Exam B Practicum SQ Flashcards
Enzymes are proteins that do what for chemical reactions?
They increase the rate of a chemical reaction
In transfusion medicine is what altered by enzymes?
RBC antigens
What are the two most commonly used enzymes in immunohematology?
Papain and ficin
What are the less commonly used enzymes in immunohematology?
bromelin, pronase, trypsin, and chemotrypsin
Where do destroyed antigens reside after enzymes have done their work?
On the part of the molecule that the enzyme removed from the RBC membrane.
When are enzymes used?
To clarify the reaction of weakly reactive antibodies or antibody that may be present or confirm the presence of an antibody.
What is the one step technique for enzyme treatment?
Enzyme is added as an enhancement to the tube containing test plasma and RBCs. The technique is carried out in IAT phase of testing.
What is the two step technique for enzyme treatment?
Treatment of RBCs with enzyme and washing prior to the addition of patient plasma. Method preferred b/c enzymes any antibodies present may be damaged.
How are untreated RBCs and enzyme treated RBCs tested?
They are tested together at the same time to identify all antibodies that may be present in the test sample.
Why would an MLS use neutralization in blood bank?
Aid in identifying a suspected antibody that does not react with a typical pattern. It is done by combining it with a soluble form of the corresponding antigen.
What does saliva neutralize?
ABO (secretor), and Lewis (secretor for Le^b)
What does hydatid cyst fluid and pigeon egg whites neutralize?
Neutralizes anti-P1 b/c it has P1 antigen in the fluid.
What antigen is neutralized by urine?
Sd^a
What is neutralized by serum?
Chido and Rodgers
What is neutralized by breast milk?
What is neutralized by amniotic fluid?
What antigens are destroyed by sulfhydryl reagents? Reagents such as 2-mercaptoethanol (2-ME), Dithiothreitol (DTT, and 2- aminoethylisothiouronium bromide (AET)
Kell antigens
Kidd antibodies can show ___A____ and are ___B___ by enzymes.
A. dosage / B enhanced
What is the purpose of doing an elution?
Freeing any antibodies that are bound to the membrane of RBCs using an acid.
What techniques can be combined with elution?
DAT to further investigate for possible antibodies. Adsorption to detect weakly reactive antigen such as weak ABO subgroup.
What methods are used to detect cold reactive antibodies or antibodies that have a broad temperature range?
Lansteiner and Miller heat elution
Weiner’s freeze–thaw method
Lui’s freeze–thaw method and sonication
What methods are suited for recovering warm- reactive alloantibodies, or autoantibodies?
Organic solvents such as ether, xylene, methylene chloride, and chloroform. Acids can also be used to recover the same antibodies such as digitonin, citric, or glycine.
What procedures remove antibodies but leave RBCs intact?
Gentle heat, chloroquine, and acid glycine - EDTA methods
What factors can cause an eluate to fail?
Improper technique
Inadequate stroma removal
Insufficient washing of the sensitized red blood cells before eluting the
antibody
Incorrect method of transferring washed red blood cells
Incorrect procedure used during the wash phase
Use of saline to prepare eluates
Adsorption is a technique used to….
Remove an antibody or antibodies from plasma. It is done to remove autoantibodies from a patient’s specimen in order to see if other antibodies are present in plasma.
What is the preferred method to remove autoantibodies?
Autologous adsorption. It cannot be used on patients that have been transfused within 3 months because donor RBCs may also have autoantibodies.
What is the preferred method for removing autoantibodies if a patient has been transfused within 3 months?
Allogenic adsorption. It can be used on patients that have been transfused. It can be paired with elution techniques. To separate mixtures of antibodies for identification.
How are RBCs chosen for alllogenic adsorption?
RBC antigens that stimulate the most clinically significant antibodies are considered.
What is the risk of allogenic adsorption?
Not detecting clinical significant antibodies if a weakly reactive alloantibody is diluted or antibody to a high frequency antigen is absorbed. Autoantibodies are only partially absorbed may mimic the reaction pattern of alloantibodies.
Rabbit erythrocyte stoma can remove…
Cold autoantibodies, anti-B, and antibodies in the P blood group system (anti-P1).
True or false, can plasma from adsorption with rabbit erythrocyte stroma be used for ABO typing?
No
When is rabbit erythrocyte stroma preferably used?
When an autoadsorption cannot be performed because a patient with a cold autoantibody has been recently transfused.
What is the risk with rabbit erythrocyte stroma?
It can complex or decrease the strength and remove other IgM antibodies including clinically significant antibodies.
What does titration do?
Used to measure the strength (concentration) of an antibody.
Describe titration
A serial dilutions are made of the antibody containing plasma and tested against selected red blood cells to determine the highest dilution causing a positive reaction. Result or titer is expressed as the reciprocal of the highest plasma dilution reacting +1.
What is titration score?
A number assigned to each positive reaction in the titration based on the strength of the reaction (+1-+4).
What does titration score tell the blood banker about the antibody?
Sum of these scores reflect the total binding strength of the antigen and antibody molecule (known as titration score or avidity). Its useful for comparing how an antibody reacts to different RBCs or how different antibodies react to the same RBC.
Complete the statement; Two antibodies can have the __A__ titer but __B__ titration scores, demonstrating the same strength but differing avidity.
A = same
B = different
Reliable titration results depend on…
technique
What situations is titration useful?
- Used for determining antibody activity
-Determines potential severity of HDFN in alloimmunized pregnant women
-Useful in identifying antibodies to certain high-incidence antigens
-Can distinguish characteristic high-titer, low-avidity antibody (HTLA) and a weakly reactive antibody with a low titer
What cannot be done in a blood panel if all cells are positive?
No antibodies can be excluded
If the autocontrol or DAT is negative and the patient has been transfused or pregnant then the reactions are likely due to….
alloantibodies! Note: Both combinations of multiple antibodies and a single antibody to a high-incidence antigen may cause this pattern of reactivity.
If the autocontrol or DAT is positive, the patient has not been transfused recent, then the results are likely due to an…
Autoantibody that could be warm reactive, cold reactive or both.
If the autocontrol or DAT is positive and the patient has been transfused recently, the results may indicate either….
an autoantibody or a delayed serological transfusion reaction
False-positive panreactivity may occur with….
rouleaux or antibodies to reagents
What is the typical pattern from a patient that has developed an antibody to a high incidence antigen?
Negative autocontrol or DAT, but almost all screen, panel, and donor RBCs react positively.
What is the best way to confirm the specificity of an antibody?
Testing with rare selected cells that lack the antigen.
What is an alternative to confirming an antibody if no rare selected cells are available?
Testing the patient’s plasma against treated cells may provide clues to the antibody’s identity. Know a patient’s ethnicity can be helpful too!
Warm autoantibodies may cause….
Autoimmune hemolytic anemia. They react with patient’s own RBCs at body temperature.
What is the typical pattern of warm autoantibodies?
- Positive autocontrol
-Positive DAT
-All screen, panel, and donor RBCs react positive
Describe panagglutination
An antibody that causes all cells tested to agglutinate. A panagglutinate can be a warm autoantibody (panagglutinin)
What method is used for a patient that may have an alloantibody and hasn’t been transfused in the last 3 months?
autoadsorption. Adsorb plasma and test for alloantibodies.
Patients with warm autoantibodies seem to have an increased….
sensitivity to alloimmunization compared to other patient populations.
Patient cells can be ________ for antigens of clinical significance.
Phenotyped. The cells may be treated to remove antibody prior to phenotyping if the antiserum requires an IAT phase.
What may not be necessary if the patient has phenotypically matched RBC donor units available?
Allogenic adsorption
What kind of antibodies react at temperatures between 4C and 25C and can be autoantibodies or alloantibodies?
Cold agglutinins
What technique or method differentiates cold agglutinins from other antibodies?
autocontrol or DAT
Why are cold agglutinins a concern?
They create problems in the lab by interfering with ABO typing and mask reactions of more clinically significant antibodies.
What are the most common cold autoantibodies?
autoanti-I (big i)
Cold autoantibodies can be specific to what antigens?
Big i, little i, IT, IH, and Pr
What is a good way to show if cold-reactive autoantibodies are clinically significant?
Antibody titer (serial dilution) and thermal amplitude
If cold autoantibodies are masking significant alloantibodies, how can cold autoantibodies be removed?
autoadsorption using a low temperature or by adsorption with rabbit erythrocyte stroma
List some cold alloantibodies
Anti-M, anti-N, anti-P1, anti-Pk, anti-Lea, anti-leb, and anti-Lua
What are three ways to avoid detecting cold alloantibodies using tube method?
- Eliminate IS phase
- Eliminate microscopic readings
- Testing with anti-IgG instead of polyspecific antihuman globulin.
What is the pre-warm technique?
Used to detect presence of slinically significant antibodies in the presence of cold-reactive antibodies.
What is the risk of the prewarm technique?
Warming the plasma causes a decrease in the reactivity of 40-47% of clinically significant antibodies per research.
What does a lab scientist do in the prewarm technique?
Plasma and cells are warmed separately at 47C prior to testing in an attempt to avoid binding of cold antibodies.
If performing tube technique, warm saline may be used in the wash step of IAT.
Prewarming can avoid detecting cold reactive antibodies it could resolve ….
ABO descrepancies due to cold antibodies
What can the lab scientist do to predict the potential complications of cold agglutinins during cardiac surgery?
thermal amplitude and titers
Cold agglutinins are present and a problem in cardiac surgery. What can the surgeon do to avoid disaster?
Blood or patient’s body temp. kept above temp at which cold antibodies react or a plasma exchange to remove cold agglutinins prior to surgery.