blood transfusion lab Flashcards
what are antigens?
part of the surface of cells
all blood cells have what?
antigens
what are antibodies?
protein molecules –usually of the immunoglobulin classes: IgG and IgM
when do reactions to blood usually occur?
when the antibody in the plasma reacts with an antigen on the cells
where are antibodies found?
in the plasma
when are antibodies produced?
produced by the immune system following exposure to a foreign antigen
how many known blood group systems are there?
26
-ABO and Rh are clinically most important
antigens in transfused blood can stimulate what?
can stimulate a patient to produce an antibody, but ONLY if the patient lacks the antigen themselves
is the frequency of antibody production high or low?
very low but increases the more transfusions that are given
when is antibody production stimulated?
Blood transfusion
-i.e. blood carrying antigens foreign to the patient
Pregnancy
-fetal antigen entering maternal circulation during pregnancy or at birth
Environmental factors
-(i.e. naturally acquired e.g. anti-A and anti-B)
where do Antibody – Antigen reactions occur?
in vivo (in the body)
- leads to the destruction of the cell either:
- directly when the cell breaks up in the blood stream (intravascular)
- indirectly when liver and spleen remove antibody coated cells (extravascular)
in vitro (in the laboratory) -reactions are normally seen as agglutination tests
define agglutination
clumping together of red cells into visible agglutinates due to antigen-antibody reactions (antibody cross-linking with the antigens)
what is specific?
the antigen-antibody reaction agglutination can identify:-
what can agglutination identify?
The presence of a red cell antigen
-i.e. blood grouping
The presence of an antibody in the plasma
-i.e. antibody screening/identification
55% of the UK have which antigens?
A and B antigens
97% UK have which anti bodies?
Anti-A, anti-B or anti-A,B antibodies
what is high risk?
A or B cells being transfused into someone with the antibody in a random situation
ABO antibodies can activate what?
complement
-causing INTRAVASCULAR HAEMOLYSIS
almost all serious/fatal transfusion reactions caused by technical/clerical error are due to what?
ABO incompatibility
blood type phenotypes, antigens and antibodies
groups A, B, AB, O
antigens A, B, A + B, no antigens
antibodies anti-B, anti-A, anti-A and anti-B, none
what happens in a blood grouping test?
patient’s red cells and plasma are both tested
- Test patient’s red cells with anti-A, anti-B and anti-D
- agglutination shows that a particular antigen is on the red cells
- no agglutination shows the antigen is absent - Test patient’s plasma with A cells and B cells
- agglutination shows that a particular antibody is in the plasma or serum
- no agglutination shows the antibody is absent
ABO Compatibility
- who can O blood types donate to?
- who can A blood types donate to?
- who can B blood types donate to?
- who can AB blood types donate to?
- O, A, B, AB
- A, AB
- B, AB
- AB
other than the ABO grouping system, what is other blood grouping system in the body?
Rh grouping system
- RBC’s sometimes have another antigen, a protein known as the RhD antigen
- If this is present, your blood group is RhD positive
- If it’s absent, your blood group is RhD negative
what is the most important antigen in the Rh grouping system?
D
people with the D antigen are what?
D positive (85% of UK)
people who don’t produce any D antigen are what?
D negative (15%)
name the antigens involved in the Rh grouping system
antigens: D, C, c, E and e
what typing is most important after ABO and how does it work?
Rh (D) typing
- must be tested in duplicate or tested each time and compared to historical result
- patient / donor classified as D pos or D neg
Clinical significance of Rh
-Transfusion
- D antigen is very immunogenic and anti-D is easily stimulated - PREVENTION!
- All Rh antibodies are capable of causing severe transfusion reaction- ANTIBODY
Clinical significance of Rh
-Pregnancy
- Rh antibodies are usually IgG and can cause haemolytic disease of the newborn
- Anti-D is still most common cause of severe HDN
what occurs in Haemolytic disease of the Newborn (HDN)
Rh- mother carries a Rh+ foetus. Rh antigens can from the foetus can enter the mothers blood during delivery. In response to the foetal Rh antigens, the mother produces Anti-Rh antibodies. If the mother becomes pregnant with another Rh+ foetus, the anti-Rh antibodies could cross the placenta and damage foetal RBC’s.
HDN – laboratory testing
Blood group and antibody screen at antenatal booking to identify pregnancies at risk of HDN
D negative women who may need anti-D prophylaxis
Atypical antibodies are quantified periodically to assess their potential effect on the fetus
the scan at 28 weeks looks at what?
Blood group and antibody screen
how can HDN be prevented?
An injection of anti-D will bind to and remove any fetal D positive red cells in the circulation
1500 iu of anti-D is given routinely at 28 weeks and a smaller dose (usually 500 iu) after delivery if baby RhD+
In some hospitals 2 smaller (500 iu) doses are given at 28 and 34 weeks instead of the 1 larger dose
Anti-D is also given after any event that may cause a feto-maternal haemorrhage (bleed between mum and fetus) such as:
Abdominal trauma
Intrauterine death
Spontaneous or therapeutic abortion
why is Antibody Screening important?
its important we screen for these antibodies so that if detected, antigen negative blood can be provided to avoid causing a immune reaction
-prevent a haemolytic transfusion reaction
how does Antibody screening work?
Patients serum is mixed with 3 selected screening cells, incubated for 15 minutes at 37oc and then centrifuged for 5 minutes.
Any clinically significant antibodies reacting at body temp should be detected and then identified using panel of known phenotyped red cells.
Specific antigen negative blood can then be provided for these patients to avoid stimulating an immune response.
If an antibody is detected we must?
Identify the antibody
Assess its clinical significance
- For transfusion
- In pregnancy
How to Identify an antibody
Compare pattern of reactions with each reagent cell of ID panel with the pattern of antigens on the reagent cells
Matching pattern will identify the antibody
IgG vs IgM?
IgM antibodies can span the gap between RBCs
IgG can not, because too small to overcome ZETA potential (+ve charge)
how can IgG span the gap - what modification happens?
LISS (low ionic strength saline) is negatively charged, so neutralises positive ZETA potential
Therefore IgG can now span the gap.
Indirect anti-globulin test (IAT)
Used to detect IgG antibodies
LISS counteracts Zeta potential
Results in agglutination
Used for:
- Screening for antibodies
- Identifying antibodies
- Cross-matching donor blood with recipient plasma when there are known antibodies or a previous history of antibodies.
Cross-matching?
Immediate spin cross-match (ISX)
Full Indirect Antiglobulin test (IAT) cross-match
-only if Antibody screen positive or patient has known antibody history
Immediate spin cross-match (ISX)
basically checking the ABO blood group, and with IgM antibodies there is no ZETA potential, therefore no need to IAT
Antibody screen is negative
Checking donor red cells against patients plasma
- ABO check - Incubate for 2 – 5 minutes (room temp), spin and read
Full Indirect Antiglobulin test (IAT) cross-match
Antibody screen positive or patient has known antibody history
Select antigen negative donor red cells and incubate with patient serum for 15 minutes at 37oC
Blood donors
Only 4-6% of eligible population donate
Eligibility:
17 - 65 years old (first donation)
Over 50kg
Donor Selection
Questionnaire: lifestyle, health, not previously transfused
Collection procedure arm cleansing / diversion pouch
Comprehensive testing of all products -Viral: HIV 1+2 Hepatitis B Hepatitis C Syphilis HTLV
Platelets
Bacteria
ABO, RhD, K, antibody screen
Relative risks of transfusion for HIV?
1 in 7 million for HIV infection
how does transfusion of red cells work?
Concentrated red cells (packed cells) in a suspension of SAGM
-with symptomatic anaemia you can do an exchange transfusion
If significant bleeding anticipated, activate the major haemorrhage protocol
transfusion of Fresh Frozen Plasma?
FFP contains all clotting factors
Given for coagulopathy with associated bleeding
Requires clotting screens to monitor
Only has 24 hour life after thawing
(five days for major haemorrhage)
transfusion of Platelets?
Adult pool of platelets from 4 donors (suspended in plasma from 1 donor)
Platelets required to create clots to reduce bleeding
Some drugs given to reduce efficacy of platelets (anti-platelet agents) so patient history important
Cryoprecipitate
Contains Factor VIII, VWF and fibrinogen
2 units usually given at one time
Monitor fibrinogen levels by clotting screens
Haemovigilance
Serious Hazards of Transfusion (SHOT):
- Voluntary reporting
- Report all Serious adverse Events (SAE) and Serious adverse reactions (SAR)
Serious Adverse Blood reactions and events (SABRE):
-Mandatory reporting