Blood Component Preparation Flashcards
What are 3 FDA requirements for Blood Component Preparation and Processing?
- Blood must be collected in sterile, pyrogen-free, uncolored, transparent containers.
- The blood bag must not have been entered before issuing for transfusion for any purpose, except to collect donor blood or when blood processing requires the use of a different container. (cannot be entered or spiked for any purpose except collection or making components)
- There should be no adverse effect on the safety of the product.
The purposes of anticoagulants and preservatives are
- Prevent blood from clotting
- Extend the life of RBCs
- Main collection bag will contain about 63mL anticoag/preservative
Blood bags are what?
closed systems with as many as four satellite bags attached
What three anticoagulants are commonly used in blood bags?
Citrate-phosphate-dextrose (CPD)
Citrate phosphate-2-dextrose (CP2D)
Citrate-phosphate-dextrose-adenine (CPDA)
What three additives commonly used in blood bags?
AS-1 (adsol)
AS-3 (nutricel)
AS-5 (optisol)
What is the outdate for blood anticoagulated with CPD?
21 days for Citrate-phosphate-dextrose (CPD)
What is the outdate for blood anticoagulated with CP2D?
21 days for Citrate-phosphate-dextrose (CP2D)
What is the outdate for blood anticoagulated with CPDA?
35 days for Citrate-phosphate-dextrose-adenine (CPDA)
Additives must be added to blood before when?
within 72 hours
but they are generally part of the closed bag system
Generally the additives function to?
Enhance RBC survival
improves flow of blood to pt during transfusion
Additive solution ingredients?
Dextrose Adenine Monobasic sodium phosphate Mannitol NaCl Sodium citrate citric acid
What are Rejuvenation solutions and how do they function?
Not routinely used
- can extend the expiration date
- contains pyruvate, inosine, phosphate, adenine
- but blood needs to be washed before use.
- Restores 2,3-DPG and ATP levels in RBC units during storage or up to 3 days after the unit has outdated
What are the criterion for measuring RBC viability?
75% of the RBCs in a transfused unit of RBCs must be in the recipient’s circulation 24 hours after transfusion
How is RBC viability measured?
measured by labeling RBCs with a radioisotope and then identified after transfusion
What is a storage lesions and what are its effects on a unit of blood?
A loss of RBC viability due to storage. -decreased pH -decreased Glu consumption -increase lactic acid -ATP decrease -2,3-DPG levels decrease giving RBCs and impaired capacity for delivering oxygen to the tissues -Reversible loss RBC function (may return to normal after TF)
Which component(s) can use additives to extend their life?
Only RBCs
What is the significance of a storage lesion?
It causes problems for neonates. Fine for adults
RBCs<7 days old best for babies
Which blood component has the highest risk for TACO?
Whole Blood
What are the best uses of whole blood?
exchange transfusion or massive blood loss in trauma
Whole Blood required Hematocrit?
33%
Whole Blood
- is not separated into components
- labile coag factors diminish on storage beginning after the first 24 hours
What anticoagulants/ preservatives are used for collection of whole blood?
ACD
CPD-21 days
CPD2-21 days
CPDA1-35 days
How is the RBC component made?
- Made by centrifuging whole blood
- plasma press aka plasma expresser forces plasma into a satellite bag
- Tubing btwn plasma and RBCs is sealed and thus sterility is maintained
What is the volume for RBC component?
225-350 mL
What us the expected hemoglobin and hematocrit of an RBC component?
HCT 65-80%
HGB 50-80 g/dL
any producted containing >2mLs of RBCS must undergo which test?
Must be crossmatched!
To make fresh frozen plasma, the product must be frozen within how many hours?
within 8 hours of collection
to store FFP for a year, what temp is required?
-18 degrees C
to store FFP for 7 years, what temp is required?
-65 degrees C
FFP can be processed further into?
Cryoprecipitate
How is Cryoprecipitate made?
Fresh Frozen Plasma is thawed at 1-6C Take out all plasma but 15mL cryo remains in original bag store at -18C outdate of 1 year
After cryoprecipitate is made from FFP what is the remaining plasma used for?
It can be processed into albumin, etc. or it is used as FFP, cryoprecipitate reduced.
What is the process for making Platelets from WB?
WB is soft spun to separate RBCs from PLTS/Plasma
PLTS/Plasma is hard spun
PLTS and 50ML are removed and stored at Room Temp with agitation for 5 days
What is the process for making FFP from WB?
WB is soft spun to separate RBCs from PLTS/Plasma
PLTS/Plasma is hard spun
PLTS and 50ML are removed
If processed within 8 hours, the product can be frozen as fresh frozen plasma
If processed after 8 hours, FP
If processed after 24 hours, Frozen Plasma 24
For FP and FP 24 there is a slight decrease in Factor 8, but the outdates are the same as for FFP
When are additives added to the RBCs during production?
After the soft spin of WB that separates RBCs from PLTs/Plasma
After additives, what further step is required for RBC processing?
Leukocyte reduction by filter with gravity
then RBCs are sealed and done.
What are 4 product variations of RBCs components?
must be marked on product container
Low volume RBCs
Irradiated RBCs
Leukocyte reduced RBCs (Aka Leukopoor)
Apheresis RBCs
What are low volume RBCs?
less than the normal amount of blood is collected at donation
Only the RBCs can be used
What are Irradiated RBCs?
For the prevention of graft vs. host disease
Outdate is changed to 28 days from time of collection or original outdate, whichever is shortest
1-6 C storage
increases K+ and decreases 2,3 DPG
Plts stay at RT
What are Leukocyted reduced RBCs aka Leukopoor)_?
- Accomplished by filtration either at blood donation center or at the patient bedside at infusion, w/n 72 hours
- outdate is unchanged
- Helps to prevent febrile TF reactions
- Decreased CMV TF doesnt reduce Graft vs. host disease
What are apheresis RBCs?
only RBCs are collected
All other components are returned to the donor
>/= 60 g/dL HGB per unit
How much coag factors, and antithrombin does FFP contain?
A normal amount
If FFP is leukocyte reduced, how does it affect the concentration of Factors?
Factors V, VIII, IX, XI, and XII are reduced
What is Thawed Plasma and what are its Factor levels?
Relabeled FFP 24 hours after thawed
Reduced levels of Factor V and VIII (labile factors)
5 day outdate
How is plasma thawed?
in a 37C water bath with plastic overlay to protect ports
What is PF24, what is its shelflife, and what happens if it is held for more than 24 hours?
Plasma frozen within 24 hours of collection
Once thawed has a shelf life of 24 hours at 1-6C
If held for 24hrs, must be labeled as thawed plasma and can be kept at 1-6C for four more days
What are all the names, configurations for Thawed Plasma?
FFP, PF24, or PR24RT24 plasma that has been held at 1-6 degrees for >24hours
How long is Thawed Plasma safe to use?
It can be held for up to 4 days after the initial 24 hour post-thaw period is up
What are all of the various configurations of plasma products?
FFP PF24 PF24RT24 Thawed plasma Plasma, Cryoprecipitate reduced Liquid plasma Recovered plasma Pathogen-reduced plasma
What is Plasma, Cryoprecipitate Reduced?
By product of cryoprecipitate production
Must be refrozen after cryoprecipitate production within 24hours at =-18C
Contains normal levels of Factor V and still has fibrinogen level of about 200 mg/dL
What is Liquid Plasma?
Plasma from WB
It can be separated from WB at any time during storage and stored at 1-6 C up to 5 days after the WB’s expiration
What is recovered plasma?
Plasma for manufacture
It can be shipped to fractioner and processed into derivatives such albumin or immune globulins
What is pathogen-reduced plasma?
Treated to inactivate microbes–doesn’t guarantee germ-free
Available in Europe and soon in the US
Txed with methylene blue, psoralen, riboflavin, and solvent/detergents
What plasma products can be collected by Apheresis?
FFP
PF24
PF24RT24
What is FFP?
held at 1-6 C
Frozen within 8 hours of collection
It is source plasma for manufacturing into fractionated components.
What is PF 24?
held at 1-6 C
Frozen within 24 hours of collection
What is PF24RT24?
Held at room temperature
frozen with in 24 hours of collection
Plasmapheresis frequency, donor management, RBC losses allowed?
- Serial plasmapheresis for source plasma can be performed up to two times per week
- For infrequent plasmapheresis, Donor selection and monitoring is the same as for WB
- RBC losses occur with the procedures, no more than 250 mL may be lost over 4-8 weeks
AABB requires cryoprecipitate to have what concentrations of Factor 8 and Fibrinogen?
> 80 IU of Factor 8 per unit
150mg Fibrinogen per unit
and it should contain Fibronectin and Von Willenbrand’s Factor
Cryoprecipitate production, storage, thawing, Use?
Prepared for FFP thawed at 1-6 C (the cold, insoluble protein precipitates at this temperature)
Thawed for use in 37 C water bath
Given in pools of 6-8 units for the average adult
Given in single units for babies/kids
Once a component has been thawed, can it ever be refrozen?
No.
How long can thawed Cryo be held for use in an open system, or in a single unit (Closed system)?
Pooled in open system, it can be held at room temperature for four hours after thawed.
As a single unit, it can be held at RT for 6 hours.
Pooling cryo involves rinsing each bag with?
sterile saline
What are the two methods of preparing platelets?
- Derived from WB=random donor plts
- They are resuspended in about 40-70 mL of plasma
- can be leukocyte reduced - Platelet Pheresis=single donor plts
- plts are removed by pheresis with other componenets returned to the donor
- resuspended in plasma from the donor
- can be leukocyte reduced
How are Plt pheresis PLTs and Random donor plts stored?
Both are stored at 20-24 C and must be constantly agitated to promote oxygenation, stabilize the pH and prohibit clumping
What does agitation of plts do?
maintains PLT metabolism and adequate in vivo recovery
Plt Transport container
keeps them at room temperature, but not aggitated
How many PLTs are required for a unit made of random donor derived PLTS?
5.5 x 10^10 PLTS
resuspended in 40-70 mL Plasma
A PLT unit must contain less than X RBCs
<2.4 x 10^9 RBCS
How often can plts be donated?
2x in 7 days or 24 times in 12 months
if RBCs cannot be returned, then defer for 8 weeks
How can risk of infection be reduced with PLT units?
Pathogen Inactivation
Visual inspection of PLTS
Can Plts be Leukoreduced?
Yes
How are PLTS given to adults?
Generally given in a pool of 4-6 units within 4 hours of storing
What are granulocyte packs used for?
Use has been controversial, studies have been inconclusive
Used for septic patients with WBC counts <500 /uL, whose bone marrow will eventually compensate, and whose infection has not responded to antibiotic therapy
AABB requires that granulocyte packs contain?
75% of granulocyte components contain at least a 1 x 10^10 granulocytes
Granulocyte dosage for Adults?
one pack
Granulocyte dosage for infants/children?
10-15 mg/kg body weight
What drugs are given to granulocyte donors and why are they given?
Drugs or sedimenting agents to increase WBC yields Hydroxyethyl starch (HES)--increases sedimentation of RBCs Corticosteroids -doubles the # of circulating granulocytes by mobilizing the ----pool Growth Factors (hematopoietic GF)
What are side effects of HES?
HAs
peripheral edema
What are the side effects of Corticosteroids?
HTN
DM
cataracts
peptic ulcers
If 2 mL of RBCs make it into the granulocytes?
then the product needs to be ABO/Rh compatible and crossmatched
Blood components are regulated by the FDA which requires medication manufacturers to verify the suitability of every raw material in their products.
What is the raw material in blood products?
Donors are the raw material the requires scrutiny.
How is donor suitability verified?
A sample of blood is taken and tested with every donation.
Testing includes:
ABO/Rh, including weak D testing for D negative
donors
Antibody screening
Infectious disease screening
What are the infectious disease tests routinely run?
HBV HCV HIV1/2 HTLV-I/II Syphilis Trypanosoma cruzi WNV ZIKA
What specific tests are run to test for HBV?
- Hepatitis B surface antigen
- Total (IgM and IgG) ab to hepatitis B core antigen
- HBV DNA
What specific tests are run to test for HCV?
- IgG antibody to HCV peptides and recombinant proteins
2. HCV RNA
What specific tests are run to test for HIV1/2?
IgM and IgG abs to HIV 1/2
HIV-1 RNA
What specific tests are run to test for HTLV-I/II?
IgG ab to HTLV-I/II
What specific tests are run to test for Syphilis?
IgG or IgG and IgM ab to T. pallidum antigens
Non-treponemal serologic test for syphilus (eg. RPR)
What specific tests are run to test for Trypanosoma cruzi
IgG antibody to T. cruzi
What specific tests are run to test for WNV?
WNA RNA
What specific tests are run to test for ZIKV?
ZIKV RNA
What screening test method is used for HBV?
- ChLIA or EIA
- ChLIA or EIA
- TMA or PCR
What screening test method is used for HCV?
- ChLIA or EIA or
2. TMA or PCR
What screening test method is used for HIV 1/2?
ChLIA or EIA
What screening test method is used for HTLV-I/II?
ChLIA or EIA
What screening test method is used for Syphilis?
Microahemagglutination or EIA
What screening test method is used for Trypanosoma cruzi?
ChLIA or EIA
What screening test method is used for WNV?
TMA or PCR
What screening test method is used for ZIKV?
TMA or PCR
What supplemental assays are used for HBV?
Positive HBV DNA
Neutralization
What supplemental assays are used for HCV?
Positive HCV RNA
What supplemental assays are used for HIV 1/2?
Positive HIV-1 RNA
HIV-1: Western blot
HIV-2: EIA
What supplemental assays are used for HTLV-I/II?
Western Blot
What supplemental assays are used for Syphilis?
Second FDA-cleared T. pallidum screening test
Infectious disease testing logistics required by FDA?
- Laboratories that perform donor tested are mandated by the FDA as biologics manufacturers
- Testing platforms must be licensed by the FDA
Infectious Disease serologic testing
test for antibody or antigen
enzyme immunoassay or chemluminescent immunoassay
Designed to be extremly sensitive
Nucleic acid testing of infectious diseases
Implemented to reduce the seronegative window periods
No repeat testing on reactive samples
HIV testing includes and what is the window period?
- Interview
- NAT testing for HIV RNA
- Serologic testing for HIV antibodies to detect IgM and IgG antibodies to HIV 1
- Estimated window period is 9 days
Hep B testing includes and what is the window period?
NAT
HBsAg
Anti-HBc, IgM, IgG
Window period is uncertain
HIV testing includes what and what is the window period?
- Interview
- NAT testing for HIV RNA
- Serologic testing for HIV antibodies to detect IgM and IgG antibodies to HIV 1
- Estimated window period is 9 days
Hep B testing includes what and what is the window period?
NAT
HBsAg
Anti-HBc, IgM, IgG
Window period is uncertain
Hep C testing includes what and what is the window period?
NAT for HCV RNA
Serologic testing for antibodies to HCV
Average window period: 7.4 days
HTLV I and II testing includes what and what is the window period?
- In US, found in immigrants and drug users
- infect lymphocytes
- serologic testing for antibodies to HTLV I/II
- Units associated with a reactive screening test cannot be released for transfusion
- testing has high false positive rate
- takes 2x reactio on subsequent donations
Syphiliis testing includes what and what is the window period?
- RPR
- Automated testing for T. pallidum antibodies
- Positive tests defer the donor for 12 months
- last TF associated case was 40 years ago
What is the risk of a bacterial contaminated blood product?
usu found in 1/3000 units
What are the sources of bacterial infected units?
bacteria on donor skin
bacteremia
multiplication during storage
What practices attempt to eliminate infection from donor units?
- Donor skin is carefully disinfected
- 10-40 mL of blood is initially diverted away from the main bag into the sampling pouch
- Visually inspect units before they are released
- rapid immunoassay tests for bacterial of PLTS
Risk of a prion in a blood product?
1 in a million
What is a prion?
- cause transmissable spongiform encephalopathies (TSEs)
- Transmitted by the implantation of products derive from infected CNS tissue
- No screening tests at this time; donors are excluded through interview/questioning
What blood-borne infections are transmitted by insect vectors?
Malaria West Nile virus T. cruzi Zika Babesia
How is malaria tested for?
- No good testing for it
- Rely on questionaire
- Donors who travel to malaria-endemic areas without prophylactic medications are deferred for 12 months
How is WNV tested for?
NAT for WNV
lipid enveloped virus
birds
How is T. cruzi tested for?
EIA for antibodies to T. cruzi
test for every donor
immunocompromised people can’t handle his disease of course
Chaga’s disease
How is T. cruzi transmitted ?
by food/drink
and from mother to child
How is Zika tested for?
there is no approved test, thus questionaire
How is Babesia tested for?
studies are being done regarding testing
permanent deferral for exposure, disease
Processing at the donor center includes
Bacterial testing of PLTS
Serology testing
Donor testing protocol
Nonreactive tests–considered negative are repeated negative x 2 = Negative
Positive on either test–>positive screen repeated in duplicate–>repeat positive–>confimatory test–>deferral and donor notified
(exception is syphilis testing)
What serology testing is performed at the donor center?
EIA or chemiluminescent assays NAT -HIV RNA -HCV RNA -HBV DNA -ID-NAT for WNV RNA
What is donor lookback?
performed whenever a repeat donor tests positive for an infectious disease test when they were previously negative
All donors with positive HIV and Hepatitis screens must have what next?
Confirmatory tests and if they are still positive, the donor is permanently deferred.
Positive testing for T. cruzi results in?
a permanent deferral
Cytomegalovirus in blood donors
Most blood donors have been exposed to this virus
CMV illness
- causes serious illness in immunocompromised individuals and low birthweight premies with CMV-mothers
- lipid-enveloped DNA virus in the Herpesviridae family
- causes lifelong infection, persists in a latent state
- Transmitted through blood transfusion–through intact WBCs and cellular components
- Most blood donors have been exposed to the infection
Commercial plasma derivatives
prepared from large pools of plasma obtained from thousands of donors
paid donations
Pathogen Inactivation
can remove and/or inactivate many of the pathogens and viral agents that have contaminated these products in the past with methods: Prolonged heat Solvent detergent (SD) treatment
Benefit and limitation of Pathogen Inactivation
- alternative to relying on donor questioning and testing to restrict infectious donations
- not routinely performed on donor blood, but is under study as a possibility
- Human parvovirus and fifth disease can’t make it through
Pathogen Inactivation for Plasma: commercially prepared pools
Solvent/detergent treatment
FDA cleared
Pathogen Inactivation for Plasma: individual units
Psoralen + UV light FDA cleared
Riboflavin + UV light Not cleared
Methylene blue + light Not cleared
Pathogen Inactivation for Platlets
Psoralen + UV light FDA cleared
Riboflavin + UV light Not cleared
UV light Not cleared
Pathogen Inactivation of RBCs/ WB
Amustaline and glutathione No
Riboflavin + UV light No
Quarantine in the blood center
All units of blood collected are immediately placed in quarantine in a designated area of the donor center while:
- records are reviewed
- comparison of records takes place
- deferrals are examined
- testing is completed
Where is TF unsuitable blood stored?
Away from and separated from the general blood storage
Before Blood is ready to be labeled, what four things must happen?
- holds must be resolved
- discrepencies must be resolved ex, name change
- Infectious disease testing must be complete
- all QC must be satisfactory
