sweden) automatically characterizes and classifies red blood cells (RBCs) into 21 morphological categories based on their size, color, shape, and inclusions. We evaluated the diagnostic performance of the CellaVision Advanced RBC Application with respect to the classification and grading of RBC morphological abnormalities in accordance with the 2015 International Council for Standardization in Haematology (ICSH) guidelines.

Methods: A total of 223 samples, including 123 with RBC morphological abnormalities and 100 from healthy controls, were included. Seven RBC morphological abnormalities and their grading obtained with CellaVision DM9600 pre- and post-classification were compared with the results obtained using manual microscopic examination. The grading cut-off percentages were determined in accordance with the 2015 ICSH guidelines. The sensitivity and specificity of the CellaVision DM9600 system were evaluated using the manual microscopic examination results as a true positive.

Results: In pre-classification, >90% sensitivity was observed for target cells, tear drop cells, and schistocytes, while >90% specificity was observed for acanthocytes, spherocytes, target cells, and tear drop cells. In post-classification, the detection sensitivity and specificity of most RBC morphological abnormalities increased, except for schistocytes (sensitivity) and acanthocytes (specificity). The grade agreement rates ranged from 35.9% (echinocytes) to 89.7% (spherocytes) in pre-classification and from 46.2% (echinocytes) to 90.1% (spherocytes) in post-classification. The agreement rate of samples with within-one grade difference exceeded 90% in most categories, except for schistocytes and echinocytes.

Conclusions: The Advanced RBC Application of CellaVision DM9600 is a valuable screening tool for detecting RBC morphological abnormalities.

Keywords: Advanced RBC Application; CellaVision DM9600; International Council for Standardization in Haematology guidelines; Morphology; Red blood cell.