BACKGROUND Blood donors exhibiting a weak D or DEL phenotypical expression may be mistyped D negative by standard serology hence permitting incompatible transfusion to D negative recipients. Molecular methods may overcome these technical limits. Our aim was to estimate the frequency of RHD alleles among the apparently D negative Polish donor population and to characterize its molecular background. STUDY DESIGN AND METHODS Plasma pools collected from 31,200 consecutive Polish donors typed as D negative were tested by real-time PCR for the presence of RHD specific markers located in the intron 4, exons 7 and 10. RHD positive individuals were characterized by PCR or cDNA sequencing and serology. RESULTS Plasma cross-pool strategy revealed 63 RHD positive donors harboring RHD*01N.03(n=17), RHD*15(n=12), RHD*11(n=7), RHD*DEL8(n=3), RHD*01W.2(n=3), RHD-CE(10)(n=3), RHD*01W.3, RHD*01W.9, RHD*01N.05, RHD*01N.07, RHD*01N.23, RHD(IVS1-29G>C) and two novel alleles: RHD*(767C>G)(n=3), RHD*(1029C>A). Among 47 cases available for serology, 27 were shown to express the D antigen CONCLUSION 1/ Plasma cross-pool strategy is a reliable and cost-effective tool for RHD screening. 2/ 0.2% of D negative Polish donors carry some fragments of the RHD gene; all of them were C or E positive. 3/ Almost 60% of the detected RHD alleles may be potentially immunogenic when transfused to a D negative recipient.

In the four tests, correlation between expected and obtained results was appropriate (CCC ̴1). Each test had some advantage and limitation in any clinical situation. Therefore, it is best to have opportunity to perform two or three assays in the laboratory.

Feto-maternal haemorrhage assessment in a woman with a large population of red blood cells containing fetal haemoglobinOcena przecieku płodowo-matczynego u kobiety z dużą populacją krwinek czerwonych zawierających hemoglobinę płodową , haptoglobin, iron, transferrin, ferritin, hepcidin, sTR, HbF, HbA2 were measured. Genes coding the `-and a-globin were sequenced. Materials and Methods: Patient' s samples and artificial mixtures were tested by microscopic Kleihaur-Betke (KB) and flow cytometric (FC) tests with anti-HbF + anti-CA (carbonic anhydrase), and with anti-D. The patient' s blood count with reticulocyte parameters, and concentration of bilirubin Results: It was impossible to distinguish the population of fetal and maternal HbF positive cells using KBT and FC with anti-HbF. Application of anti-CA and anti-D allowed to separate them.Maternal blood haematological and biochemical parameters were normal but HbF was 3.3% of total Hb concentration (normal <1%). There were no mutations in the `-and a-globin genes, but Xmn I polymorphism at -158 position in a-globin gene was detected in the homozygous state. Conclusion

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