[Immunohematological surveillance of the pregnant woman: new prevention policy]. / Suivi immunohématologique des femmes enceintes: nouvelles recommandations.

Despite the generalization of immunoprophylaxis by anti-RH immunoglobulins over 40 years, fetomaternal incompatibility due to RH1 antigen (RhD) is not completely eradicated, although perinatal consequences might be extremely serious. Additionally, allo-immunizations against other antigens, especially anti-RH4 (anti-c) and anti-KEL1 (anti-Kell), may cause severe haemolytic disease. Follow-up of allo-immunization during pregnancy and its prevention are therefore still a concern for all pregnant women. Immunohaematological tests used in antenatal patients are under practice for a long time. However, despite significant progress, it is clear that these tests provide only an indirect indication and will only help the obstetrician, in conjunction with over fetal parameters, to assess the severity of the haemolytic disease. Since almost two decades, fetal RHD genotyping became a reality, first using amniocytes, but more recently by analyzing fetal DNA present in the maternal plasma. RH prophylaxis concerns RH:-1 women, who are non-sensitized against RH1 antigen during and at the end of their pregnancy with a RH1 child. RH prophylaxis includes targeted prophylaxis after foetomaternal haemorrhage and now routine antenatal RH prophylaxis at 28 gestation weeks. Indications for RH prophylaxis and immunohaematological testing to assure an efficient therapeutic prevention have been summarized in France through specific recommendations of the National College of Gynecologists and Obstetricians.

[ABO-RH1 grouping and red blood cell antibody screening: error analysis in the French external quality assessment in 2006]. / Groupes sanguins et recherche des anticorps antiérythrocytaires: analyse d'erreurs au contrôle national de qualité en 2006.

Further to a survey set up in 2006 over 2600 laboratories by the French agency for health product safety (AFSSAPS), seven ABO grouping errors and 53 negative answers to red blood cell antibody screening with a serum containing anti-RH1 antibody, have been found. A questionnaire sent to the involved laboratories revealed non-analytical errors already met in previous cases. Besides, the analytical stage has also induced red blood cell antibody screening errors due to a wrong serum collection by the automat. Here are displayed the analysis of the questionnaire results and proposed corrections.

[Red blood cell antibody screening: error analysis in a french interlaboratory comparison program survey]. / Recherche des anticorps antiérythrocytaires: analyse d'erreurs au Contrôle national de qualité

The French quality control is organized by the French Health Products Safety Agency. In 2005, the immuno-haematology testing control included the screening of an anti KEL 1 antibody. 17 out of 2639 laboratories (0,64%) answered 'negative screening'. All laboratories received a questionnaire in order to understand the failure. In this paper the authors present the detailed laboratories' responses and failure explanations.

[Validation of antibody screening by indirect antigloblin test and ABO blood typing by filtration and microplate techniques: assessment of robustness]. / Maîtrise des tolérances métrologiques : application à la recherche des anticorps anti-érythrocytaires (RAI) en test indirect à l'antiglobuline (TIA) et au groupage sanguin ABO par les procédés de filtration et en microplaque.

According to requirements of the French Committee for Accreditation (Cofrac), it is essential to use validated and standardised methods in Immunohematology. This imposes first the knowledge of metrological tolerances for all the technics. Two multicenter studies were carried out to define the maximal acceptable deviations concerning incubation temperature and time, volumes of patient plasma and tests cells for antibody screening using indirect antiglobulin test on one hand and for reverse grouping on another hand. All equipment used (temperature test chamber, chronometer, pipettes) were calibrated according to Cofrac standards. The antibody screenings were performed manually using 3 different filtration systems: ID Diamed, Biovue Ortho and Scangel Biorad, the same tests cells, a standard 20 ng/mL anti RH1, a positive control anti KEL1 and a negative control; the reverse blood grouping was performed manually using the above mentionned filtration systems and microplate technic with the same A1 and B test cells. These two studies showed that all the tests from the multiples combinations of the above parameters gave the same results and allowed us to define a range of tolerance for 4 critical physical parameters involved in the antibody screening and blood typing.

[Quality control: sensitivity of the indirect antiglobulin test by filtration]. / Evaluation externe de la qualité: étude de la sensibilité du procédé de filtration en test indirect à l'antiglobuline.

For ten years, the working party of immunohaematology of the French Society of Blood Transfusion organizes a quality control. After the modification of the law about the realization of erythrocyte typing and detection of unexpected red cell allo-antibodies, the quality control was performed in order to determine the sensitivity of the indirect antiglobulin test by filtration with a standard anti-RH1(D) produces by the National Reference Center of Blood Groups.

[Critical analysis of the new ministerial order about guidelines in immunohaematology]. / Nouvel arrêté de bonnes pratiques d'immunohématologie: analyse critique.

The evolution of the methods used in immunohematology is a constant concern for all those involved in the process. Thus, during the last few years, new technologies have been introduced which aim at improving performance. This improvement is not limited to the search for an overall increase in specificity-sensibility; it also takes into account the capability to detect "the clinical significant" as well as the limitations of human reliability, which justifies the introduction of automation and computerization. The whole of these methodological evolutions associated with that of the performance of re-agents, legitimate to modify the law about the realization of erythrocyte typing and detection of irregular antibodies. In order to ensure the immunohematologic safety of transfusions, it was also necessary to place the entire transfusion process under computer control: patient identification, sample collection, test results and release of blood.

[Follow-up of the feto-maternal allo-immunization]. / Suivi de l'allo-immunisation foeto-maternelle.

Immunohaematological tests used in antenatal patients have come a long way. However, despite a great deal of progress, we should not loose sight of the fact that these tests give only an indirect measurement and will only help the obstetrician, in conjunction with other fetal parameters, to assess the severity of the haemolytic disease (HD) of the fetus and newborn. The best method to assess the severity is the direct determination of foetal blood group hemoglobin after foetal blood sampling but this procedure is not without risk. Since 10 years ago, it is possible to determine the RHD genotype of the fetus using amniocytes and, today, maternal plasma directly. All pregnant women should be grouped for ABO-RH-KEL1 and the sera tested for clinically irregular antibodies (anti-RH are still the most frequent). The trend in anti-RH levels is more important than the level itself. The perfect technique for anti-RH quantitation has not been developed. Manual titration is simple but only provides rough, semiquantitatives estimates of anti-RH concentration. Quantitative haemagglutination methods, using auto-analyzers and appropriate anti-RH1 standards, measure in microg/ml, are sensitive, rapid and have acceptable intra-laboratory reproductibility.

[External quality evaluation]. / Evaluation externe de la qualité.

Since ten years, the immunohaematology working group of the French Society of Blood Transfusion has organized a quality control. Tests concern essentially the screening and identification of irregular antibodies, direct antiglobulin tests and elutions.

[Problem-solving in immunohematology: direct compatibility laboratory test ]. / Aide à la décision en immunohématologie: épreuve directe de compatibilité au laboratoire (EDC).

Cross-matching between the serum of a patient and the red blood cells to be transfused is most important for the prevention of hemolytic transfusion reactions in allo-immunized or new-born patients found positive with direct antiglobulin test. Cross-matching is a time-consuming and complex laboratory test. In order to obtain valid results, it is necessary to abide by some technical rules detailed in this article. The choice of the blood units to be cross-matched depends on the patient's clinical story and on the specificity of anti-erythrocyte antibodies present in the serum. The identification and the management of most frequent difficulties met by using the cross-match technique are discussed hereby.

Decision aides in immunohematology: detection of anti-erythrocyte antibodies. / Aide à la décision en immunohématologie: recherche des anticorps anti-érythrocytaires (RAI).

Detection and identification of irregular red-cell antibody in the serum or plasma of a patient is of prime importance for the prevention of hemolytic transfusion reactions and the biological supervision of the hemolytic disease of the foetus or the newborn. Practice in these tests is replete with complex biological problems. Using problem solving strategies, we discuss the recognition and resolution of the most frequent difficulties encountered in red cell antibody identification.

[Evolving methods in immunohematology]. / Evolution des méthodes en immunohématologie.

The immunogenic nature of erythrocyte polymorphism is in variance with the incompatible transfusion. Indeed, the fixing of an antibody on the corresponding antigen generally condemns the cell concerned with its destruction. Therefore, in order to ensure the immunohemolytic safety of the transfusions, it is necessary to avoid an in vivo encounter between antigens and antibodies, whose feasibility study in vitro is a determining element. Because of the requirement standards of such analyses and the preoccupation with the continuous improvement of transfusion safety, the evolution of the methods used in immunohematology is a constant concern for all those involved in the process. Thus, during the last few years, new technologies have been introduced which aim at improving performance and sometimes implementing alternatives to agglutination. This improvement is not limited to the search for an overall increase in specificity-sensitivity; it also takes into account the capability to detect "the clinically significant" as well as the limitations of human reliability, which justifies the introduction of automation and computerization. The whole of these methodological evolutions associated with that of the performance of reagents, legitimate the need to reconsider the realization of erythrocyte typing and the search for anti-erythrocyte antibodies.

[Analytical validation in erythrocyte immunohematology]. / Validation analytique en immunohématologie érythrocytaire.

In a transfusional or foeto-maternal context, hemolysis by incompatibility due to anti-erythrocyte antibodies (regular or irregular) remains the most frequent and most serious immunological risk in the receiver. In order to prevent this risk, a number of actions must be taken, such as the realization of the immunohematologic analyses for which the methodological practices have been legislated because of their serious clinical consequences. Several elements play a role in the reliability of the analyses and their results: the selection of the reagents and their validation in the routine technique used; the validation of reception; the controls involved in secondary preparations (e.g., blood cells reagent); and the daily internal controls. All this requires the choice of adapted controls and the management of possible anomalies.

[Problem-solving in immunohematology: interpretation of ABO typing and its difficulties]. / Aide à la décision en immunohématologie: interprétation du groupage sanguin ABO et de ses difficultés.

Practice in immunohematology is replete with complex problems that require practitioners' problem-solving performance. In immunohematology, the acquisition of the reasoning process and necessary skills for making clinical decisions is based on teaching problem-solving strategies which potentially reduce errors and improve patient outcome. We discuss the recognition and resolution of the common causes of discrepancies in ABO typing results using problem-solving strategies.

[Prevention of fetal hemolytic disease: it is time to take action]. / Prévention de la maladie hémolytique du foetus et du nouveau-né: il faut agir!

In spite of the progress made since 1970 in specific prevention by anti-rhesus immunoglobulins, and improved management of at-risk pregnancies, allo-immunization due to the erythrocytic Rh 1 antigen (formerly known as Rhesus D or Rh D) remains widespread. In fact, anti-Rh 1 antibodies currently constitute over one-third of the immune antibodies detected after pregnancy. The prevention of allo-immunization against the Rh 1 antigen is therefore still problematical, and concerns approximately one pregnant woman in seven. The etiology and pathology of fetal hemolytic disease have been recalled, and the treatment approach during pregnancy and delivery has been carefully examined. Tests for quantifying the risk of fetomaternal hemorrhage have also been described. This approach aims at improving the methods of preventing allo-immunization (e.g., during pregnancy and delivery) and the efficacy of treatment. It is also stated that if the necessary preventive action is not taken in cases of allo-immunization due to to the Rh 1 antigen, this should be considered a grave medical fault.

[Detection of irregular anti-erythrocyte antibodies using the indirect antiglobulin test in a low-ionic-strength medium. Immunohematology Group of the French Blood Transfusion Society]. / Recherche d'anticorps irréguliers anti-érythrocytaires (RAI) en test indirect à l'antiglobuline en milieu de basse force ionique. Groupe Immunohématologie de la Société Française de Transfusion Sanguine.

The detection of irregular antibodies is usually performed with serum by the indirect antiglobulin test (IAT) with a polyspecific antiglobulin reagent. In a first study in 1996, we compared the results obtained with 3,264 blood samples of patients drawn with or without anticoagulant: no significant difference was observed among the 240 allo-antibodies detected and identified. In this paper we report the comparison of the results obtained by IAT on column of filtration with two kinds of reagents: polyspecific and anti-IgG antibodies. Respectively 2,927 (76 contained an antibody), and 643 (161 contained an antibody) sera of patients were tested with methods ID-Diamed and Ortho-Biovue. Titrations of 153 other antibodies were also performed with the two reagents. Results showed no significant difference using either polyspecific reagent or the anti-IgG antibodies. This study proves that it is possible to perform screening of irregular antibodies on uncoagulated blood samples. This possibility allows automation as blood typing and screening of irregular antibodies can be carried out with the same sample.

[Fetal cells in the maternal blood: a step towards non-invasive prenatal diagnosis? Review of the literature]. / Cellules foetales dans le sang maternel: vers un diagnostic anténatal non invasif? Revue de la littérature.

Prenatal diagnosis of genetic abnormalities requires nucleated fetal cells which are currently obtained by invasive techniques such as amniocentesis, chorionic villus sampling and percutaneous umbilical blood sampling. Each of these entails a risk to the foetus and sometimes to the mother. Nucleated fetal cells have been reported to be present in maternal blood. Recovery of fetal cells from maternal blood would allow a noninvasive prenatal diagnosis. Their rarity (1 fetal cell for 10(6) to 10(8) maternal cells) presents a technical challenge. Due to the small number of fetal cells, sensitive analysis techniques such as PCR and FISH are necessary. Some degree of fetal cells enrichment in the maternal blood sample often precedes the analysis. Different techniques are used for the enrichment: discontinuous density gradient, magnetic activated cell sorting, fluorescence activated cell sorting, micromanipulator.... Several prenatal diagnosis have already been performed from maternal venous blood samples: diagnosis of gender, RhD blood genotype, Duchenne muscular dystrophy and hemoglobinopathy by PCR, diagnosis of gender and chromosome aneuploidy by FISH. Many teams are working on this subject. It is difficult to compare the studies because the techniques of enrichment and analysis vary. We review the different strategies chosen for prenatal diagnosis from maternal blood and discuss the results.