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Posts posted by Lingkwyz
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28 minutes ago, R1R2 said:
Sometimes strong, direct agglutinins can cause that mix field appearance in gel like anti M, colds, and rouleaux. Also, since RHIG is not a "real antibody" it may not react the same as a true antibody. Ficin in gel can give weird reactions too, IMO.
The reagent cells were papainized sir. What weird reactions did you experience with ficin-treated cells sir?
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Moving on:
What would be the expanation why Neutral Cards used in Ab ID for enzyme-treated cells does not contain any antihuman Globulin?
-As I understand, antibody screening and identification revolves around IAT..
Pls enlighten.. ^____^
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What would your approach be or your "gameplan" on dealing with a possible Kidd antibody?
My experience recently:
27 YO Female
B Pos
DAT: Neg AC: Neg
Rh Phenotype: R2r K=
Kidd Phenotype: Jk (a+b=)
Solid Phase:
Screen Pos (3 pos grade Jkb + cell)
Antibody Identification: Pos to multiple cells (pointing to Jk b exhibiting dosage phenomenon on some hetero Jkb cells)
Gel Card Method:
Screen and Antibody Panel Negative to both regular and enzyme-treated Cells. Autocontrol Negative.
Tube Method:
Negative to Screen 3 (Panocell 3).
-Sample QNS for further testing.
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What would your approach be or your "gameplan" on dealing with a possible Kidd antibody?
My experience recently:
27 YO Female
B Pos
DAT: Neg AC: Neg
Rh Phenotype: R2r K=
Kidd Phenotype: Jk (a+b=)
Solid Phase:
Screen Pos (3 pos grade Jkb + cell)
Antibody Identification: Pos to multiple cells (pointing to Jk b exhibiting dosage phenomenon on some hetero Jkb cells)
Gel Card Method:
Screen and Antibody Panel Negative to both regular and enzyme-treated Cells. Autocontrol Negative.
Tube Method:
Negative to Screen 3 (Panocell 3).
-Sample QNS for further testing.
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58 minutes ago, David Saikin said:
were all your gel results mf? sometimes poor cell addition can lead to that in the tubules when the cells fall down out of the reaction area.
Was the mf only in the enzymed cells? could indicate a cold ab also.
Well, that would be a long shot sir since all roads point out to Passive anti-D (The history of RhIg, childbearing age female, Rh Neg etc.). Also, the cells reacted to D+ reagent red cells only.
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14 hours ago, R1R2 said:
Me just being curious... but was wondering why all the additional testing after initial solid phase results?
Just ran it for curiousity.. Anyway, gel was our mainstream method before we had solid phase. We got the solid phase around a year ago.
15 hours ago, Malcolm Needs said:There is NO such thing as a "classic" picture of a "Passive Anti-D". Whomsoever told you this is a danger.
I know that our (UK) Guidelines do not apply in Saudi Arabia, but I would urge you to have a look at the BCSH Guidelines on antibody testing in pregnancy and the RCOG Green Top Guideline Number 65, on the same subject, by putting these into your search engine, and they both will confirm what I say (I KNOW the BCSH one will, because I was one of the authors!!!!).
Thanks for the clarification Malcolm.. Will do read on that reference.. Now I can rest in peace.. ^^
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Hi guys.. It's me again.
Would like to start this topic for the multitude of questions popping through my mind as start my life in Bloodbanking. I know a lot of you guys (ehem! you know who you guys are..) are pretty much on the high level bloodbanking, whether skills or knowledge. So, can you spare this chap some of them knowldedge?? ^^
Anyway, lemme start;
I recently had one patient 21 yo female, O Neg, Absc Positive with solid phase. Ab ID in solid phase gave a reaction pointing to Anti-D with reactions as low as 1+, max 2+.
Confirmed the testing through gel card which gave me regular results: weak pos, enzyme: 4+ "mixed-field".
As I was the neophyte, I have been told that this is a "classic" picture of a "Passive Anti-D".
So my question is: Why the mixed field? Why didn't the "passive Anti-D" react to all the cells of the D+ reagent red cells?
Answers are highly appreciated.
Thanks
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On 7/14/2016 at 6:49 PM, macarton said:
We have a collection of different colored highlighters that we use on the panel sheet, different color to mark each suspected antibody and go from there.
Thanks for the input macarton.
I actually am looking for a range or values from the p value that could create a possibility of an antibody above 0.05
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6 hours ago, WisKnow said:
I am not sure if there are 0.2M DTT treated cells readily available commercially if that is what you meant. We prepare our 0.2M DTT and use it to treat the positive cells we selected for antibody workup. It's a piece of work because you have to set up 0.2M DTT quality control by picking a heterozygous E and a K+ cell, antigen type them before and after treatment with 0.2M DTT. Kell group is destroyed by it. The antibody identification is performed using the treated cells and after everything has been ruled out, we still recommend K negative unit.
Thanks for the info WisKnow. Still hoping we get us one of those.
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I wish I could get me one of those 0.2M DTT-treated cells. Our institution is currently serving one patient under DARA. fingers crossed.
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On 6/21/2016 at 10:14 AM, Lingkwyz said:
Hi DCE/ce!
Thanks for the info.
We also do proactively give antigen negative units to possible antibodies, proven or not. Nevertheless, full crossmatch is required for all with an antibody in question.
But my question is, for curiosity purposes, that if the p vaule of 0.05 or lower is a marker of a probabillity of the prescence of an antibody, what would be the maximum p value to consider it as a possibility and thus add it to your detections.
Hope one of our masters sees my dilemma..
Paolo
anyone?
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21 hours ago, David Saikin said:
I also think that one must understand the vagaries of the different test systems. Ortho's gel, for example, has a notoriety of not being the best for detecting the Kidd abs (though I have not encountered this it seems it is fairly well documented, just ask an Immucor rep). Also, you mention that some Jka cells reacted and some did not - you did not mention whether the ones that did not react expressed Jka as homozygous or heterozygous, ditto for the ones that did react. You answered my question about the panel (in solid phase vs gel). I think that is all I have to say except I really dislike the Kidds.
I think, in a routine lab as we are in, these antibodies are a hit-or-miss.. Not having PEG, DTT, ZZAP, adsorption techniques etc. keeps us in the dark on situations like these. I kinda suggested these to our greater management but they are totally apathetic about it.
Yup, I did miss to inform that the screening cells of the solid phase in the pre-transfusion testing pointed out a pattern of Anti-Jka. But on the Ready-ID panel got one cell positive which was homozygous to Jka. Nevertheless, there were approximately more than 3 more Jka pos cells which were negative. Also, on the gel card testing of the same sample, all results were negative to screening and panel 11 which i would say are filled with Jka pos homozygous cells.
Thanks David.
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14 hours ago, Mabel Adams said:
We have a couple of policies that might have helped (although if the tech was convinced that the reactivity in solid phase was a false positive, he might still have missed this--our expectations are powerful). We require (well, there are some exceptions) that 2 double dose (homozygous donor) Jka positive cells not react in gel before we can rule it out. I have seen too many anti-Jka antibodies that react with one cell but not another in gel. We pretty much never rule it out with just "positive" cells. I know I look particularly hard at whether something "sort of" fits a Kidd antibody because of their tendency to be weak to undetectable. If all of the positive reactions are on Kidd double-dose cells, I will be doing some extra work to rule it out. Antigen type the patient, as Malcolm says.
We train and it is in our procedure that the gel card must be "QCd" after running which includes looking for proper centrifugation of the card and whether the cell and plasma volumes are correct. I can't promise that no one ever gets lazy and skips this but then they are not following the procedure.
Some other options would be that gel cards run to follow up on questionable solid phase results have to be incubated for 30 minutes instead of 15 (we have validated up to 40 minutes which is about the maximum allowed I believe in MTS gel). This will sometimes bring out a weak antibody or make it react with more cells. It does not change the non-specifics much usually.
Hi Mabel!
Clearly in this case, the Anti-Jka was missed. Enhancement techniques (e.g increase in incubation time, increase in plasma-cell ratio) isn't a popular idea from where I'm at. Sadly, was on the patient's expense.
I would rather educate myself and ask from you guys on how you would have handled the case. Although this was not my case, but then again, information is everything.
"QC" as you've pointed out might be the visual inspection of gel card materials as you do your test. But, my challenge is that, as a reviewer, how sure are you that none of the components of testing, especially the patient's sample is not omitted? I would rather ask for a proactive approach so situations like these gets controlled/prevented.
Next, I would agree on your input regarding being skeptical on "treacherous" antibodies. For me, (for all its worth actually-hehe) caution must be taken in dumping a positive reaction into the "Nonspecific Reaction" ditch.
Thanks for the input Mabel.
-Paolo
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42 minutes ago, Malcolm Needs said:
Yes, I can see from where you are coming. It can be so difficult on occasions (which is why I have always maintained that we should be paid our worth)!!!!!!!!!!!!!
I could not agree more Malcolm.. True that!!!
As I see, and I hope everyone agrees, working on a "routine" blood bank must have its own system "traps". Cruising through a day of "regular" patient samples sometimes gets you to overlook really tricky ones. In our institution, I would say that we are benefiting on the "Non-specific Reaction" dump. Anything unequivocally an antibody would fall to that dump. Sadly, the situation above fell into that.
As per the fee, nah! I always tell my self that I will be here for the science. Sadly again, my wallet don't feel the same!
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10 minutes ago, Malcolm Needs said:
I would say, particularly with antibodies within the Kidd Blood Group System, which are notorious for becoming labile both in vitro and in vivo, but causing severe delayed haemolytic transfusion reactions due to anamnestic responses, that, if there is a strong index of suspicion, the specificity should be honoured, whether it can be proved or not. The only time that I would not do this is if the patient is proved to be, for example, Jk(a+), and the antibody looks like anti-Jka, or Jk(b+), and the antibody looks like an anti-Jkb. I think it is better to give Jk(a-) or Jk(b-) blood unnecessarily than to find out afterwards, when the patient has a reaction.
This, however, is a personal view, rather than a hard and fast "rule".
Hi Malcolm Needs!
I do agree that prophylactically we must give antigen negative units to suspected antibodies, proven or not. But to this situation, the results gave negative reactions to two Jka positive cells in Solid Phase panel and no reaction to Gel Card both regular and enzyme. Also, this patient have had multiple testings in our lab, all of which were interpreted as "Non-specific" reactions.
I also had to point out that as a person seeing the result, what would be my indicator for the validity of the Pre Transfusion Gel Card results (which were all negative) which was also taken into consideration, thus the "Non-specific" reaction interpretation.
Thanks for the enlightenments guys and hope to learn more from you guys..
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Hi Rapundaa!
Thanks for the input and my bad. See, the antibody screening and the ID was done in Solid Phase. The initial identification had only 2 cells positive (2+), suggestive of anti-Jka but was eventually ruled out because of the nonreactivity of the other Jka pos cells. So, as it seems, the MTOD proceeded to another method which is the Gel Card Method which got all results negative.
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Hi guys,
I haven't been exposed as a blood banker for quite some time, so please bear with me. Recently, our institution had a case of transfusion reaction (lower back pain) with one unit of PRBCs. The testing details are as follows:
History: Non-specific reaction (Multiple encounters)
Pre Transfusion Sample
Solid Phase : 2 of 3 cels 2+ (Positive
Gel Card (Regular) : Negative
Gel Card (Enzyme) : Negative
Dx: Non-Specific Reaction, all siginificant antibodies excluded
Transfusion Reaction Investigation Sample
Solid Phase : 2 of 3 cels 2+ (suggestive with Anti-Jka)
Gel Card (Regular) : Positive (2+) Minimum grade (Clear-cut Anti-Jka)
Gel Card (Enzyme) : Positive (4+) (Clear-cut Anti-Jka)
DAT (Gel Card) : Positive
Elution Testing: Anti-Jka Detected
Dx: Anti-Jka
Analyzing the encounter, me and a colleague, through curiosity re-ran the Pre Transfusion Sample:
Pre Transfusion Sample: (Curiosity Run)
Solid Phase : 2 of 3 cels 2+ (suggestive with Anti-Jka)
Gel Card (Regular) : Positive ; Wp Minimum grade (suggestive with Anti-Jka)
Gel Card (Enzyme) : Positive ; 1+ Minimum grade (suggestive with Anti-Jka)
Taking all details, I could presume that the Gel Card results (both regular and enzyme, which I believe was ran at the same time) of the 1st Pre Transfusion Sample was the jinx. So confidence to the testing method lies to the daily QC of the Gel Card which was ran first thing of the day. Now, I'm asking, if Solid-phase technology could incorporate a positive control and negative control to each strip of testing, why aren't there any on the Gel Card Method? Next, if the Solid-phase technology could also incorporate a "system trap" to ensure the addition of patient's sample to the testing, through it's Capture LISS, why cant the Gel Card do so? I believe that seeing an all-negative resulted panel would be enough for some people for exclusion. Lastly, if reagents aren't available for the Gel card method to ensure the addition of plasma, should our institution enforce a backup procedure that could ensure the reviewer that none of the essential components of the testing (e.g. Patient's plasma) is not missed?
Hoping for the masters..
Thanks !
lingkywz
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Hi DCE/ce!
Thanks for the info.
We also do proactively give antigen negative units to possible antibodies, proven or not. Nevertheless, full crossmatch is required for all with an antibody in question.
But my question is, for curiosity purposes, that if the p vaule of 0.05 or lower is a marker of a probabillity of the prescence of an antibody, what would be the maximum p value to consider it as a possibility and thus add it to your detections.
Hope one of our masters sees my dilemma..
Paolo
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Hi Smiller!
Unfortunately, I have checked everywhere and to no avail. Need expert advice on this.
Thanks for the heads up anyway.
Paolo
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Hi!
If you could not lower the p value to 0.05 or less, what would be the p value cutoff for a possibility of an antibody? Like, to consider as a prophylactic or proactive approach to give antigen negative units to the patient.
Thanks Guys!
Paolo
The Kidds
in Case Studies
Posted · Edited by Lingkwyz
1. Do you run enzyme treated cells on Poly/Coombs? Our SOP only allows us to run enzyme treated cells on neutral.
-I did try to test the patient's plasma with regular cells on Coombs/Poly cards, they were negative. -(if weak pos, it would be very faint for my eyes.)
2. No history taken since the patient was an "out-patient", sorry.