OBJECTIVE

The aim of this study was to evaluate the analytical performance of a new portable haemoglobinometer, Hemo_Control (EKF-diagnostic, GmbH, Germany), which measures haemoglobin concentration in venous and capillary samples.

METHODS

The within series and between series imprecision of the Hemo_Control instrument were calculated after measuring the concentration of venous samples under standardized conditions; by experienced laboratory technicians in a hospital laboratory, and venous and capillary samples under conditions similar to where the instrument is intended for use; by personnel at two primary health care centres. The bias of the Hemo_Control instrument was calculated as the difference between its results and results obtained with a Coulter LH 750 instrument traceable to the ICSH reference method.

RESULTS

The uncertainty of the Hemo_Control instrument for venous samples was lower than the quality goal of +/- 5% considered acceptable for patient care. High within series imprecision (5.5%) was observed for measurements of capillary blood samples in one of the primary care centres, whereas adequate analytical performance was obtained at the other centre. The Hemo_Control instrument showed negligible bias of +0.8 g/L for both venous and capillary samples in primary health care.

CONCLUSIONS

The observed uncertainty indicates that Hemo_Control is appropriate for near patient testing using venous samples. Capillary samples may be used if sampling skills are adequate.

Standardization and harmonization in Laboratory testing are a key issue in the midst of globalization era, because most of laboratory testing has been currently achieved with various kinds of automated systems. In the developed countries, automated systems with highly-regulated principles are commonly used in the routine laboratory. However, there are so many undeveloped and developing countries in Asia that diversity of testing levels can be observed in the area. Some laboratories use glass chamber method for blood cell counting, while other laboratory use semi-automated or fully automated analyzers for complete blood count. International standardization on Hematology is focused on the developed system but not for the developing system. Established standardized documents therefore whould not be unsuitable for Asian societies. In the context, International Council for Standardization in Hematology (ICSH) changed its rules to adjust our Asian Societies and ICSH started to restructure the body. International ICSH society is divided into 5 region sub-groups. Asian area is able to possess one new sub-society, ICSH-Asia. Its reconstruction work has been just started with Asain colleagues, and we are now extending the new societies to discuss Asian problems on the quality of hematology testing.

BACKGROUND

The CellaVision Advanced Red Blood Cell (RBC) Software Application is a new software for advanced morphological analysis of RBC, which automatically performs a preliminary characterization and grouping of RBC into 21 morphological categories, including schistocytes. Upon automated classification, the software offers the possibility of reclassification of RBC by the operator. The aim of this study was to evaluate the schistocyte analysis by the CellaVision Advanced RBC Application.

METHODS

Schistocyte counts were evaluated comparing the automated count on a CellaVision DM96, both before and after reclassification, with the reference manual microscopic method according to the ICSH criteria. Thirty-six samples of hospitalized patients and 40 samples of controls were analyzed.

RESULTS

Within-run, between-run and between-observer coefficients of variation were lower when counted with the CellaVision compared to the manual microscopic count. The very high sensitivity but rather poor specificity implicates the need for reclassification by the operator, following automated analysis. After reclassification, method comparison studies revealed good agreement with the manual microscopic method, with however slightly higher values of schistocytes for the automated analysis.

CONCLUSIONS

The CellaVision Advanced RBC Software Application provides a sensitive and reproducible measurement of schistocytes in peripheral blood, but still requires manual revision. Furthermore, it is an easy-to-use software and an excellent teaching tool that might contribute to standardization in the investigation of schistocyte-related conditions.

OBJECTIVE

Diagnosis of a case with congenital dyserythropoietic anemia (CDA).

METHODS

Routine tests for hemolysis were carried out. The activities of erythrocyte enzymes were measured according to the methods recommended by international committee for standardization in hematology (ICSH). The quantity and quality of erythrocyte membrane proteins were analyzed with 4% - 15% sodium dodecyl sulfate-polyacrylamide gradient gel electrophoresis (SDS-PAGE). The membrane ultrastructure of erythrocyte from bone marrow was observed under transmission electron microscope (TEM).

RESULTS

The main results were: (1) Bone marrow morphology revealed erythroid hyperplasia and 0.10 symmetrically binucleated late erythroblasts. The erythrocytes in peripheral blood showed anisocytosis and hypochromia. (2) The intracellular iron was 0.98 and the storage iron was strongly positive in bone marrow. The serum ferritin was 1607 micro g/L. The content of blood sugar was 27.5 mmol/L. (3) Ham test was negative in his own acidified serum but positive in the group-compatible sera. (4) A quick mobile H band was seen in hemoglobin electrophoresis. H inclusion test was positive. (5) SDS-PAGE demonstrated that the migration of band 3 protein of erythrocyte membrane in an electric field was faster (110%) than that of normal controls and the relative contents of band 1, band 3, band 4.1 were reduced to various extent. (6) "Double membrane" with gap and shedding was observes under TEM.

CONCLUSIONS

The final diagnosis of the case was CDAII, also called HEMPAS (hereditary erythroblastic multinuclearity with positive acidified serum), accompanied with alpha thalassemia and secondary siderosis and diabetes.

The schistocytes are fragmented red blood cells mainly observed in the setting of hemolytic anemias and particularly among the thrombotic microangiopathies. The presence of schistocytes is an important criterion for the diagnosis of mechanical anemias, though the identification of these cells remains problematic. As a high variability of the morphologic identification criteria of the schistocytes among morphologists has been observed, some guidelines have been proposed after workshops (French and Italian groups). The International council for standardization in hematology published a consensus in November, 2011. The French group of cellular hematology (GFHC) aimed to recover the opinion of French biologists directly confronted to schistocytes measurements. 169 out 500 (34%) answered 10 questions dealing with the identification and measurements of schistocytes as proposed by the ICSH guidelines. A consensus was reached for the urgent need of guidelines documents, moreover in the current background of the European accreditation NF EN ISO 15189 rules. A traduction in native (French) language as warmly wished in order to facilitate the diffusion of the information. New fragmented red cell parameter recently provided by 2 manufacturers of automated blood cell counters remained doubtfull for routine use for half of the biologists.

In assigning values to fresh blood specimens used for calibrating automated blood cell analysers, haemoglobin concentration should be determined by the ICSH recommended hemiglobincyanide (HiCN) method. Errors in the reference method may be caused by inadequate blood collection, e.g. incorrect anticoagulation; inadequate dilution, e.g. use of inadequately calibrated pipettes; incomplete conversion of haemoglobins to HiCN, e.g. insufficient time for reactions to come to completion; during measurement, e.g. due to turbidity of the HiCN solution to be measured; or by transcription error. Readily available commercial reagent systems and secondary standards are compared with ICSH standards as to their compliance with these standards.

The effects of partial adenohypophysectomy on comb size, testicular weight, and histology 20 or 30 days after the operation are described and discussed in relating these observed changes with the volume of remaining adenohypophyseal tissue. From the present observation, it is concluded that 1) the response of the domestic fowl to partial deficiency of adenohypophyseal tissue may be more sensitive than that of some mammals, 2) the decrease in testicular weight and tubular diameter precedes the histological change in testis and comb regression, and 3) to maintain fully active testicular function, more than 75% of the adenohypophysis is needed. About 40% of the adenohypophyseal tissue including a remnant of cephalic lobe can stimulate spermatogenesis, but the spermatozoa produced were partially abnormal. The critical amount of adenohypophyseal tissue for normal spermiogenesis may exist between 55 and 75%. All histological change is initially observed in the center of testis and then extended toward the periphery. It is suggested that the cephalic lobe contains at least a trace of ICSH in addition to FSH.

The Cell-Dyn 1000 is a new eight parameter fully automatic cell counter. This was evaluated following the ICSH guidelines. Scientific evaluation was generally satisfactory, largely conforming to the manufacturer's specifications. A number of potential microbiological safety hazards were noted; many could be overcome by minor modifications. Throughput and start-up/close-down times were acceptable. However, the reliability of the tested machine was felt to be less than satisfactory although this may be corrected by further minor machine modifications.