We performed a parallel evaluation of 5 automated reticulocyte analyzers. The guidelines were those proposed by the National Committee for Clinical Laboratory Standards and the International Council for Standardisation in Haematology. Duplicate analyses were performed for 225 healthy subjects and 115 patients affected by various diseases. The reference intervals were different for each method (ADVIA 120, 27-125 x 10(3)/microL [27-125 x 10(9)/L]; CELL DYN 4000, 25-108 x 10(3)/microL [25-108 x 10(9)/L]; GEN-S, 20-85 x 10(3)/microL [20-85 x 10(9)/L]; SE 9500 RET, 23-95 x 10(3)/microL [23-95 x 10(9)/L]; and VEGA RETIC, 30-130 x 10(3)/microL [30-130 x 10(9)/L]). The comparisons of percentage counts with the microscopic reference method were satisfactory for all automated methods. However, a tendency to overestimate at low counts was noted. This progressively increased from the SE 9500 RET to the VEGA RETIC. The imprecision was excellent for all the methods at normal and high concentrations. This was higher at low concentrations. When compared with the microscopic reference, the analyzers showed satisfactory sensitivity at low counts and excellent sensitivity at high counts. The overall agreement varied from 74.8% for the GEN-S to 86.1% for the SE 9500 RET.

Erythrocyte histograms should be analysed in accordance with the ICSH recommended protocol to determine: How many red cell populations are present and the proportion of the total red cells which are included in each population. The central tendency (mode, median & mean) and dispersion (size ratio, geometric standard deviation & coefficient of variation) for each population. The proportion of microcytic or macrocytic cells in each population. Examples are given to show how the patient's diagnosis is assisted by careful analysis of the histograms and the effect of treatment monitored by sequential testing. Histograms can be generated from blood films or from aperture-impedance and light-scatter volume measurements. Volume measurements on these systems are influenced by the red cell shape and internal refractive index. Mean cell hemoglobin concentration affects both shape in the aperture-impedance orifice and internal refractive index in light-scatter systems. There is poor agreement between volume histograms obtained on the two measuring systems and the histograms available on current automated instruments provide no more useful information than can be obtained from the blood film. Automated instruments need to produce histograms with fewer artefacts and the histograms should then be examined in accordance with the ICSH protocol. This approach should maximise the diagnostic value of the complete blood count.

Rabbit antisera to rat prolactin was found to cross-react with the hamster pituitary extract by double diffusion and quantitative precipitin techniques. Short term neutralization of prolactin by antiserum to rat prolactin either in early or late stages of gestation did not have any deleterious effects. Prolonged administration of rat prolactin antiserum resulted in an increase in the incidence of resorptions, but pregnancy was still maintained followed by normal parturition. This is in marked contrast to the abortive effects of an antiserum of ICSH. It was concluded that prolactin does not play a major role in maintaining pregnancy of the hamster.

The technique recommended by the International committee for Standardization in Haematology (ICSH) was utilized for determination of erythrocyte sedimentation rate (ESR) in 'healthy' young Nigerian students living in the northern part of the country. Ninety-five point eight percent of a sample of 188 cases showed values within the Westergren norm for the first hour viz. 2.6 mm for males and 8.4 mm for females. A detailed follow up during the second hour revealed that 80% were within the norm for the second hour. The second hour elevations in apparently healthy subjects never exceeded 42 mm into the pathological range which would invalidate its use as a screening test in Nigeria for a young student population. The effects of standardization of technique and the changing age-nutrition parameters of populations on the phenomenon of erythrocyte sedimentation is evaluated in the context of the view that the 'African normal' is higher than in temperate countries.

The theory of reference ranges advocated by the International Committee for Standardization in Haematology (ICSH) was tested experimentally for an aperture impedance and a flow cytometric system. The reference sample was drawn from volunteer blood donors and the sampling procedure carefully standardised. Subjects were partitioned for gender and smoking habits, and their ages noted. Reference ranges were constructed from Gaussian statistics, after objective evaluation of the data, based on an assumed occurrence of one false positive measurement in every twenty samples. Different reference ranges were obtained on the two different systems for some haematological parameters, and partitioning also produced differences in calculated reference ranges. We conclude that strict adherence to the ICSH recommendations is almost impossible. Nevertheless, it is important that all laboratories should construct their own local reference ranges for their specific analytical systems before cost effective clinical decisions can be made.

Haemoglobin A2 levels were assessed using a modular high performance liquid chromatography (HPLC) system with a protocol designed for the measurement of haemoglobin A1C. There was good correlation (r = 0.96; P < 0.001) between this technique and the International Committee for Standardisation in Haematology (ICSH) recommended method of microchromatography. The range of haemoglobin A2 was found to be 2.3-3.2% in apparently normal individuals, and 4.0-6.7% in those with beta-thalassaemia trait. The HPLC system produced reliable results quickly for haemoglobin A2 with no alteration to the protocol used for measuring haemoglobin A1C.

BACKGROUND

The BC-3600 Auto Hematology Analyzer (hereinafter call BC-3600) is a quantitative, automated hematology analyzer and leukocyte differential counter for In Vitro Diagnostic Use in clinical laboratories.

METHODS

The analyzer was evaluated and compared with the Mindray BC-3200 3-part differential (BC-3200) and Sysmex XE-2100 5-part differential (XE-2100) Hematology Analyzer in the hematology laboratory of a university hospital. The BC-3600 was evaluated according to guidelines published by Clinical and Laboratory Standards Institute (CLSI), the International Committee for Standardization in Hematology (ICSH), and Department of Food and Drug Administration (FDA).

RESULTS

There were no background, minimal carryover (<0.5%), and excellent linearity for white blood cell (WBC), hemoglobin (Hb) level, red blood cell (RBC), and platelet (PLT) counts (r>> 0.999). Precision was good at all levels for the routine cell blood count (CBC) parameters: CV% being ≤2.0, except for platelet count (PLT) at the low level with CV% of ≤5.0% and WBC at the low level with CV% of <3.0%. Correlation between the BC-3600 and BC-3200, XE-2100 were excellent (r>> 0.99) for all major CBC parameters.

CONCLUSIONS

It is concluded that the overall performance of the BC-3600 is excellent and compares well with that of BC-3200 and XE-2100.

In this study we evaluated the analytical performances of the Cell Dyn 3500, an automated hematology analyzer that provides the electronic and optical detection of leukocyte count and the complete cell counts in samples of whole blood. The protocol included evaluation of complete blood count and differential leukocyte count parameters, using the ICSH and the NCCLS H20-A protocols. Technical performances with regards, to linearity, carry over, precision and stability were quite well acceptable; the accuracy showed a good agreement between Cell Dyn 3500 and instruments used in our laboratory for the major hematologic indices and a good correlation for neutrophils, lymphocytes and eosinophils compared with the manual count.

The aim of this study was to compare the performance of the automatic TEST 1 ESR system, SIRE Analytical Systems (TEST 1), with that of the the Sedisystem 15, Becton Dickinson (SEDI), and the International Council for Standardization in Haematology reference method (Westergren) for measuring the length of sedimentation reaction in blood (LSRB). This reaction was measured in 418 paired blood samples drawn in K2-EDTA vacuum tubes and specific tubes from patients scheduled for routine LSRB measurement. The TEST 1 system uses micro-sedimentation and quantitative capillary photometry technology, whereas the SEDI uses a CCD camera. For Westergren, a 200 mm column with 3.0 mm internal diameter was used. Compared to Westergren, TEST 1 gives accurate values of LSRB in most of the samples (mean of differences: 0.99 +/- 10.4 mm; 95% CI, -0.807 to 2.78 mm; n =131). Similar results were obtained in the comparison with SEDI (mean of differences: -0.626 +/- 8 mm; 95% CI, -1.756 to 0.5 mm; n = 195). Compared to those of fresh blood samples, LSRB values were significantly lower in 24 h stored samples, either at 4 degrees C (21.5 +/- 2.3 vs. 19.4 +/- 2.2 mm; p (Spearman's coefficient of correlation): 0.981; n = 44) or at room temperature (19.1 +/- 2.5 vs. 16.2 +/- 2.1 mm; p: 0.903; n = 46). In conclusion, TEST 1 is a rapid, reliable system for automatic measurement of LSRB in standard K2-EDTA blood samples. It has a very low imprecision and maintains a good performance in 24 h stored samples. In addition, due to its operational characteristics (60 samples/20 min) it is a suitable tool for clinical laboratories with a high work load as well as for emergency laboratories.

One of the many challenges facing laboratories is the verification of their automated Complete Blood Count cell counters for the enumeration of body fluids. These analyzers offer improved accuracy, precision, and efficiency in performing the enumeration of cells compared with manual methods. A patterns of practice survey was distributed to laboratories that participate in proficiency testing in Ontario, Canada, the United States, the United Kingdom, and Japan to determine the number of laboratories that are testing body fluids on automated analyzers and the performance specifications that were performed. Based on the results of this questionnaire, an International Working Group for the Verification and Performance of Automated Cell Counters for Body Fluids was formed by the International Council for Standardization in Hematology (ICSH) to prepare a set of guidelines to help laboratories plan and execute the verification of their automated cell counters to provide accurate and reliable results for automated body fluid counts. These guidelines were discussed at the ICSH General Assemblies and reviewed by an international panel of experts to achieve further consensus.

A prototype of the CELL-DYN 3200 haematology analyser was evaluated in a tertiary care hospital laboratory. Precision, effects of sample ageing, linearity, carry-over, and comparability of cellular blood counts and five-part leucocyte differentiation were determined in accordance with the ICSH guidelines for the evaluation of blood cell analysers; the results were satisfactory for all parameters tested: haemoglobin concentration, RBC, MCV, WBC, platelet count, and counts of neutrophils, lymphocytes, monocytes, and eosinophils. Two-hundred and forty-seven routine blood samples were used for the comparability studies. The cellular blood count results from the CELL-DYN 3200 and the Bayer Diagnostic H-1 systems corresponded closely (correlation coefficient r>> 0.96 for all parameters). For 201 samples without an instrument-generated suspect flag the same was true with regard to the differential parameters, although somewhat lower correlation was observed for monocyte counts (r = 0.88). Comparisons to 400-cell microscopic differentials gave similar results (r>> 0.93 for neutrophil, lymphocyte and eosinophil counts). Our results suggest that the CELL-DYN 3200 analyser will serve the needs for automated blood cell counting and differential leucocyte counting in a tertiary care hospital laboratory.

The erythrocyte sedimentation rate (ESR), now more appropriately referred to as the "length of sedimentation reaction in blood (LSRB)", remains the most widely used laboratory test for monitoring the course of infections, inflammatory diseases and some types of cancer. Thanks to the several recently developed methods for the measurement of this reaction, the safety and reliability of LSRB testing procedures have improved. The method for LSRB measurement recommended by the International Council for Standardization in Hematology (ICSH) and the National Committee for Clinical Laboratory Standards (NCCLS) is based on the traditional Westergren method, using EDTA-anticoagulated samples. The present paper describes and evaluates a procedure for LSRB measurement with a new manual system, the Microtest 1, which requires only 30 microl of blood and is optimal for pediatric use. Microtest 1 results correlated satisfactorily with the ICSH recommended method (r=0.88, 95% CI 0.84-0.91; y=3.45+0.85x), had no significant bias (2.15, 95% CI -0.32-4.63) and had an imprecision of less than 7% for the reference range values. In addition, the results obtained with the Microtest 1 in "native" whole blood were comparable with those obtained with the reference method in K3EDTA and sodium citrate-anticoagulated specimens; (bias = 1.19, 95% CI -1.78 to 4.16 for K3EDTA-anticoagulated samples and bias = 2.43, 95% CI -0.58 to 5.44 for sodium citrate-anticoagulated samples).

The Fifth International Society of Blood Transfusion (ISBT) Platelet Serology Workshop took place in 1990. A total of 21 laboratories participated in the wet workshop. Results were discussed at a meeting of the International Committee on Standardization in Haematology (ICSH) Expert Panel on Platelet Serology at the Joint Congress of the ISBT and the American Association of Blood Banks in Los Angeles in November 1990. Each of the participants analyzed 37 serum samples for platelet-reactive alloantibodies and isoantibodies. In addition, two drug-dependent (quinine) platelet-specific antibodies had to be identified among a subset of seven sera. For the first time in a platelet serology workshop, five platelet suspensions had to be typed for the alloantigens of the HPA-1, -3, -4, and -5 systems.

We report a potentiometric fully automated method for determining red cell glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase activities and the glucose 6-phosphate dehydrogenase/6-phosphogluconate dehydrogenase index using 25 microliters of whole blood. No sample pre-treatment (e.g., preparation of the haemolysate) is needed and the measurements are performed at pH 8.0 and 37 degrees C under the conditions recommended by the ICSH committee. The reproducibility was constantly good, with within-run CV of 1.0% (glucose 6-phosphate dehydrogenase) and 5.9% (glucose 6-phosphate dehydrogenase/6-phosphogluconate dehydrogenase) for activities in glucose 6-phosphate dehydrogenase non-deficient adults, and of 2.3% (glucose 6-phosphate dehydrogenase, G6PD) and 2.5% (glucose 6-phosphate dehydrogenase/6-phosphogluconate dehydrogenase) for G6PDMediterranean heterozygotes. Linearity was observed up to an activity of 2800 U/l of glucose 6-phosphate dehydrogenase. Results of glucose 6-phosphate dehydrogenase activity (U/l) in whole blood (y) correlated well with those obtained with the previously described monostarter assay, performed at pH 9.2 (y = 0.60x + 37; n = 80; r = 0.991). Results of 6-phosphogluconate dehydrogenase (U/l) in whole blood (y) correlated well with those obtained by the ICSH recommended method (x) (y = 0.779x - 44; n = 23; r = 0.991). Reference intervals are reported for glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase and glucose 6-phosphate dehydrogenase/6-phosphogluconate dehydrogenase index relatively to normal, beta- and alpha-thalassaemic glucose 6-phosphate dehydrogenase non-deficient adults, to glucose 6-phosphate dehydrogenase deficient adult males and to G6PDMediterranean non-thalassaemic heterozygotes. We demonstrate that the diagnostic sensitivity of the glucose 6-phosphate dehydrogenase/6-phosphogluconate dehydrogenase index in detecting the G6PDMediterranean heterozygotes is superior to that of the glucose 6-phosphate dehydrogenase activity alone.

Since the first systematic blood volume studies of polycythemia in the 1920s, measurement of blood volume and red cell mass (RCM) has become routine. However, the radionuclide-labeling methods promulgated by the International Committee for Standardization in Haematology (ICSH) remain complex and poorly understood. Many hematologists and other clinicians err in the belief that these methods permit "direct measurement" of RCM, whereas the ICSH method is indirect: it requires calculation of RCM from (PCV) X (whole blood volume). The use of an elevated value of PCV to calculate RCM in order to evaluate the same elevated value of PCV is a curiously circular logic that is embraced by most clinicians and most hematologists. Analysis of published data in 186 cases of polycythemia vera indicates that RCM is an exponential function of PCV. In most cases, PCV alone suffices to document normal or increased RCM. Relative polycythemia results from dehydration, not from stress. Clinicians need to be aware of the range of physiologic fluctuations that normally occur in plasma volume. Realistic criteria for normal ranges of PCV, Hb concentration and RCM should be adopted in clinical laboratories so that clinicians will not be misled to undertake futile and costly investigations of results that are in the upper percentiles of the normal distribution, as exemplified by the Ulysses Syndrome.

Since the first systematic blood volume studies of polycythemia in the 1920s, measurement of blood volume and red cell mass (RCM) has become routine. However, the radionuclide-labeling methods promulgated by the International Committee for Standardization in Haematology (ICSH) remain complex and poorly understood. Many hematologists and other clinicians err in the belief that these methods permit "direct measurement" of RCM, whereas the ICSH method is indirect: it requires calculation of RCM from (PCV) x (whole blood volume). The use of an elevated value of PCV to calculate RCM in order to evaluate the same elevated value of PCV is a curiously circular logic that is embraced by most clinicians and most hematologists. Analysis of published data in 186 cases of polycythemia vera indicates that RCM is an exponential function of PCV. In most cases, PCV alone suffices to document normal or increased RCM. Relative polycythemia results from dehydration, not from stress. Clinicians need to be aware of the range of physiologic fluctuations that normally occur in plasma volume. Realistic criteria for normal ranges of PCV, Hb concentration and RCM should be adopted in clinical laboratories so that clinicians will not be misled to undertake futile and costly investigations of results that are in the upper percentiles of the normal distribution, as exemplified by the Ulysses Syndrome.

The correct enumeration of platelets is still an elusive matter. This is mainly due to the fact that commercial instruments which are used for platelet counting cannot discriminate platelets from other cellular particles and precipitates that cause similar signals. Visual (chamber counting) methods are still frequently used in routine laboratories to verify low automated platelet counts (< 50 x 10/l) despite obvious technical and statistical drawbacks. The following report shows how platelet counts can be measured by multiparameter flow cytometry with the help of reference particles (fluorescent latex beads) and platelet-specific antibodies i.e. anti-GPIIb/IIIa(CD41a), anti-GP Ib-alpha (CD42b) and anti-GP IIIa (CD61). The linearity of this method was highly satisfactory and the observed imprecision was within acceptable limits. At a platelet concentration of 10 x 10(9)/l the coefficient of variation (CV, n = 10) ranged from 5.3% (PCV = 0.456) to 5.6% (PCV = 0.148). Accuracy was evaluated by comparing results to the ICSH-selected method for platelet counting. The correlation of both methods was significant (P < 0.005) and Passing-Bablok's linear regression analysis showed no systematic differences between the two methods. Comparisons of this new platelet counting technique were also performed with routine visual methods, automated blood analysers (Technicon H-1, Sysmex E-5000) and a different flow cytometric method using only forward and side light scatter properties of platelets for their discrimination. The linear correlation of all methods was significant (P < 0.01) at platelet concentrations above 50 x 10(9)/l. At lower platelet concentrations, our new platelet counting technique correlated significantly only with the visual and the forward/side scatter methods.(ABSTRACT TRUNCATED AT 250 WORDS)