marilynm

If you do not have human patient antibodies, and use commercial antisera, I think it is appropriate to use "diluted" antisera labelled "for QC purposes only" and dilute the anti-K and anti-E on day of use and only the amount you need that day. If they are monoclonal, you can probably get a good dilution to give a 2+ reaction on untreated cells and 3-4+ on treated. Interested in hearing other suggestions. Marilynm

The on line Distance Learning SBB program of Janet Vincent's at UTMB in Galveston TX is an excellent well established way to prepare for the SBB. Also, the Last Chance Review held at Gulf Coast in Houston Tx by Clare Wong. Marilynm

From an "old time blood banker", There is no perfect test, as you all are saying. I have found that everyone has their preference of commercial companies and automation technologies and all work, or they would not have been licensed by FDA!! Maybe we need to look at "clinical history" with some of these antibodies, which we are doing in our laboratory. Stay tuned. Marilynm PS I would encourage each of you to look at patient/donor history and do followup studies to see if the antibodies are clinically significant. I am always curious as to why these "unusual" antibodies do what they do. Plus, you can have a nice paper out of it for aabb in New Orleans next year.

Thank you for the postings and it is good to be back in the reference lab and teaching. The younger generation will be the future blood bankers and I have seen some very good prospects out there since I have started back working. Also, this is a very good forum and I enjoy reading the postings and replies. A wide variety of expertise out there. Marilynm

In reply to Immunohematologist, what is critical with your case is "what is the DAT or what does the auto control do?" If the DAT and/or autocontrol are negative, then you have a partial D with alloanti-D. If the DAT and/or autocontrol are positive then you probably have auto antibody with anti-D specificity. I work in a Reference Laboratory, but whenever we have an antibody and no information on the patient was supplied, we call the referring laboratory and request at least transfusion and pregnancy history. In the past, when I was in charge of a reference lab that got samples from screening labs we would call them and have them call the doctor to get the information we needed. It was not easy and not always successful, but we always gave it a try. I do not like reporting out "anti-D in a D positive person" unless I know a history. Marilynm

I assume your lady is Indian? I think we need to look up what category of partial D has been found in your population. Also, whose anti-D are you using? I do not think it is D VI as they usually do not give a 4+ reaction with anti-D. Marilynm

I do not think this is anti-LW. I do not see how strong the reactions were, but if they were strong then Rh D negative cells would react, if it were anti-LW. However, it could be ruled out by testing D negative cord cells, as they would react if it were anti-LW, as D neg cord cells have stronger LW antigen then D neg adult cells. Also, DTT-treating the reactive D positive cells would show if it was anti-LW, as they would be nonreactive DTT-treated. I think this is autoantibody mimicing anti-D specificity. Marilyn m

cudos to Anna Galvania. Great references. I just have a quick question. Have you done the D, C and E typings (along with an Rh control) with IS and extended incubation at RT or 37 (depending on your source of Rh reagents and the package insert) and looked at the typings microscopic for mixed field? You might want to follow the person, because we had a patient years ago who was really an R2r and both the D and E typings were only micro and she appeared to be rr (D-E-) and when her anemia was corrected her and D and E typings were reactive macroscopic. She was the first report of this phenomenon in anemia. MarilynM

Autoantibodies can have almost any blood group specificity. We actually investigated a patient with an autoantibody showing D specificity. The reference is the ARC Immunohematology journal: Immunohematology 1994;10:117-119 by Dzik W, Blank J, Lutz P, Hirose TG, Lomas C and Moulds. M titled "Immune hemolysis following transfusion: a mimicking autoanti-D in a D- patient with alloanti-D." Our patient was D negative but it can occur in D positive patients. There are several references in our paper on autoantibody showing anti-D specificity in D positive patients. Once you have ruled out recent transfusions, medications, etc. then the explanation would be autoantibody. Mmoulds

A very good reference for cord cells and LW antigen, as well as LW in general is the ARC journal Immunohematology, 1992;8:87-93 by JR Storry titled "Review: the LW blood group system". She gives quite a few of the original references to the studies done by Jane Swanson and others. MKMoulds

The reason you test D negative cord cells is because cord cells have stronger LW antigen then adult D negative cells, and I am assuming in her testing that D negative panel cells were nonreactive. It really also depends on the strength of the antibody. If it is very strong (2-3+), then adult D neg cells would react with an anti-LW. If it is weak, then they might not. But D neg cord cells would react with anti-LW. MKMoulds

You can also test D negative cord cells to rule out anti-LW, as they would react if it were anti-LW and not anti-D. Make sure the cord cells have a negative DAT,etc, so they would not give a false positive reaction. This is probably easier then testing DTT treated cells, for a small lab. MKMoulds

One more point, there is a partial D kit out there that can help classify some partial D cells by serology before you go to molecular. MKMoulds PS I saw that you do tube and not gel D typing, so ignore my question on that point. MKMoulds

There are several considerations besides those already suggested, but if the autocontrol is negative then she is probably a partial D, lacks part of the D antigen and has produced alloanti-D. It is important to know what the source of the Anti-D reagent is that was used and what clones are in it. Also, what is the race of the person? Has she had previous pregnancies or has she been transfused. You need to find out what stimulated the anti-D. And, of course, the baby could be at risk if it is D positive. It is probably not a D VI, as most of these partial D category do not give a 4+ reaction with commercial anti-D on IS. I guess it is important also to know if you did the D typing in tube or gel or solid phase. MarilynM

Dear Kerry: thank you so much for your hints. I know that with every technique there are tips and observations that are good to share with others, whether it be tube, solid phase or gel. I have worked with Capture some and do know that it is important to centrifuge the serum/plasma good before putting it on Capture, and I would guess the same would be with gel. We will be doing some comparisons soon between tube, gel and solid phase and I will be interested in what antibodies are identifiable at different time frames. I do know that even though commercial antisera says you can incubate 10-15' (whichever the antibody) but that the time can be extended to 30' in some cases and a cell that has weak expression of the antigen might not react until incubation is done for 30'. Fyx (weakened Fyb expression) might be one example, although some examples of anti-Fyb might not react at all, even after 30'. Sorry for the long reply. I keep forgetting that I said I would try to not be too verbose and I already broke my promise. Marilyn

There are rare cells that are rG (D-C-G+) and some that are r"rG (D-C-E+G+). We have a few in our collection of rare cells that are from Germany and Australia, as well as a few from the US. Most D+ and all C+ cells are G+. There are rare cells that are D+C-G- that are partial D. Marilyn

Dw (Rh23) is NOT associated with weak D type. The w stands for Weil (name of the first proband whose rbcs possessed the low incidence antigen. RBCs that are Dw positive have a partial D antigen of DVa; in Caucasians it is DVaCe and in Blacks it is DVace. I hope that answers your question. Marilyn

I have read all the above comments and examples and wish to say that no one method will detect all antibodies or be perfect every time. I have seen antibodies that react in solid phase that cannot be seen by other methods, the same has been true for gel and also in the tube. How many times have you done something in the tube that was not reproducible, gel too and solid phase? Specific to anti-Jka, although autoanti-Jka is not common, it does occur. You must select a method for antibody detection and identification that is best for your lab, learn to deal with the inconsistencies and try to select blood for the patient according to your SOPs. We use tube routinely in our reference lab but can do gel and soon solid phase and hope to have some data soon on all three methods and compare serological findings with clinical data, which I think is very important. Marilyn PS Words of wisdom from the "old" and "wise"?

Dear Shay: good to hear from you. My work e-mail is marilyn.moulds@lifeshare.org. Write me and let me know where you are working,etc. Marilyn

Hello to Belva, Noel and Christie; good to hear from you. I guess I will find out who is on this BloodBank forum, with the replies and it might be getting a few "hits"! I guess I will have to keep up my reputation and start posting on some of the subjects posted recently! Marilyn

Dear John Staley: I am not sure we want the famous John Judd to be back giving his "words of wisdom"! Are you sure the younger generation would be able to handle it? Marilyn PS I can probably get him to give us "advice" once in a while.

Hello to everyone on the BloodBankTalk forum, especially those who know me. I am back. I retired in June 2006 but just could not stay away from blood banking! I work part time at LifeShare Blood Centers, Shreveport LA as an Immunohematology Specialist in the Reference Laboratory. I will try not to post too often or be too windy. Marilyn Moulds MT(ASCP)SBB, formerly Vice President, Reference and Education, ImmucorGamma.

I posted yesterday, but maybe I did not do it right. what was the source of your anti-Lea? I assume it was monoclonal? If it was, I think in the package insert (Gamma's) it talks about getting typings on red cells of Blacks with monoclonal reagents but not others (human or rabbit). Usually Lea typings are 3-4+, so when you get a 1+ reaction, it is not the usual. Just curious. Also, if you used the reagent by a method not recommended by the manufacturer, then you can get unusual results. The Lewis system was never my favorite and I guess we need a molecular specialist to tell us why the monoclonal anti-Lea reacts in some cases and other sources do not. Marilyn m

Just a comment, since it was said I had not been on a while. Anytime you use different methods and change the formulation of any products, you should not expect to always get the same answers, or even the ones you would like. The old saying "the antibodies do not read the books" is still good today. marilynm

Dear Beverly: you are soo good. Helping blood bankers do simple tests that do not need commercial reagents and/or complicated procedures to make their own reagents. Who says the "old" reliable procedures will not still work today! With blood bankers like you I think I need not worry about the future of blood banking without us "oldies". Marilyn