phoenix_1619

Great thanks to you all for the replies i got a good ideas and i'll do some of it today. i called the product specialist in Diamed and he is coming to help me in resolving this flocculation issue !! my pathologist used to work in a hospital that use the gel long time ago but this is her fourth year here in this hospital that use tube method. she said that the positive is showing a clear band . our patient is not black and i'll send her sample for genotyping in her next visit if DAT remain positive. thanks again and see you later

Hello to every body.. Please be patient while reading this long thread... but I really need your help. we have a 4 years old patient with sickle cell anemia and multiple sequestration for seplenectomy her previous history in the referring hospital is that she is A RhD pos ( cells are reacting only 1+ with anti D) by tube method and she has been transfused 12 times by A RhD Negative and she has no phenotype , her previous ab screen was Negative all the times . The last time that they did it was the first of April and she had her transfusion with no problems. She was admitted on the third of May with positive Ab scr ,3 cells were 1+ and AC 1+, DAT was 1+, IgG 1+, C3d Neg .Ab ID has one cell negative which match with the (e) , I could not rule out the C, K,S, Fya, JKa and e after acid elution a panel was run and all the 11 cells were reacting 3+. her phenotype was R0r , i.e: E,C Negative and e, c Positive , this was done by Diamed Id cards using her cells after treating with hypotonic saline to hemolyse the donor cells ( there were actually two cell populations) so our pathologist advice her doctor to wait for one month later to have a clear picture of her phenotype. At this time we started to use the cards for most of our testing. After she came again last week through OPD clinic we phenotype her again , the Rh phenotype was same as before R0r she is Kell , M,N,Jka,Jkb, P1 positive , i couldn't phenotype the Ss , k & Duffys because her DAT was positive. i tried the old fashion gentle heat elution method to elute the Ab while saving the integrity of the cell but it didn't work, the DAT was still positive. her Ab scr, AC, DAT, IgG, Ab Id are all reacting 2+ by the cards , I did the auto adsorption by PEG but only twice not triple ( i have little cells) the auto adsorbed sera was reacting exactly as the little e ( one cell negative when (e) is Negative , one cell is 1+ when (e) and E are both positive "heterozygous" while all other cells are reacting 2+ Sorry for the details and now coming to my Questions: 1- our pathologist said that the reaction in the cards looks like flocculation not true agglutination and is not convinced that it is auto Ab with specificity because the patient is not hemolysing ( we are new to the gel method so could some one explain this flocculation issue to me please?) 2- What is the best way to phenotype the patient cells? If possible kindly suggest a reagent that can help with the name of its manufacturer, so we can order it for future use. 3- What is the best choice of phenotype that I have to give her if transfusion is decided? Thanks in advance to all the helpful ideas:confused: Eman

I appreciate your sharing with us thanks a million Eoin.