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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.

Despite the generalization of immunoprophylaxis by anti-RH immunoglobulins since 1970 and improved management of at-risk pregnancies, allo-immunization due to the RH1 antigen (formerly known as Rhesus D or Rh D) remains widespread. In fact, anti-RH1 antibodies currently constitute over one-third of the immune antibodies detected after pregnancy. At the same time, allo-immunizations against others antigens than anti-RH1, especially anti-RH4 (anti-c) and anti-KEL1 (anti-Kell) increase. Allo-immunization, its follow-up during pregnancy, and its prevention are therefore still topical, and concern all the pregnant women. Immunohematological tests used in antenatal patients have gone 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. The best method to assess the severity is the determination of the level of fetal hemoglobin after fetal blood sampling but this procedure is not without risk. Since 13 years, it is possible to determine the fetal RHD genotype of using amniocytes and to day directly with maternal plasma. All pregnant women should be blood-typed for ABO-RH-KEL1 and the blood tested for clinically irregular antibodies. The trend in anti-RH levels is more important than the level itself. Manual titration is simple but only provides rough, semiquantitative estimates of anti-RH concentration. Quantitative hemagglutination methods, using auto-analyzers and appropriate anti-RH1 standards, measured in mug/ml, are sensitive, rapid and have acceptable intra-laboratory reproducibility. 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 feto-maternal hemorrhage and now routine antenatal RH prophylaxis at the 28th week of gestation. It has been necessary to synthesize the indications of RH prophylaxis and immunohematological tests to assure an efficient therapeutic prevention.

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.

The French Blood Transfusion Society working group Immunohaematology and the French Blood Transfusion Center of Lille performed two quality control exercises-96-01 and 97-02-in order to evaluate identification performances of alloantibodies associated with an autoantibody recognizing a high frequency antigen. Concerning control 96-01, 83 (75%) of the 110 blood transfusion centers participating at this exercise sent results. The alloantibody screening was correct for 78 of them (94%). Sixty-one (78%) blood transfusion centers correctly identified the specificities (anti-RH1 + anti-FY1). Concerning control 97-02, 82 (94%) of the 87 blood transfusion centers participating in this exercise sent results. The alloantibody screening was correct for 69 (88%) of them. Fifty-three blood transfusion centers (77%) correctly identified the specificities (anti-RH3 + anti-FY1). These exercises allowed us to confirm the main procedures used in routine for national scale testing. The analysis of the results has underlined the importance of these tests for assessing the quality of these examinations, and highlighted the means to be carried out in order to improve them.

This study reports the results of ABO-Rh (D) typing in microplate according to a suggested protocol. 35,532 blood typings were performed by 13 laboratories, compared to usual technics. This work has proved the feasibility in routine of this protocol in order to identify the A, B, D and weak antigens. However the difficulties in detecting some weak variants reveal the interest of standards for immuno-haematology reagents, to apply in the microplate technology.

Peripheral lymphocytes were obtained from an immunized woman against C and Ce antigens. Punction was realized 14 days after childbirth. After hetero-hybridization, a unique cell line continued secreting a monoclonal IgM antibody. Serological characterization of this antibody was determined by direct agglutination tests against 150 native and enzyme treated red blood cells including some rare phenotypes. This antibody was specific of C determinant of the Rh system. It showed strong reactions by saline and enzymatic technics, against C positive cells from Ce positive and Ce negative patients. The validation was performed by a manual direct agglutination test in saline (tube) and by an automated-hemagglutination test (in microplate) against 2,500 patients samples. No discrepancy was observed. This monoclonal IgM anti-C could be used as a potent reagent and since one year, about 30,000 patients and pregnant women have been phenotyped in our laboratory successfully.

During the last four years, 118 blood donors have been immunized to obtain plasma with a high level anti-D in order to prepare anti-D immunoglobulins. The results of the immunizing schedule are very successful, as we have obtained anti-D of titer superior to 256 in 96,36% of the cases (Coombs technic). However, the development of unwanted antibodies outside the Rh system (anti-Jka: 6, anti-Fya: 5) has led us since November 1979 to use phenotyped blood without undesirable red blood cell antigens. No irregular antibody has developed since except for an anti-Yta. The anti-HLA have been observed with a frequency of 36%. The use of frozen/thawed and phenotyped blood without undesirable red blood cell antigens can allow to obtain a high level of anti-D without risk for the donors. Nevertheless, the exceptional immunization to a public antigen persists.

Evolution of idiotypic determinants on lymphocytes membrane and presence of other lymphocytes carrying anti-idiotypic determinants, were studied in Rh negative human volunteer blood donors during immunization towards Rh factor. For this purpose E-Rh Rosettes, direct immunofluorescence, inhibition of E-Rh Rosettes by anti-idiotypic sera as well as EA-Rh Rosettes--induced with Fab'2 fragments from anti-Rh antibodies and lymphocytes from the same subject--were examined and compared to the evolution of circulating antibodies. E-Rh Rosettes preceded or accompanied production of anti-Rh antibodies; their frequency decreased after the fifth month following immunization, meanwhile EA-Rh Rosettes increased in parallel with antibody decrease. The direct immunofluorescence and the inhibition of E-Rh Rosettes, by anti-idiotypic sera, show the presence of idiotypic determinants on lymphocyte membranes; the presence of EA-Rh Rosettes, coinciding with the decrease in antibodies demonstrate the existence of lymphocytes bearing auto-anti-idiotypic determinants.