QC AHG, C#/ Part II

[Brenda K Hutson]

I submitted a question yesterday regarding what people are using to QC Anti-C3b, -3d. As I was looking at all of the Manufacturer's Inserts for Coombs Reagents and Check Cells, I came across something else that I have never done anywhere I have worked. It is in relation to QC of Complement Control Cells, but involves Polyspecific AHG.

Along the lines of adding Check Cells to QC, I would also apply this to Anti-IgG. But of interest was something stated in the insert for Complement Control Cells. It stated: Add 1 drop of Immucor Complement Control Cells to a tube containing 1 or 2 drops of Anti-Human Globulin (containing anti-C3; so Polyspecific), for quality control of the reagent (the reagent being Complement Control Cells).

As I think about that, it does make sense. I mean we add Coombs Control Cells to check the IgG aspect of Polyspecific Coombs, but what about the Complement aspect?

So, question is: does anyone out there perform this as QC for either Polyspecific AHG (in addition to Coombs Control Cells), and/or Complement Control Cells?

Thanks,

Brenda Hutson, CLS(ASCP)SBB

[Guest MSchanen]

We started performing this testing on the polyspecific AHG reagent after a CAP inspection pointed out exactly what you just stated.

[David Saikin]

This question came up in the past . . . if the IgG sensitized cells worked with poly AHG, it was felt that that sufficed. I will quote the note under TRM.40210 (which discusses the use of complement check cells) . . . "If polyspecific antiglobulin is used, refer to question TRM.40200." TRM.40200 discusses the use of IgG sensitized red cells. Sometimes inspectors are ignorant. One must remember that CAP uses peer review and some do NOT always have strong backgrounds in BB. One must feel confident in the processes in place to support your operation when faced with the lack of technical understanding or even the poor interpretation of CAP standards. Otherwise you will hem and haw which ever way the wind blows (or the inspector dictates).

[BSIPHERD]

We started performing this testing on the polyspecific AHG reagent after a CAP inspection pointed out exactly what you just stated.

When we realized (about a year ago) that some CAP inspectors were doing this, we realized that for tube polyspecific DAT we were going to have to do it twice if negative - one to add Complement Coated Cells to and one to add IgG coated cells to. So we decided to discontinue polyspecific DAT. When we have a DAT ordered now, we perform it with Anti-IgG and with Anti-C3.

[bb1882]

Be sure to check the insert for the polyspecific antisera that you are using. Immucor's Anti-IgG,C3d Polyspecific Murine Monoclonal reagent specifically states that "confirmation that satisfactory levels of both anti-IgG and and anti-complement are present in this reagent should be included as part of the daily quality control rountine."

[Brenda K Hutson]

This question came up in the past . . . if the IgG sensitized cells worked with poly AHG, it was felt that that sufficed. I will quote the note under TRM.40210 (which discusses the use of complement check cells) . . . "If polyspecific antiglobulin is used, refer to question TRM.40200." TRM.40200 discusses the use of IgG sensitized red cells. Sometimes inspectors are ignorant. One must remember that CAP uses peer review and some do NOT always have strong backgrounds in BB. One must feel confident in the processes in place to support your operation when faced with the lack of technical understanding or even the poor interpretation of CAP standards. Otherwise you will hem and haw which ever way the wind blows (or the inspector dictates).

I do agree with your comments regarding CAP Inspections and Inspectors. That being said, in spite of the wording of the CAP Checklist, I am still left with the statement on the Manufacturer's Insert for Complement Control Cells, stating to: Add 1 drop of Immucor Complement Control Cells to a tube containing 1 or 2 drops of Anti-Human Globulin (containing anti-C3; so Polyspecific), for quality control of the reagent (the reagent being Complement Control Cells).

Brenda Hutson, CLS(ASCP)SBB

[Brenda K Hutson]

Be sure to check the insert for the polyspecific antisera that you are using. Immucor's Anti-IgG,C3d Polyspecific Murine Monoclonal reagent specifically states that "confirmation that satisfactory levels of both anti-IgG and and anti-complement are present in this reagent should be included as part of the daily quality control rountine."

I am looking at that Insert but do not see the quote you are referring to. My version of the Insert is dated March 2005; is this the one you are referring to also? Under what header is the comment?

Thanks,

Brenda Hutson, CLS(ASCP)SBB

[David Saikin]

That's what makes America great . . . you can do what you think you need to do. I never stop anyone from doing more than they have to. PS - I no longer use polyAHG so for me, it's a moot point.

[Brenda K Hutson]

That's what makes America great . . . you can do what you think you need to do. I never stop anyone from doing more than they have to. PS - I no longer use polyAHG so for me, it's a moot point.

Hmm...you are the 2nd person to say you stopped using Poly AHG. So, don't care about C' anymore?? I would love to hear more about where you guys are coming from on this.

Also, believe me, I don't want to just do what I "think" I am supposed to do. I am really trying to understand what is required and not do more than necessary. I have never noticed that statement before on that Insert (not sure if it is new, or I just never saw it). So just out of curiousity, if you were still using Poly, and if you saw the statement I referred to on the Insert for Complement Control Cells, would you test them with the Poly for QC, or not?? Just trying to get a feel for what direction to go.

Thanks for you input,

Brenda Hutson, CLS(ASCP)SBB

[Brenda K Hutson]

That's what makes America great . . . you can do what you think you need to do. I never stop anyone from doing more than they have to. PS - I no longer use polyAHG so for me, it's a moot point.

By the way, I not only agree with your comments regarding CAP Inspections and Inspectors, but I DO also agree with your interpretation of the CAP checklist. Again, I am just "stuck" on what to do with the statement on the Manufacturer's Insert.

Thanks again,

Brenda Hutson, CLS(ASCP)SBB

[bb1882]

I am looking at that Insert but do not see the quote you are referring to. My version of the Insert is dated March 2005; is this the one you are referring to also? Under what header is the comment?

Thanks,

Brenda Hutson, CLS(ASCP)SBB

The one we have was last revised 10/2007. If I remember correclty that is when this was put into the insert. It is under the Quality Control heading of the insert.

[Brenda K Hutson]

Be sure to check the insert for the polyspecific antisera that you are using. Immucor's Anti-IgG,C3d Polyspecific Murine Monoclonal reagent specifically states that "confirmation that satisfactory levels of both anti-IgG and and anti-complement are present in this reagent should be included as part of the daily quality control rountine."

Ok, so it helps to pay attention to the "Manufacturer" stated! My insert is from Ortho, not Immucor. But on the positive side, it means I don't have an outdated insert in my binder (which is what I thought).

I will try to look up your insert just to compare to Ortho.

Thanks,

Brenda Hutson, CLS(ASCP)SBB

[David Saikin]

By the way, I not only agree with your comments regarding CAP Inspections and Inspectors, but I DO also agree with your interpretation of the CAP checklist. Again, I am just "stuck" on what to do with the statement on the Manufacturer's Insert.

Thanks again,

Brenda Hutson, CLS(ASCP)SBB

Brenda - I use anti-IgG gel cards and I test for anti-Complement activity using anti-C3b,-C3d with the buffered gel card.

[BSIPHERD]

Brenda - I use anti-IgG gel cards and I test for anti-Complement activity using anti-C3b,-C3d with the buffered gel card.

Is there any way you can share your procedure using the buffered gel cards? I'd really like to go that route. What did you do to validate the procedure and are there references available?

Thanks for any help.

[mcgouc]

I tried this QC for a while, doing the polyspecific in duplicate, and then decided to drop the polyspecific AHG testing. When we got a positive test we had to wash more cells and repeat with the monospecific reagents anyway.

[JerryB]

Am aware of both the checklist and mgf. package insert. For the past few years or so, when we use polyspecific AHG, we routinely set up a tube to test the reactivity of the compliment component in the polyspecific AHG also. The mfg. suggests it and CAP requires it.

[David Saikin]

Sooooo, do you run your complement check cells once per day, with your reagent qc, OR do you set up everything in duplicate so that you can check with both IgG and complement sensitized red cells?

[Mary**]

I did the same thing, especially when I had to purchase a 10 pack of poly and never used it up.

[David Saikin]

I do not believe CAP requires it . . . the standard for complement antiglobulin reagents tells you that if you use poly AHG, refer to the standard on IgG sensitized red cells. It does not say that you have to check your poly AHG with C sensitized cells. As I rather acerbically stated above, if that was the case, you would have to set all your tube testing up in duplicate and confirm with both types of check cells . . . and CAP is not saying that. The standard TRM040210 is asking about anti-C3 reagents. The note is what is obscuring this, because it is talking about reagent with anti-C3 reactivity (which does include poly). Anyway, I am in contact with CAP and their immediate feeling is that with poly, only the IgG sensitized cells are required BUT they will get back to me with a definitive answer - either today or early next week.

[GilTphoto]

Most of the reason for discontinuing the use of Polyspecific has been the high cost and Immucor's policy of forcing you to buy 10 bottles when you would only use 1-2 bottles a year.

When they order a DAT on an adult, you test and report both IgG and C3. Anti-C3 can still be bought 1 bottle at a time.

Anti-IgG and check cells are QC'd daily.

Anti-C3 with Complement check cells are only QC'd as needed.

Even if you use Polyspecific, you would only need to QC the anti-C3 and Comp check cells when you have a patient with a pos DAT

[Brenda K Hutson]

I like the idea of just doing IgG and C' testing; not doing Poly at all. We don't do a lot of DATs anyway (more on Cord specimens, and we just use IgG for that; positive Autocontrols on Antibody work-ups; not a lot of individual DAT orders on adults). In addition, for some reason, the Table in our computer system is set up to require IgG testing with the Poly (but C3 was made separate; not sure why the builders of the system did that, unless it was a limitation of that system). So, while we may be using more C3 reagent, we would save on the Poly and come out the same with IgG.

Something to think about...

Thanks

Brenda

[JerryB]

Maybe I'm missing the mark regarding this question but, when we perform a DAT, using polyspecific AHG, we routinely check the reactivity of the compliment component of the polyspecific AHG EACH DAY OF USE. This would be the only time we would need to. This has been our policy since 2007. This is only our interpretation of the checklist item.

[David Saikin]

The DEFINITIVE answer from CAP central is that if you use a reagent with anti-complement activity, negative reactions must be validated with the appropriate check cells. I guess that means if you use poly AHG that you have to validate all your negative reactions in duplicate (i.e., with IgG and C sensitized check cells).

JerryB, do you validate your negative tube reactions with both check cells? Do you verify your anti-IgG reactivity ony EACH DAY OF USE, or with each negative tube? You didn't say if you used tube or gel or capture . . . I can see if you use gel/capture the each day of use, but if you use tubes, you would have to check all your negative results in duplicate.

WOW!

[David Saikin]

I hate to beat a dead horse, but I could not live with the answer I got on Tuesday from CAP on this so I called them AGAIN today . . . the new DEFINITIVE answer to this is that anti-IgG and poly AHG reagents have to have negative reactions validated using IgG sensitized red cells; anti-C3d, or anti-C3b,-C3d reagent must be validated using complement sensitized red cells. This is a lot more reasonable and more in line with accepted practice when performing antiglobulin testing in tubes.

[Mary**]

Call them a third time!