PROCEDURE CONTINUED:

f. Resuspend the cells and examine macroscopically for agglutination. Grade the
reactions as 1+, 2+, 3+, 4+ and 0 for negative. Record results in Transfusion
Service Record Book.

INTERPRETATION:

1. Agglutination of red blood cells indicates the presence of Anti –A1 subgroup.
2. No agglutination of red blood cells indicates the presence of Anti-A2 subgroup,
and weak subgroups of A.
3. Positive and negative controls must be run simultaneously to be a valid test.

LIMITATIONS:

1. Do not incubate the test tubes at 37 degrees centigrade, as this may produce false
positive agglutination.

2. False positive tests may result from the presence of bacterial or chemical
contamination of serum, cells, or saline; extended or improper incubation and/or
centrifugation.

3. False negative tests may result from improper or prolonged storage of cells,
improper collection or preparation of saliva, inadequate incubation, improper cell
concentration, and/or centrifugation.

NOTE: ANTIGEN NEGATIVE UNITS MUST BE GIVEN TO PATIENT’S WITH

CORRESPONDING ANTIBODIES PRESENT. EXCEPTIONS ARE
ANTI H and ANTI I.
ANTI - s AND ANTI – S

1. Label tube for cell suspension with patient last name and first name.
2. Prepare a 5% saline cell suspension of washed red blood cells.
3. Add one (1) drop of Immucor Anti – S or Anti – s to appropriately labeled tube.
4. Add one (1) drop of cell suspension to appropriately labeled tube
5. Mix well.
6. Incubate at 36 – 38 C for 15 – 20 minutes
7. Wash three times with saline, making certain to decant last wash completely.
8. Add tow drops of Immucor Anti –IgG - C3d Anti Human Globulin
9. Centrifuge for 15 seconds. Resuspend the cells and examine macroscopically
For agglutination. Grade the reaction as 1+, 2+, 3+,4+, and 0 for negative.
Record results in Transfusion Service Record Book
10. Add one (1) drop of Immucor Checkcell (IgG Coated Antiglobulin) to all negative
tubes. The check cells must check for test to be valid.

INTERPRETATION

1. Agglutination of red cells is a positive test result and indicates the presence of S
And little s antigens.
2. No agglutination of red cells is a negative test result and indicates the absence of
S and little s antigens
2. Positive, Negative and patient controls must be run simultaneously. If patients
control shows, agglutination test is not valid.

LIMITATIONS

1. False positive or false negative test results can occur from bacterial or chemical
contamination of test materials, inadequate incubation time and temperature
improper centrifugation, improper storage of materials or omission of test
reagents.
2. Red cells demonstrating a positive direct Antiglobulin test cannot be accurately
tested for the S or little s antigens with these reagents designed for use at the
Antiglobulin Phase of testing.
3. Certain antigens (Bg, SdA) can be present in an exalted state on red blood cells.
This phenomena may be a source of rare false positive reactions which may occur
with more than one lot of given specificity.
4. Suppressed or diminished expression of certain blood group antigen may
conversely give rise to false – negative reactions. For these reactions, caution
should always be exercised when assigning genotypes on the basis of serologic
test results.
5. Red cells from individuals with the unusual MN System Phenotypes may produce
reactions with Anti – S and anti – s that are considerably weaker than those of
control cells tested in parallel.
ANTI - S AND ANTI – s

Results are reported in the Sunquest Computer in the following format:

1. Type in your access code which is your initials

2. Type in your password
3. At function: type in BOP
4. At BOP type in the hospital number or accession number
5. In BOP at option press return and proceed to gird
6. In window of addition tests press key insert to come off keyboard. Press the
semicolon key, the Sunquest computer will allow you to add a test.
7. Patient typing antigen, add AGI
8. In the additional test window at AGI, press the semicolon key to come back off
keyboard.
9. Add code for antigen being tested
10. Unit antigen testing is done in transfusion status window
11. Follow the same procedure as patient antigen testing. The code is AO
12. Press shift *
13. Using caution verify the result is correct. Type A to accept the results M to
modify the results or R to reject the results.

REFERENCES:

WIDDMAN, Frances K. The American Association of Blood Banks Technical Manual,

13th Ed., 1999.
Package insert Immucor Anti Lewis A and Anti Lewis B
Package insert Immucor Anti M and Anti N
Package insert Immucor Anti Cw
Package insert Immucor Anti Jka and Anti Jkb
Package insert Immucor Anti Fya and Anti Fyb
Package insert Immucor Anti A1 Lectin
Package insert Immucor Anti s and Anti S
Package insert Immucor Anti c and Anti C
Package insert Immucor Anti e and Anti E