Does anyone have an eluate blind sample recipe for unknowns that will yield enough sample for an ID?

I've used 9 segments from an Rh neg unit plus 1 segment from an Rh pos unit with about 6-8 drops of anti-D. Let that concoction incubate at 37 C for 15 or 20 minutes. I washed the sample once or twice to remove some of the anti-D, then handed it off to the student. When they followed our SOP, the sample worked just fine. I'm sure you could do something similar w/ different antisera, but I've never tried. I just needed to teach the method, not evaluate for a competency.

I've taken an outpatient sample (so I know it won't be needed again), added 3 drops of anti-s (BioRad - IAT), mixed and put it in the water bath agitating for about an hour.  Works like a charm.  I've also used Coombs control (labeled as a patient sample) - they elute anti-D as expected-much less of an eluate volume but we use gel so there's enough. 

We've been using RhIg.  Always has D, usually has C.  We have not had great luck with commercial antisera or human.  We also have 40 people we need to do, so that can get expensive.

Sometimes I use the Coombs Control. Other times I have antigen type an expired unit for E, C or K and if positive for one of those put a couple of mls in a tube add the corresponding anti-sera, incubate for 30 minutes. I found the another anti-seras don't work so  well.

I have found that the anti-sera that requires coombs phase testing works well to make elutes, but the monoclonal anti-seras that only require RT incubation, don't work as well.  Since that is most of the commercial anti-seras now - it is getting tougher to manufacture this stuff yourself now. 

AMCord's recipe would work well, but always leaves you with a anti-D eluate. 

Cliff - do you use a whole syringe of RhIg?  How do you mix it?  

On 1/29/2019 at 12:04 PM, cswickard said:

Cliff - do you use a whole syringe of RhIg?  How do you mix it?  

I attached our "cheat sheet" for making these competencies.

Eluate.docx