Bb_in_the_rain

It would appear that Alan is on annual leave outside the UK at the moment, but don't give up.  He has to come back at some time (unless he has won the national lottery)!

When we have these in our central transfusion services, we look into it a bit deeper here in IRL. We repeat tube method using 3 different sources of anti-D. If all 3 sources were negative, report RhD negative. If all 3 sources were >2+, report RhD positive. If 1 of 3 sources were <2+, refer to genomic testing looking for WeakD type 1, type 2 or type 3 (if patient is female childbearing age or <18 year old). Genomic WeakD type 1, 2 or 3 were treat it as RhD Positive. If not genomic WeakD type 1, 2 or 3, treat it as RhD negative. 
If 1 of 3 sources were <2+, all 3 sources give positive reactions and patient does not have anti-D and patient is not female of childbearing age or pediatric (<18 year old), report as RhD positive. (if the patient, later on, makes anti-D change it to RhD negative)
 
Phew.!! I think the decision overall is more based on the patient's age and demographic.

I will touch base with Alan Gray (who is the real expert in this field - you will see, that it was he who wrote the SOPs) and get your questions answered, but, as both he and I are officially retired, it might take a couple of days for us to contact one another.
Thank you for your kind comments.

Works for me. I'll talk to Laurie Munk at AABB and see if there's a good way to get the word out about a sign-up sheet. And make sure I'm not breaking any association rules..
I hadn't heard about Marion Scott retiring!! I was talking to Jill Storry last week and she told me about attending  Geoff's party. It is frightening - everyone I trained with has retired. Of course, a lot of the Canadians could bail at 55 so that's not fair I feel like a dinosaur at AABB. So many new faces!
Will keep you posted as I get information

Just a matter of being in the right place at the right time. I'm getting ready to bail and Malcolm has retired so just think of all the stuff that is going to happen that you'll see and we'll just audit. Get out to the meetings if you can and talk to the speakers. It's an interesting time to be in this field
 

I'm jealous.  You two know all the rock stars.  

I think I am correct in saying that Peter Issitt set up Montgomery Scientific Publication specifically to print the 4th edition (I'm not certain, mind you).
I am not at work until 04/01/16 (UK date format!), but after that I can get in touch with Dave Anstee and find out.
You may be able to get a copy via Amazon, or somewhere like that (but what state it would be in - slightly foxed as a minimum, I would say - I hesitate to think!).

Hello, 
Great discussion. I just set up a comparison study using DTT made with different pH buffer and here is the data (a copy of poster to be posted at 2017 AABB meeting). Hope it help answering some questions here. 
to print DTT poster AABB 2017.pdf

Hello, 
Great discussion. I just set up a comparison study using DTT made with different pH buffer and here is the data (a copy of poster to be posted at 2017 AABB meeting). Hope it help answering some questions here. 
to print DTT poster AABB 2017.pdf

Here is the quick and dirty for the stroma preparation procedure:
We used three units of red cells (R1R1, R2R2, rr; one homozygous for Jka, one for Jkb, one for S, one for s; not more than one unit positive for K). Each unit was treated with 10 cc of 1% ficin (freshly prepared) in a waterbath for 15 minutes. Test after incubation with glycine soya to ensure 2+ reaction. Incubate longer if needed. Wash 3 times in a Cobe 2991 with no red cell override. Add 75 cc of digitonin, mix, and incubate in the processor for 5 to 15 minutes. Do three manual washes with superout set to 300cc. Add 75 cc of digitonin, mix, and incubate in the processor for 5 to 15 minutes. Wash until stroma is white (a minimum of 2 liters saline - watch the waste bag, it can't take this much!) Spin times at least 3 minutes gradually increase the superout as the hemolysis is reduced. Additional digitonin may be added if the stroma is not the desired whiteness. Be sure to wash thoroughly afterwards.
Test the supernatant with red cells to see if all the digitonin was removed. The stroma may require extra washing before it is used if these cells hemolyze.
After wash is completed, add about 200 cc saline.
Aliquot into tubes and freeze at -70C Use within 6 months.
Credit the staff at Florida's Blood Centers Reference Lab for this procedure.

I have recently done an oral presentation about our research project on anti-CD38(Daratumumab) during this recent AABB meeting in Anaheim. I have some abstract references that I have attached here. Hope it helps. 
Also a full article here. http://onlinelibrary.wiley.com/doi/10.1111/trf.13069/full
3 of 8 patients(i have seen to date) has positive DAT and 2 of them were on IVIG. We found that this antibody is non-reactive with In(Lu) cells by tube method. By tube method, DTT treated cells were non reactive, cord cells were weakly reactive to non-reactive.(not enough cord cells were tested yet to find pattern) 
Our flow cytometry data wiith florusecnt labeled anti-CD38 shows that DTT treated cells has very low percentage of CD38 (2%-4%), cord cells has about 21%-24% and positive control varies from (53-82%), In(Lu) cells has about >20%. These data were presented orally during AABB meeting but not yet been published. 
 
SKMBT_75115100213120.pdf
NEJM Aug 2015 editorial on Dara.pdf
cord rule out.pdf
abstract-ARC and NYBC.pdf
abstract-ARC and NYBC.pdf

I have recently done an oral presentation about our research project on anti-CD38(Daratumumab) during this recent AABB meeting in Anaheim. I have some abstract references that I have attached here. Hope it helps. 
Also a full article here. http://onlinelibrary.wiley.com/doi/10.1111/trf.13069/full
3 of 8 patients(i have seen to date) has positive DAT and 2 of them were on IVIG. We found that this antibody is non-reactive with In(Lu) cells by tube method. By tube method, DTT treated cells were non reactive, cord cells were weakly reactive to non-reactive.(not enough cord cells were tested yet to find pattern) 
Our flow cytometry data wiith florusecnt labeled anti-CD38 shows that DTT treated cells has very low percentage of CD38 (2%-4%), cord cells has about 21%-24% and positive control varies from (53-82%), In(Lu) cells has about >20%. These data were presented orally during AABB meeting but not yet been published. 
 
SKMBT_75115100213120.pdf
NEJM Aug 2015 editorial on Dara.pdf
cord rule out.pdf
abstract-ARC and NYBC.pdf
abstract-ARC and NYBC.pdf

I have recently done an oral presentation about our research project on anti-CD38(Daratumumab) during this recent AABB meeting in Anaheim. I have some abstract references that I have attached here. Hope it helps. 
Also a full article here. http://onlinelibrary.wiley.com/doi/10.1111/trf.13069/full
3 of 8 patients(i have seen to date) has positive DAT and 2 of them were on IVIG. We found that this antibody is non-reactive with In(Lu) cells by tube method. By tube method, DTT treated cells were non reactive, cord cells were weakly reactive to non-reactive.(not enough cord cells were tested yet to find pattern) 
Our flow cytometry data wiith florusecnt labeled anti-CD38 shows that DTT treated cells has very low percentage of CD38 (2%-4%), cord cells has about 21%-24% and positive control varies from (53-82%), In(Lu) cells has about >20%. These data were presented orally during AABB meeting but not yet been published. 
 
SKMBT_75115100213120.pdf
NEJM Aug 2015 editorial on Dara.pdf
cord rule out.pdf
abstract-ARC and NYBC.pdf
abstract-ARC and NYBC.pdf

I am guessing red cells doesnt get destroyed because CD38 expression on red cells are very low? 

I have recently done an oral presentation about our research project on anti-CD38(Daratumumab) during this recent AABB meeting in Anaheim. I have some abstract references that I have attached here. Hope it helps. 
Also a full article here. http://onlinelibrary.wiley.com/doi/10.1111/trf.13069/full
3 of 8 patients(i have seen to date) has positive DAT and 2 of them were on IVIG. We found that this antibody is non-reactive with In(Lu) cells by tube method. By tube method, DTT treated cells were non reactive, cord cells were weakly reactive to non-reactive.(not enough cord cells were tested yet to find pattern) 
Our flow cytometry data wiith florusecnt labeled anti-CD38 shows that DTT treated cells has very low percentage of CD38 (2%-4%), cord cells has about 21%-24% and positive control varies from (53-82%), In(Lu) cells has about >20%. These data were presented orally during AABB meeting but not yet been published. 
 
SKMBT_75115100213120.pdf
NEJM Aug 2015 editorial on Dara.pdf
cord rule out.pdf
abstract-ARC and NYBC.pdf
abstract-ARC and NYBC.pdf

I think it depends on the method used. ECHO solid phase has shown to have weak to negative reactions with screen cells and panel cells from what I have seen so far. But non-the-less they were positive screens (weakly reactive but not all cells were reactive) that we had to work up an antibody identification. By tube method. it is reactive with PeG, LISS and saline IAT. 

I have recently done an oral presentation about our research project on anti-CD38(Daratumumab) during this recent AABB meeting in Anaheim. I have some abstract references that I have attached here. Hope it helps. 
Also a full article here. http://onlinelibrary.wiley.com/doi/10.1111/trf.13069/full
3 of 8 patients(i have seen to date) has positive DAT and 2 of them were on IVIG. We found that this antibody is non-reactive with In(Lu) cells by tube method. By tube method, DTT treated cells were non reactive, cord cells were weakly reactive to non-reactive.(not enough cord cells were tested yet to find pattern) 
Our flow cytometry data wiith florusecnt labeled anti-CD38 shows that DTT treated cells has very low percentage of CD38 (2%-4%), cord cells has about 21%-24% and positive control varies from (53-82%), In(Lu) cells has about >20%. These data were presented orally during AABB meeting but not yet been published. 
 
SKMBT_75115100213120.pdf
NEJM Aug 2015 editorial on Dara.pdf
cord rule out.pdf
abstract-ARC and NYBC.pdf
abstract-ARC and NYBC.pdf

I think it depends on the method used. ECHO solid phase has shown to have weak to negative reactions with screen cells and panel cells from what I have seen so far. But non-the-less they were positive screens (weakly reactive but not all cells were reactive) that we had to work up an antibody identification. By tube method. it is reactive with PeG, LISS and saline IAT. 

I am guessing red cells doesnt get destroyed because CD38 expression on red cells are very low? 

This antibody will not be adsorbed out since the dose is very high relative to CD38 expression on RBC. 

I have recently done an oral presentation about our research project on anti-CD38(Daratumumab) during this recent AABB meeting in Anaheim. I have some abstract references that I have attached here. Hope it helps. 
Also a full article here. http://onlinelibrary.wiley.com/doi/10.1111/trf.13069/full
3 of 8 patients(i have seen to date) has positive DAT and 2 of them were on IVIG. We found that this antibody is non-reactive with In(Lu) cells by tube method. By tube method, DTT treated cells were non reactive, cord cells were weakly reactive to non-reactive.(not enough cord cells were tested yet to find pattern) 
Our flow cytometry data wiith florusecnt labeled anti-CD38 shows that DTT treated cells has very low percentage of CD38 (2%-4%), cord cells has about 21%-24% and positive control varies from (53-82%), In(Lu) cells has about >20%. These data were presented orally during AABB meeting but not yet been published. 
 
SKMBT_75115100213120.pdf
NEJM Aug 2015 editorial on Dara.pdf
cord rule out.pdf
abstract-ARC and NYBC.pdf
abstract-ARC and NYBC.pdf

This antibody will not be adsorbed out since the dose is very high relative to CD38 expression on RBC. 

I think it depends on the method used. ECHO solid phase has shown to have weak to negative reactions with screen cells and panel cells from what I have seen so far. But non-the-less they were positive screens (weakly reactive but not all cells were reactive) that we had to work up an antibody identification. By tube method. it is reactive with PeG, LISS and saline IAT. 

I have recently done an oral presentation about our research project on anti-CD38(Daratumumab) during this recent AABB meeting in Anaheim. I have some abstract references that I have attached here. Hope it helps. 
Also a full article here. http://onlinelibrary.wiley.com/doi/10.1111/trf.13069/full
3 of 8 patients(i have seen to date) has positive DAT and 2 of them were on IVIG. We found that this antibody is non-reactive with In(Lu) cells by tube method. By tube method, DTT treated cells were non reactive, cord cells were weakly reactive to non-reactive.(not enough cord cells were tested yet to find pattern) 
Our flow cytometry data wiith florusecnt labeled anti-CD38 shows that DTT treated cells has very low percentage of CD38 (2%-4%), cord cells has about 21%-24% and positive control varies from (53-82%), In(Lu) cells has about >20%. These data were presented orally during AABB meeting but not yet been published. 
 
SKMBT_75115100213120.pdf
NEJM Aug 2015 editorial on Dara.pdf
cord rule out.pdf
abstract-ARC and NYBC.pdf
abstract-ARC and NYBC.pdf

I just passed the test yesterday! I am hugely relieved. I studied the Gulf Coast last chance review, the AABB notes on sbb and the physicians handbook. All are great sources and would recommend to learn them all!