BACKGROUND

An Italian EQA scheme for haemocytometry, organized by the Istituto Superiore di Sanità, has been active for about five years (1984-1989). The aims of this programme were to evaluate the state of the art and to introduce in Italy a scheme recommended by ICSH. N. 126 public laboratories from different provinces joined voluntarily the programme and trials for haemoglobinometry (A01, A02), full blood count (B01-B08) and platelet count (D01-D03) were performed.

METHODS

Materials for testing consisted of blood lysate, preserved blood preparation containing native red cells and pseudoleukocytes, suspension of fixed platelets. The performances of laboratories was evaluated by consensus values (median, mean and standard deviation) and individual deviation index.

RESULTS

The instrument survey demonstrated that fully automated systems had the highest frequency. Non Gaussian distributions of results were often obtained and this was particularly true for WBC, PLT and MCV. The overall variability was lower than 5.5% for Hb, RBC and MCH and lower than 9.3% for other erythrocyte parameters; WBC and PLT counts displayed a higher dispersion (CV* = 9.8% and 25.4%); the spreading of results was strongly reduced in the homogeneous group of Coulter counters. In the course of the programme CV*s didn't show any relevant modification, a steady state performance being apparent. As regards the nature of variability, the random component was prevalent for all parameters, with the exception of MCV.

CONCLUSIONS

This pilot programme allowed to demonstrate the practicability of a national EQAS for haemocytometry according to the ICSH guidelines. Materials for testing showed acceptable stability, homogeneity and commutability. As regards analytical equipment as well as analytical variability, hospital centers participating in these EQA trials were comparable with laboratories taking part in similar EQAS of other European countries.

We evaluated the fully automated haematology analyser TOA Sysmex NE-8000 over a three-month period according to the ICSH protocol using as reference techniques a Coulter STKR counter and microscope examination for WBC differential and cell morphology. The NE-8000 employs aperture impedance to perform cell counts, with a sheath fluid to focus cells hydrodynamically prior to counting and sizing. WBC are differentiated into five populations. A combination of aperture impedance, radio frequency measurement and differential cell shrinkage is used, eosinophil and basophil percentages being established in two separate channels and subtracted from the total granulocyte count in order to give the value for neutrophils. Analysis of 1060 samples processed by the closed sampling automode demonstrated satisfactory counting performance. Among the WBC differentials obtained from 100 samples, neutrophil, eosinophil and lymphocyte results correlated well with those from microscopic examination of blood smears performed according to the NCCLS standard H20 T protocol. Differences observed in the percentages of basophils were of no biomedical significance. A comparative study for monocytes showed poor correlation for values below 5% and above 10%, best results being obtained in the intermediate range 5-10%. The NE-8000 also demonstrated good reliability for detection of abnormal cells.

The International Council for Standardization in Hematology(ICSH) is a non-governmental and non-profit body devoting to standardization work on hematology tests. It has publicized over 100 standardization documents on the subjects in the past 30 years. The ICSH is currently extending its activities to cooperative work with WHO, CEN, NCCLS, ICTH, ISBT, IFCC and ISLH. ICSH-Asia has also been founded to discuss local issues on standardization in hematology among Asian countries.

FSH (50 mu/rat X 12 days), ICSH (in the same dosis) and gonadotropinum sericum (20 U/rat X 13 days) have a prevailing inhibitory influence on the motor activity in untrained or/and trained rats. TSH raises the motor performance but significantly only in great doses (1.5 IU/100g body weight X 13 days). GH (0.1 and 0.5 mg/rat X 16 days) reduces the favourable effects of moderate training on the run performance. A part of these results could be explained by the studied biochemical changes.

Eleven monoclonal anti-IgG antibodies were evaluated by ten laboratories for possible AHG reagent use by the ISBT/ICSH protocol. Three of the anti-IgGs were selected by tests with weak Fya sensitized cells as this represents the best screening test. One of the IgM class anti-IgGs (205) equalled ISBT/ICSH reference reagents (R3P and RIIIM) against weak anti-D, -K and Fya sensitized cells and it reacted with all four subclasses of IgG. Thus, subject to satisfactory stability and extended trials with a wide range of weak antibodies, monoclonal anti-IgGs for AHG reagent use are now a possible alternative to conventional rabbit anti-IgG.

The BC-3200 automated hematology analyzer was evaluated and compared with the Beckman-Coulter AcT (Ac.T diff 2) 3-part differential hematology analyzer. The BC-3200 was evaluated according to guidelines published by the International Committee for Standardization in Hematology (ICSH), Clinical and Laboratory Standards Institute (CLSI), and Department of Food and Drug Administration (FDA). The results demonstrated no background, minimal carryover (<0.5%), and excellent linearity for hemoglobin (Hb) level, white blood cell (WBC), red blood cell (RBC), and platelet (PLT) counts (>0.998). Precision was generally acceptable for all complete blood count (CBC) parameters; coefficients of variation (CVs) were within the manufacturer's claims and CVs of CBC parameters, including WBC, RBC and PLT counts, Hb and mean corpuscular volume, were <6%. Correlation between the BC-3200 and Ac.T diff 2 was excellent (r>> 0.98) for all major CBC parameters (WBC, RBC, and PLT counts and Hb). We conclude that the overall performance of the BC-3200 is excellent and compares well with that of the Coulter Ac.T diff 2.

The Sysmex NE-8000 blood autoanalyser determines the values of red cells and platelets by impedance method and the differential leucocyte count (DLC) by means of radiofrequency plus two independent channels for eosinophils and basophils. The system is provided with a sampler capable of holding 100 closed tubes and its working velocity is about 120 samples per hour. The results of its evaluation are presented in this report. The parameters systematically provided by this cell counter include blood cell counts (plus haemoglobin rate, haematocrit and red cell indices), DLC platelet indices and volumetric distribution curves of red cells and platelets. The machine is provided with alarms on each of the above parameters for any suspicion of pathology. The accuracy analysis was performed with regard to the Technicon H*1 system and the conventional methods recommended by the ICSH. Precision, carry-over, linearity and effective velocity were evaluated in accordance to ICSH standards, and when assessing the DLC the number of visual revisions and the percentage of false positives and negatives were taken into account, and hence the sensitivity, specificity and efficiency of the machine. The accuracy of the parameters analysed was acceptable, except for MCHC and monocyte and eosinophil counts. A fair precision was found except for the monocyte count, along with excellent linearity except for high white-cell count. Less than 1.5% carry-over was observed, except for the leucocyte count. The number of necessary revisions of the DLC was somewhat higher than expected, the false negatives being below 11%. Sensitivity, specificity and efficiency were over 75%, except for hospital patients.(ABSTRACT TRUNCATED AT 250 WORDS)

The pituitary gland of the Herpestes edwardsi has been studies with special reference to the cytology of the pars anterior. In addition to the agranular chromophobic cells, six morphologically distinct chromophilic cell types have been observed in the pars anterior on the basis of selective histochemical and cytochemical staining techniques which can be classified into various groups such as STH, LTH, TSH, FSH, LH/ICSH and ACTH. The mucoid cells TSH, FSH and LH are much more concentrated in the centro-median region of the pars anterior. Highly concentrated colloid material appears in the acini or follicles which are scattered throughout the pars anterior. A relationship between the distribution of colloid material and any particular type of cell could not be established. The double intermedia layer surrounds almost completely the entire pars nervosa.

In the years 1984-1989 the Istituto Superiore di Sanità organized an EQAS for haematology (SVEQE) in Italy. A series of trials for haemocytometry, abnormal haemoglobins, HbA2, HbF, red cell G6PD and peripheral blood films, were carried out with the participation of 126 hospital laboratories, in different regions. SVEQE was an educative programme, aiming at promotion of quality assurance (QA) in laboratory haematology. At the same time an attempt was made to survey the analytical methods and instruments and to estimate the "state of the art" by the dispersion of all results. Participant laboratories were not scored for their performances. The operative protocol was harmonized to the guidelines established by WHO and ICSH; the trial specimens were prepared from normal or pathologic blood samples provided by blood banks or hospital departments. The trials for haemocytometry demonstrated a wide use of completely automated analyzers and in a steady state of performance during about five years. CVs, mainly for WBV and PLT, were somewhat higher than in other countries, where national QA systems have been established for a long time. Such discrepancies were not surprising in a pilot programme and were likely to be caused by inadequate internal quality control. The exercises for abnormal haemoglobins, HbA2, HbF and G6PD pointed out the need of using standardized methods according to the recommendations of ICSH. A large number of participating laboratories took part in the trial for blood cell morphology, being convinced of the educative function of this exercise; it is important to continue with systematic surveys, even including rare haematological disorders amongst the selected cases.

BACKGROUND

Monitor 100® (Electa Lab, Italy) is a newly developed automated method for measurement of erythrocyte sedimentation rate (ESR).

OBJECTIVE

The aim of our study was to compare the ESR values by Monitor 100® against the standard Westergren method.

METHODS

This cross-sectional study was conducted at a Level I trauma care center on 200 patients. The samples taken were as per the recommendations charted out by International Council for Standardization in Hematology (ICSH) for comparing automated and manual Westergrens method.

METHODS

Bland and Altman statistical analysis was applied for evaluating Monitor 100® against the conventional Westergren method.

RESULTS

The analysis revealed a low degree of agreement between the manual and automated methods especially for higher ESR values, mean difference -11.2 (95% limits of agreement, -46.3 to 23.9) and mean difference -13.4 (95% limits of agreement-58.9 to 32.1) for 1 and 2 hours, respectively. This discrepancy which is of clinical significance was less evident for ESR values in the normal range <25 mm/hour (-7.7 mean of difference; -18.9 to 3.5 limits of agreement).

CONCLUSIONS

The fully automated system Monitor 100® for ESR measurement tends to underestimate the manual ESR readings. Hence it is recommended that a correction factor be applied for the range of ESR values while using this equipment. Further studies and validation experiments would be required.