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comment_35007

Some times we have a blood donor whose Direct Rh test is Negative but his weak Rh test (Du) positive while DCT/Rh Control also positive.

We try gentle heat elution but usually it is of no benefit.

What is the best technique to determine his Rh.

Do u face this problem?

Thanks

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comment_35009

Sounds like your donor has a positive Direct Antiglobulin test (direct Coombs'). The unit should not be released to a transfusion service. If you really want to know you can try to absorb and elute anti-D from the red cells.

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comment_35012

sure we discard the unit, but what result for Rh for the Donors. Please refer me to the procedure for Absorption and elution for anti D. I try gentle heat elution and rarely I get DCT positive and the problem is solved but it is not always successful.

comment_35015

Hi khalidm3,

Do you have access to chloroquine diphosphate? If you do there is a procedure in AABB Technical Manual for Dissociation of IgG by Chloroquine for Red Cell Antigen Testing of Red Cells with a Positive DAT.

comment_35031

Just generically - rx equal volumes of your donor red cells and anti-D; also run using Rh+ and Rh= cells as controls. (1 mL should suffice). I'd incubate at 37C for 15 minutes, but that is up to you. Wash the cells 6x. Perform standard elution on all and test eluate for anti-D reactivity (also test the eluate from Rh+ and Rh= cells: the Rh+ should show anti-D, the Rh= should show no D reactivity). Also remember to test each last wash as a control for your elution. I cannot point you to a formal procedure for this.

Anybody else?

sure we discard the unit, but what result for Rh for the Donors. Please refer me to the procedure for Absorption and elution for anti D. I try gentle heat elution and rarely I get DCT positive and the problem is solved but it is not always successful.
  • Author
comment_35032

Thanks David, I am used to do adsorption-elution for week A or B, but this the excellent thought from you.

comment_35060
Have you consider a molecular test for identifying/confirming RhD status?

I thought that, but not everyone can get access to this technology.

comment_35061

Thanks Malcolm.

That is true and I thought of it right after sending my message. However, there are a lot of labs around the world that could provide that service. The closest to Riyadh, where I believe khalidm3 is based, that I know is in Kuwait.

comment_35062
Thanks Malcolm.

That is true and I thought of it right after sending my message. However, there are a lot of labs around the world that could provide that service. The closest to Riyadh, where I believe khalidm3 is based, that I know is in Kuwait.

True.

  • Author
comment_35067
I thought that, but not everyone can get access to this technology.

Same here, it is just in our thoughts.

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comment_35091
do you have immucor supplier in your area? They have chloroquin...

Yes, but I am working in a governmental hospital, purchase is not so easy. I will try David procedure, more appealing to me.

comment_35139

You know, everyone is so hot for molecular testing . . . it sounds really neat BUT everytime I investigate it the cost is prohibitive . . . esp for determining the Rh of a donor - seems exhorbitant.

comment_35146

Before I retired, my blood center was charging around $650 which was the amount that hospitals could recoup from insurance payments. If you're talking about investigating patients with warm autoantibodies, the cost of multiple absorptions and cell treatments to remove IgG vs molecular testing could work out about the same.

comment_35171
You know, everyone is so hot for molecular testing . . . it sounds really neat BUT everytime I investigate it the cost is prohibitive . . . esp for determining the Rh of a donor - seems exhorbitant.

Molecular testing can be costly, true, but what is the cost of reliable identifying Rh on a donor such as the one of this case?

comment_35172

I am not going to use the donor so I don't really care what the Rh type is . . . if the DAT is negative next visit, the Rh will be uncomplicated; if the Rh is still problematic next visit, I'm still not going to use the donor . . . kind of pragmatic but I wouldn't bother in this instance.

Molecular testing can be costly, true, but what is the cost of reliable identifying Rh on a donor such as the one of this case?
comment_35173
I am not going to use the donor so I don't really care what the Rh type is . . . if the DAT is negative next visit, the Rh will be uncomplicated; if the Rh is still problematic next visit, I'm still not going to use the donor . . . kind of pragmatic but I wouldn't bother in this instance.

Hi David,

Thanks for your message.

That's a good way of looking at the issue. Certainly, very pragmatic and what most blood banks do.

However, what's the cost of that blood unit you're disposing? And what's the cost of the 2 blood units you'd be disposing if the donor returns? Wouldn't just that justify the investment in a molecular test?

Sorry I'm playing the devil's advocate here, but I'm enjoying the discussion. Again thanks for your views.

Regards

comment_35175

Just to be another devil, what would be the cost if the donor turned out to be D positive, but the unit was transfused into a D negative female of child-bearing potential, she made an anti-D, lost the baby and then sued????????

I am very much in agreement with David on this one.

Take NO chances.

comment_35213
Just generically - rx equal volumes of your donor red cells and anti-D; also run using Rh+ and Rh= cells as controls. (1 mL should suffice). I'd incubate at 37C for 15 minutes, but that is up to you. Wash the cells 6x. Perform standard elution on all and test eluate for anti-D reactivity (also test the eluate from Rh+ and Rh= cells: the Rh+ should show anti-D, the Rh= should show no D reactivity). Also remember to test each last wash as a control for your elution. I cannot point you to a formal procedure for this.

Anybody else?

Assuming the initial problem was a positive direct antiglobulin test due to an autoantibody, most likely a panagglutinin, isn't there a good chance that the eluate will react with all panel cells due to the autoantibody, making it difficult to see the anti-D in there? If there isn't a clearcut specificity for anti-D, one might want to titer the eluate against a Rh+ cell and a Rh- cell in hopes of finding better a higher titer (or not) against the Rh+ cell.

This is, of course, to try to satisfy the insatiable blood banker's curiosity that we all possess. We want to know the answer to the problem! As was stated, you're not going to use the unit. (And I must admit that we do have a computer entry choice for blood type results, used with extreme rarity, reluctance and sadness, "Unable to determine".)

comment_35214
If there isn't a clearcut specificity for anti-D, one might want to titer the eluate against a Rh+ cell and a Rh- cell in hopes of finding better a higher titer (or not) against the Rh+ cell.

The trouble is, an auto-anti-LW, more common than most people think, will give very similar results, and you still will not know if the red cells are D+ or D-.

Sorry to muddy the waters.

comment_35218

Good point, Malcolm. You once called anti-P1 the forgotten antibody, I think anti-LW might qualify as well.

comment_35231

I actually sent Khalid a private message to this effect. It might be medication induced, in which case there is a reasonable chance that it may not be a "pan". But I did warn him anyway . . . JIC.

Assuming the initial problem was a positive direct antiglobulin test due to an autoantibody, most likely a panagglutinin, isn't there a good chance that the eluate will react with all panel cells due to the autoantibody, making it difficult to see the anti-D in there? If there isn't a clearcut specificity for anti-D, one might want to titer the eluate against a Rh+ cell and a Rh- cell in hopes of finding better a higher titer (or not) against the Rh+ cell.

This is, of course, to try to satisfy the insatiable blood banker's curiosity that we all possess. We want to know the answer to the problem! As was stated, you're not going to use the unit. (And I must admit that we do have a computer entry choice for blood type results, used with extreme rarity, reluctance and sadness, "Unable to determine".)

  • Author
comment_35270
Assuming the initial problem was a positive direct antiglobulin test due to an autoantibody, most likely a panagglutinin, isn't there a good chance that the eluate will react with all panel cells due to the autoantibody, making it difficult to see the anti-D in there? If there isn't a clearcut specificity for anti-D, one might want to titer the eluate against a Rh+ cell and a Rh- cell in hopes of finding better a higher titer (or not) against the Rh+ cell.

This is, of course, to try to satisfy the insatiable blood banker's curiosity that we all possess. We want to know the answer to the problem! As was stated, you're not going to use the unit. (And I must admit that we do have a computer entry choice for blood type results, used with extreme rarity, reluctance and sadness, "Unable to determine".)

If panagglutinin, what will be the ICT (ABS)?

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