[Recommendations for metrological control of measuring equipments in the clinical laboratories]. / Recommandations pour la maîtrise métrologique des équipements de mesure au laboratoire d'analyses de biologie médicale (Document D).

The control of the measuring equipment (balances, mass standards, micropipettes, dilutors, volumetric glassware, thermometers...) is essential to obtain reliable analytical results. This control requires knowledge and use of the main standards, instructions for use, metrological verifications and confirmations as well as creation of checking and standardizing certificates ensuring metrological traceability. These items should help to control the quality of the analytical performances (fidelity and trueness in particular).

[Recommendations for expressing uncertainty of measurement of quantitative results in laboratory medicine]. / Recommandations relatives à l'expression de l'incertitude de mesure des résultats quantitatifs en biologie médicale (Document F).

In laboratory medicine, the quantitative results of examinations are interpreted with regard to reference intervals, clinical decision limits or previous results of a patient, from which it is necessary to inform the clinician about the uncertainty of measurement linked to the value of the result. This document explains the problematic of the expression of the uncertainty of measurement. It proposes recommendations concerning a simple way to evaluate uncertainty of measurement using long term internal quality control data and the value of the uncertainty linked to the method calibration. It approaches the determination of analytical goals and the choice of methods and also the comments accompanying the record of results and a help to their interpretation.

[Recommendations for the installation in the laboratory of metrological functions and corresponding documentation (Document B)]. / Recommandations pour l'installation dans le laboratoire de la fonction métrologie et de la documentation correspondante (Document B).

In order to ensure the quality of analytical results, clinical laboratories shall have a perfect control of process and equipments of measurement. Therefore is recommended individualisation of metrological function, generally entrusted the quality manager. This quality manager will draw up a metrological traceability, verifications and confirmations, control of non conformities, follow-up and evaluation of metrological function.

[Presentation for the use of a clinical laboratories of standards of metrology (Document A)]. / Présentation, à l'usage des laboratoires d'analyses de biologie médicale, des normes de métrologie (Document A).

Clinical laboratories shall have a perfect control of "in vitro diagnostic medical devices" to ensure the quality of their analytical results. This introductory presentation to a serie of documents of recommendations tackles the different standards of metrology concerning the requirements to reference materials, metrological traceability, metrological confirmation, management of measurement as well as uncertainty of measurement. The standards concerning clinical laboratories are then succinctly described.

Influence of handling procedures on rat plasma creatine kinase activity.

Ten-week-old Sprague-Dawley rats were held behind the front legs before and during anesthetic injection of sodium pentobarbital (group 1, 12 rats), or lifted by the base of the tail before and during injection (group 2, 12 rats). Creatine kinase (CK) values were higher (P = 0.004) in group 1 (median 564 IU/L) than in group 2 (median 272 IU/L). Handling rats by holding them behind the front legs may reduce the usefulness of CK activity as a measure of muscular disorders.

Evaluation of the Menarini-Arkray HA 8140 hemoglobin A1c analyzer.

We describe a multinational evaluation of the Menarini-Arkray HA 8140 hemoglobin (Hb) A1c analyzer, which utilizes a high degree of automation, including bar code reading, cap piercing, and whole-blood sampling. With-in- and between-batch CVs were < 2%. Linearity was confirmed throughout the working range of the analyzer. Common Hb variants, including Hb S, Hb C, and Hb F, did not interfere with the Hb A1c separation, and the potentially interfering labile Schiff base was effectively removed during the chromatographic procedure. The HA 8140 analyzer displayed good correlation to the Bio-Rad Variant analyzer, Tinaquant immunoassay, affinity chromatography, and an optimized "in-house" HPLC Hb A1c method. The methods when compared by Altman and Bland plots showed bias (upper, lower 95% confidence limits) of: Variant minus HA 8140 = 0.99 (0.23, 1.74), Tinaquant minus HA 8140 = 0.14 (-0.71, 0.98); affinity minus HA 8140 (after log transformation) = 1.13 (0.90, 1.41), and "in house" HPLC minus HA 8140 (after log transformation) = 0.91 (0.82, 1.01).

Geographic distribution of 119 alleles of the alpha and beta globin genes detected in 432 French Caucasian carriers of haemoglobin variant.

The French population is the result of mixing of different peoples including the Celts, Saxons, Germans, Italians and Hispanics. Between 1981 and 1993 patients were selected during investigations in France for haematological disorders associated or otherwise with the presence of a haemoglobin (Hb) variant. Further carriers of abnormal Hb were identified by HPLC measurement of glycated Hb in diabetics and by neonatal screening. Four-hundred and thirty-two subjects were found to be heterozygous for one of the 119 different alpha and the beta gene alleles encountered. These variants were characterised by a combination of 6 electrophoretic methods and in some cases by protein structure determinations. Some mutants reflected the population movements in and into France. A few mutants are frequently described in the French Caucasian population: Hb Lepore Boston, Hb D Punjab, whereas others appear to be anthropological markers. Hb Winnipeg has only been found in the West of France (Normandy); Hb J Baltimore is mainly found in French subjects of Spanish origin. Several cases of sporadic and previously undescribed mutations of Hb were identified. The last immigration waves from Africa and Asia appear to have contributed to the evolution of the pattern of haemoglobinopathies in the French population (Hb S, Hb O Arab or Hb C).

European multicentre evaluation of the ABBOTT Spectrum clinical chemistry analyzer.

The analytical performance of the selective multitest ABBOTT Spectrum analyser was studied according to the ECCLS guidelines and partly the CERMAB protocol in a multicentre evaluation involving laboratories from six European countries. Fifteen analytes, including the electrolytes sodium, potassium and chloride, were measured each in at least 3 laboratories, all at 37 degrees C, except the electrolytes, which are measured at room temperature. The trial lasted approximately three months and involved the collection of over 60,000 data points. It yielded the following results: 1. The precision was at least as good as the precision obtained with the comparison instruments. The majority of the coefficients of variation were between 1 and 4%. 2. The recovery for method assigned control sera values was, with few exceptions, within 10%. 3. Good agreement with respect to the method assigned values of control materials and method comparison with patient specimens to different instruments (e.g. SMAC, Hitachi 737, RA 1000) was found. 4. No drift was observed. 5. Reagent-related carry-over was not found. Specimen-related carry-over was detected in some cases, the deviation being of little or no clinical significance. 6. The manufacturer's claims regarding method linearity were as stated or exceeded. 7. The open system capability was tested and rated as very convenient. 8. The practicability of the instrument was very good.

Automated determination of urinary Na+, K+, chloride, inorganic phosphate, urea, and creatinine without sample dilution, with the "RA-XT".

We describe how concentrations of chloride, urea, inorganic phosphate, and creatinine in urine can be measured directly, without manual sample dilution, in a discrete analyzer (the Technicon "RA-XT"). These methods were accurate for concentrations of chloride up to 280 mmol/L, urea up to 500 mmol/L, inorganic phosphate up to 50 mmol/L, and creatinine up to 30 mmol/L. CVs are less than 3% nd results correlate well with those obtained by continuous-flow analysis (SMA-II). All these reagents are stable at room temperature for three weeks. Analyses are easy to perform and infrequent calibration is required.

Venocclusive disease of the liver after allogeneic bone marrow transplantation in man.

One hundred and fifty-one consecutive patients underwent allogeneic bone marrow transplantation (B.M.T.) following high-dose chemotherapy and single dose total body irradiation (T.B.I.) for hematologic malignancies between September 1980 and December 1985. All patients included in this study were treated using a 60 Co beam to deliver a prescribed dose of 10 Gy to the mid-plane of the abdomen. Total body irradiation was performed the day before B.M.T. The mean instantaneous dose-rate was 3.5 cGy/min (range: 2.6 to 4.7). The real dose received was measured using thermoluminescent dosimeters (lithium borate). The difference between the doses delivered to the liver and to the mid-plane of the abdomen did not exceed 5%. The mean real dose delivered to the reference point was 10 Gy (range 8.3 to 11.7). Ninety five per cent of the patients received a dose ranging from 9.1 Gy to 10.9 Gy. High-dose cyclophosphamide was given to 126 patients with a "standard-risk" of relapse (60 mg/kg on day 5 and 4 before B.M.T.). Chemotherapy was intensified by the addition of other drugs in 25 patients with "higher-risk" of relapse. We analyzed the effect of the following pretransplant characteristics on the subsequent posttransplant development of V.O.D.: age, sex, ASAT and/or ALAT before conditioning regimen, diagnosis and status of malignant disease, history of liver disease, interval between diagnosis of hematologic malignancy and B.M.T., conditioning regimen (i.e., classical or intensified) and dose delivered to the liver during T.B.I. Seventeen patients were classified as having clinical V.O.D. giving a prevalence of 11.2%. In the first 2 months following B.M.T., death occurred respectively in 9 of 17 (53%) and 23 of 134 (17%) patients with and without clinical V.O.D. Univariate analysis showed that four characteristics were significantly related to an increased prevalence of V.O.D.: sex (11/62 females vs 6/89 males; p less than 0.05); history of liver disease (7/28 vs 10/117 patients without antecedent; p less than 0.01); ASAT and/or ALAT levels greater than 1.5 upper normal limit (11/49 vs 6/102 patients with levels less than 1.5; p less than 0.01) and intensified conditioning regimen (6/25 vs 11/126 patients with classical regimen; p less than 0.05). The conditioning regimen and history of liver disease were highly correlated to transaminases levels. Only two factors, transaminases levels and female sex, remained significantly associated with V.O.D. after multivariate analysis.

Interaction of hemoglobin Siriraj with hemoglobin S: a mild sickle cell syndrome.

Hb Siriraj is a beta chain variant in which beta 7 (A4) Glu is replaced by a lysine. It has been encountered in association with Hb S in a black man from Martinique. Some properties of Hb Siriraj are compared, particularly, with Hb C [alpha 2 beta 26(A3)Glu----Lys], and a study of its in vitro interaction with Hb S is discussed.

Structural study of hemoglobin Hazebrouck, beta 38(C4)Thr----Pro. A new abnormal hemoglobin with instability and low oxygen affinity.

A new beta-variant has been detected and structurally defined in a French male, with a life-long history of hemolytic anemia. This variant is moderately unstable and has a low oxygen affinity. The abnormal hemoglobin was not detected by standard electrophoretic procedures. It moved slightly slower than Hb A during isoelectric focusing (IEF). Two minor fractions were also seen; the first migrated just cathodal to Hb F, as did partially oxidized Hb A or hemichrome derivatives of some unstable hemoglobins; the second in the position of free alpha-chains. The abnormal beta-chain was readily separated from both beta A- and alpha A-chains by acid-urea-Triton globin chain electrophoresis. Structural study was conducted simultaneously by fingerprinting and high-performance liquid chromatography (HPLC) of tryptic peptides. A new mutation beta 38(C4)Thr----Pro was found, which was named Hb Hazebrouck.

A new silent hemoglobin variant in a black family from French West Indies, hemoglobin Le Lamentin alpha 20 His replaced by Gln.

A new abnormal hemoglobin Hb Le Lamentin alpha 20 (B1) His replaced by Gln was discovered during a survey of cord blood from the French West Indies (Martinique). This variant displays an electrophoretic pattern similar to that of Hb A but can be isolated by isoelectric focusing (IEF) and Biorex 70 chromatography. Family studies showed the presence of this hemoglobin variant in the father and in two of his three children. Hematological data from the carriers were normal.

An update on electrophoretic and chromatographic methods in the diagnosis of hemoglobinopathies.

This review primarily deals with methods for separations of hemoglobins. An introduction considers electrophoretic methods as well as those involving isoelectric focusing and chromatography. The main advantages or disadvantages of each procedure are discussed after each technical description. The chromatographic methods are mainly limited to those used in clinical biochemistry. The second section treats the main diagnostic problems typically met with in the field of the hemoglobinopathies and deals successively with the diagnosis of hemoglobinopathies in the adult and the newborn. Numerous variants have been described in the adult, and among them Hb-S and Hb-C variants are the most frequent. Unstable or high oxygen affinity variants of hemoglobin are also considered. Finally, a new strategy for diagnosis is proposed. A special section is devoted to the diagnosis of thalassemia syndromes. The prenatal diagnosis of hemoglobinopathies is also discussed in some detail with a view to preventing the birth of homozygous children. This update ends with a chapter on the interest of the assay of hemoglobins A1c in the pathology of diabetes mellitus.

[Detection of haemoglobinopathies at birth, using isoelectric focalization (author's transl)]. / Dépistage des hémolgobinopathies à la naissance par isoélectrofocalisation. Etude de la population de la martinique.

The incidence and nature of haemoglobinopathies were investigated at birth in Martinique, where 4635 samples of umbilical cord blood were examined. Conventional blood values were determined, and a new, extremely simple and highly selective test was performed: isoelectric focalization. Abnormalities were found in 14.3% of blood samples, viz: A/S 7.25%, A/C 3.17%, S/S 0.17%, S/C 0.24%, C/C 0.04%, other mutations 0.63% alpha-thalassaemia minor (alpha 1-thal) 1.72% and isolated microcytosis 1.01%. Unusual abnormalities included 7 different mutations of the 4a-chain, 3 of the beta-chain and 3 of the gamma-chain. The clinical and epidemiological importance of such investigations is emphasized.

[Clinical studies of the erythrocytosis produced by a new hemoglobin with oxygen affinity : Hb Hôtel-Dieu alpha 2A beta 2 Asp replaced by Gly (author's transl)]. / Polyglobulie par hémoglobine anormale hyperaffine pour l'oxygène : l'hémoglobine Hôtel-Dieu alpha 2 A beta 2 99 Asp replaced by Gly.

A new case of erythrocytosis associated with a high oxygen affinity hemoglobin is reported : the substitution of beta 99 of aspartic acid by glycine, characterized this abnormal hemoglobin, named "hemoglobin Hôtel-Dieu". An important associated smoking habit, probably responsible of an abnormal lung X-ray and of a disturbed CO transport, contributed to increase the difficulty of diagnosis. Familial inquiry, the search of an abnormal hemoglobin by standard or isoelectrofocusing electrophoresis, and the 2,3-DPG assay should be systematically performed in case of polycythemia in young people or in the absence of obvious aetiology. A corpuscular abnormality can be thus detected in about 10 p. cent of such polycythemias.

Hemoglobin Hotel-Dieu beta 99 Asp replaced by Gly (g1). A new abnormal hemoglobin with high oxygen affinity.

Hemoglobin Hotel-Dieu was detected by isoelectric focusing during investigation of a patient who had erythrocytosis. This variant migrates on cellulose acetate electrophoresis to a cathodic position relative to Hb F. In hemoglobin Hotel-Dieu, aspartic acid is substituted by glycine in position 99 of the beta chain. As in other abnormal hemoglobins in which substitution of this residue has occurred, Hb Hotel-Dieu exhibits a high oxygen affinity and is associated with familial erythrocytosis.