Gamma Pi Blood Group Substance
(Avian) Gamma Pi Blood Group Substance
|For Assistance in the Identification of Unexpected Blood Group Antibodies)
riyp] RxONLY Q®
No U.S Standard of Potency
Harmful, Preservative: 0.1% Sodium Azide
Do not use if markedly turbid. CAUTION: THE PACKAGING OF THIS PRODUCT
BULBS) MAY CONTAIN DRY NATURAL RUBBER.
(DROPPER
Norcross, GA 30071 USA
r____ _, Immucor Medizinische Diagnostik GmbH
| ec | r e p | Robert-Bosch-Strasse 32
63303 Dreieich, GERMANY 3004-4
INTENDED USE: Gamma P i Blood Group Substance is intended for use in
assistance in the identification of unexpected blood group antibodies by neutralizing
anti-Pi activity in serum or plasma.
SUMMARY OF THE TEST:
The ability of some blood group antibodies to be neutralized by soluble blood group
substance can be utilized to provide confirmation of antibody specificity in certain
cases. An example of a neutralizable antibody is anti-Pi , which may occur without a
red-cell-induced stimulus in Pi-negative persons. The addition of soluble P i Blood
Group Substance to a serum or plasma sample appearing to contain anti-Pi
enables the identity of this specificity to be confirmed, and permits the sample to be
studied for the presence of other antibodies without requiring a panel of red blood
cells selected for the absence of the P i antigen. A potent source of soluble P i
substance has been found in the whites of turtle-dove eggs [1].
PRINCIPLE OF THE TEST: When Gamma P i Blood Group Substance is added in
proper proportions to a serum or plasma sample containing antibodies directed at
the Pi antigen, subsequent testing against P i + red blood cells will yield negative
reactions. By comparing the reactions of substance-treated sample with those of
untreated sample, the identity of anti-Pi may be confirmed. If additional antibodies
are present in the sample, neutralization of the anti-Pi component may enable these
antibodies to be detected and identified. The effect of dilution caused by adding 1
part of P i substance to 10 parts of sample is minimal, but can be measured by
parallel testing of the same sample to which saline has been added in the same 1 to
10 proportions.
REAGENT: Gamma P i Blood Group Substance is a standardized solution of avian
Pi blood group substance in phosphate-buffered saline. Contains 0.1% sodium
azide as a preservative.
PRECAUTIONS:
For in vitro diagnostic use. Store at 1 ° to 10°C when not in use. Do not dilute. Do
not freeze. Do not use if markedly turbid. Effort should be made to minimize
contamination during use of the product. Do not use beyond expiration date.
< f V r h i s reagent contains 0.1% sodium azide. Warning: H302 Harmful
swallowed.
Warning: Sodium azide may react with lead and copper plumbing to form highly
explosive metal azides. If discarded into sinks, flush with a large volume of water to
prevent azide build-up.
Handle and dispose of reagent as potentially infectious.
CAUTION: THE PACKAGING OF THIS PRODUCT (DROPPER BULB) MAY
CONTAIN DRY NATURAL RUBBER.
The format for the expiration date is expressed as CCYY-MM-DD (year-month-day).
SPECIMEN COLLECTION AND PREPARATION: The specimen originally
collected and prepared for antibody detection and identification is suitable for the
test procedure. If there is an insufficient volume of the original spocimon to permit
parallel testing of substance-treated and saline-diluted serum or plasma, a fresh
specimen should be drawn, observing the directions accompanying the Reagent
Red Blood Cells in use.
PROCEDURE:
Materials Provided: Gamma P i Blood Group Substance
Key:
Underline = Addition or significant change ▲ = Deletion of text
(Avian)
|For Assistance in the Identification of Unexpected Blood Group Antibodies)
IMMUCOR
Additional Materials Required: Test tubes (12x75 mm or 10x75 mm), pipettes,
isotonic saline or phosphate-buffered (approximately 15 mM) isotonic saline pH 6.5-
7.5, timer, centrifuge, and optical aid such as a hand lens or a concave mirror.
TEST METHOD:
1. In each of two test tubes, place a suitable volume of the serum or plasma
sample to be tested. The actual volume used will be dependent on the extent
of subsequent testing intended, but should be consistent with the ability to
obtain the recommended proportions of sample and substance. As an
example, ten drops of sample will suffice for limited testing, but 1 mL may
permit the sample to be tested against a panel of red blood cell suspensions.
2. Label the two tubes appropriately and add:
(a) to the first tube a volume of Gamma Pi Blood Group Substance equal to
one tenth the volume of sample in the tube (e.g. one drop of Pi
substance to 10 drops of sample, or 0.1 m L t o l mL of sample).
(b) to the second tube a similar appropriate volume of saline.
NOTE: The substance should be added directly to the sample, rather than
being allowed to run down the inside wall of the tube.
3. Mix the contents of both tubes thoroughly and allow to stand at room
temperature (23°±3°C) for at least 5 minutes. Incubating for greater than 5
minutes may enhance neutralization.
4. Mix the contents of both tubes again and proceed to test each substance-
treated and saline-diluted sample with Reagent Red Blood Cells in accordance
with the directions for use supplied with the particular product. Include at least
one red blood cell suspension known to have given a positive reaction when
tested previously with the original sample.
5. Read for agglutination in the prescribed manner and record test results.
QUALITY CONTROL: The test procedure provides the means to recognize the
effect of dilution resulting from the addition of one part of P i substance to 10 parts of
sample, by including a parallel test with sample similarly diluted by the addition of
saline. It is important that these parallel tests should be carried out on the same
specimen. In the event that both treated samples give negative reactions with red
blood cells observed previously to be agglutinated by the untreated sample, it is
if possible that the relatively slight dilution has adversely affected the reactivity of a
very weak antibody, or that the test has been performed under conditions that may
have differed in some small respect from those of the earlier test. For example, if the
Reagent Red Blood Cells used previously were still cold after recent removal from
the refrigerator, but have since warmed up to room temperature, the earlier test may
have been more sensitive for the detection of P i antibodies because of the lower
temperature of the test mixture. An alternative possibility, if a fresh specimen was
drawn for the test with P i substance, is that the reactions observed originally are not
capable of being demonstrated on the specimen drawn more recently.
As a quality control measure to demonstrate the continuing ability of this product to
neutralize P i antibodies, it is recommended that the test procedure should be
applied periodically to a serum known to contain anti-Pi (e.g. Gamma-clone® Anti-Pi
Blood Grouping Reagent).
INTERPRETATION OF TEST RESULTS: If P i + red blood cells are not agglutinated
by substance-treated sample, yet the saline-diluted sample still agglutinates P i + red
blood cells, the presence of anti-Pi is confirmed.
If substance-treated and saline-diluted samples show reactions of equal strength
and specificity with Reagent Red Blood Cells, the antibody present in the sample is
not capable of being neutralized with Gamma P i Blood Group Substance. In most
cases, the significance of this finding is that the antibody is not of anti-Pi specificity.
In rare instances, unusually potent examples of anti-Pi may not be neutralized (or
may be only partially neutralized) by the addition of substance in the amount
recommended. In such cases, significantly diminished reactions with Pi red blood
cells (e.g. a 1+ or greater difference in reaction strength between substance-treated
and saline-diluted sample) may be observed. If partial neutralization is suspected,
the test should be repeated using an increased ratio of substance to sample (e.g. 1
part of substance to 5 parts of serum or plasma, or even equal proportions of
substance and serum or plasma), together with a parallel test using the same
increased ratio of saline to serum or plasma.
In the case of antibody mixtures including anti-Pi antibody as a component, the
substance-treated sample will show the reaction pattern to be expected of the one
or more other antibodies in the mixture. The specific neutralization of the anti-Pi
component facilitates the identification of the other antibodies in the specimen by
enabling further tests to be performed without selecting Pi-negative red blood cells.
The test procedure may be applied to the reverse ABO grouping test in order to
eliminate discrepant reactions due to anti-Pi in the specimen being tested.
Assuming the absence of other antibodies reactive under the conditions of the
reverse grouping test, neutralization of the anti-Pi component enables anti-A and/or
anti-B in the specimen to provide proper confirmation of the results obtained in
forward grouping.

LIMITATIONS: As in all serological tests, such factors as contaminated materials,

improper incubation time, temperature, centrifugation, examination for agglutination
and deviation from the recommended test procedure, may give rise to false test
results.
In addition:
1 . Exceptionally potent examples of anti-Pi may only be partially neutralized by
the quantity of blood group substance recommended in the test procedure.
2. Specimens containing very weak antibodies may be diluted beyond the point of
reactivity by the addition of blood group substance in the proportion of 1 to 10.
3. In addition to neutralizing anti-Pi, this product will also neutralize anti-P k . This
antibody occurs regularly in combination with anti-Pi and anti-P in persons of
the Tj(a-) phenotype. Thus, in a specimen containing anti-P+Pi+Pk (anti-Tja ) the
anti-Pi and the anti-Pk would be neutralized, but the anti-P activity would not be
neutralized.

SPECIFIC P E R F O R M A N C E CHARACTERISTICS: Gamma Pi Blood Group

Substance is tested by the recommended test procedure and found to inhibit the
agglutination of P i + red blood cetts by a house standard antr-Pi serum having a titer
of at least 8. The product is shown not to neutralize other antibodies commonly
identified by the use of Reagent Red Blood Cells (such as those of the MNSs, Rh,
Lutheran, Lewis, Duffy and Kidd blood group systems). When stored correctly and
used in accordance with the procedure detailed in this direction insert, this reagent
will aid in the identification of unexpected blood group antibodies encountered
during antibody screening tests. The performance of this product is dependent on
adhering to the recommended methods found in this insert. For additional
information or for technical support, contact Immucor at 855-IMMUCOR (466-8267).

BIBLIOGRAPHY:
1. Frangois-Gerard CH, Brocteur J. Description of a Pi-like antigen present in
turtle-dove eggs. Abstract of a poster presented at XV Congress of the
International Society of Blood Transfusion, Paris 1978.

Insert code: 3004-4

Revised: 3/2017

Key:
Underline = Addition or significant change ▲ = Deletion of text