1.
HDFN Test (Kleihauer-Betke Test)
Principle: The test is based on the principle that red cells containing fetal hemoglobin
(HbF) are less susceptible to acid elution than cells containing adult hemoglobin (HbA).
Procedure:
a. Make a blood smear using maternal blood and treat with acid.
Fetal cells will remain intact and appear pink.
Mother’s cells become ghost cells.
b. Count 2000 cells then calculate:
%fetal cells= (#fetal cells/2000) x 100
c. Compute for the volume of fetomaternal hemorrhage:
%fetal cells x 50 ml= volume of FMH
d. Compute for the number of vials:
(Volume of FMH/30) + 1= number of vials needed for RhIG
Reference:
Harmening, D. M. (2012). Modern Blood Banking & Transfusion Practices (6th ed.). pp.
434
2. Antibody Panel Testing/Screening
Principle: The antibody screen test is used to detect clinically significant unexpected
antibodies in both the blood donor and the intended recipient as part of pretransfusion
compatibility testing.
Procedure (Tube Method):
Reference:
Harmening, D. M. (2012). Modern Blood Banking & Transfusion Practices (6th ed.). pp.
217,218
3. Apheresis
Principle: The main principle behind apheresis is the separation of donor blood into its
component parts by either centrifugation or membrane filtration.
Procedure:
a. Blood is removed from an individual (usually with a large-bore needle).
b. Mixed with an anticoagulant.
c. Transported directly to the separation device (typically a machine with a
centrifuge bowl or belt).
Separated into specific components.
d. Once the components have been separated, any component can be withdrawn.
e. The remaining portions of the blood are then mixed and returned to the donor or
patient.
Reference:
Harmening, D. M. (2012). Modern Blood Banking & Transfusion Practices (6th ed.). pp.
332
4. Component Therapy
Principle: Transfusion of the specific component needed by the patient.
Blood component procedures:
a. Whole Blood (WB)
Preparation: Approximately 450 mL of blood with anticoagulant (CPD or
CPDA-1)
Shelf life: CPD- 21 days at 1-6°C; CPDA-1: 35 days
Dilution: 1:8
A unit of blood must be transfused within 24 hours if the seal on the bag is
broken to remove plasma.
b. Packed Red Blood Cells
Preparation: Each unit of packed red blood cells contains approximately
250 mL.
▫ Prepared by removing 200 to 250 mL of plasma from a unit of WB.
Shelf life: Cells prepared in an open system must be transfused within 24
hours.
▫ If cells are prepared using the close system, they have the same
expiration date as the original unit of the WB.
c. WBC poor RBC
Preparation: 70% of the original WBCs removed and at least 70% of the
original RBCs are left.
▫ Methods of obtaining leukocyte poor RBCs:
Centrifugation, filtration, and washing.
Shelf life: Closed system- same as the original unit of blood.
▫ Open system: 24 hours
▫ Stored at 1-6°C
d. Washed RBCs
Preparation: Plasma is removed from WB after centrifugation.
Washing: Removes plasma proteins.
Shelf life: Washed RBCs have a shelf life of 24 hours after the original unit
is opened. Stored at 1-6°C.
e. Frozen Deglycerolized RBC
Preparation: RBCs to be frozen are collected in CPD, CPDA-1. Should be
frozen within 6 hours.
Deglycerolization:
▫ Begins with thawing the cells at 37°C, then washing multiple time in
a gradient concentration of saline, beginning with hypertonic
concentrations and ending with an isotonic solution containing
glucose.
▫ One unit of deglycerolyzed RBCs contain approximately 180 mL of
cells.
Shelf life: Stored between 1-6°C and must be transferred within 2 hours of
deglycerolization.
Frozen with the use of glycerol
▫ High glycerol (40% wt. per volume)
Freezer temperature
-65°C
▫ Low glycerol (20% wt. per volume)
Freezer temperature
-120°C
f. Platelet Concentrate
Preparation: Platelet-rich plasma (PRP) is separated at room temperature
by centrifugation from RBCs within 6 hours of collection of WB.
▫ The PRP is then centrifuged, and the resulting platelet-poor plasma
(PPP) supernatant is removed, which leaves approximately 50 mL
of plasma with the platelet concentrate.
Shelf life: Platelets are stored at RT with continuous gentle agitation.
g. Granulocyte concentrate
Preparation: May be prepared by leukapheresis or from a freshly drawn
donor unit.
Administer corticosteroids to the donor 12-24 hours prior
▫ Increase the number of circulating granulocytes by pulling them
from the marginating pool.
Shelf life: 24 hours after separation at RT.
▫ Granulocytes should be transfused ASAP because their half-life is
only 6 hours.
h. Fresh Frozen Plasma
Preparation:
▫ Must be frozen within 8 hours of collection.
▫ Plasma is immediately frozen at or below -18°C.
Shelf life:
▫ Plasma should be stored at or below -18°C.
▫ It should be thawed at 37°C and transfused within 24 hours of
thawing.
▫ Thawed FFP should be stored between 1-6°C if it is not transfused
immediately.
i. Plasma Derivatives
Separated from WB at any time during the unit’s shelf life up to 5 days
after the expiration date.
Plasma may be pooled, purified or fractioned into albumin or plasma
protein fraction.
Shelf life: Plasma derivatives have a shelf life of 5 years at 1-6°C.
j. Cryoprecipitate
AABB: 150 mg of fibrinogen for each unit of cryoprecipitate
Component preparation: 5 units are pooled
Each pool: 750-1250 mf of fibrinogen
k. Cryoprecipitated Anti-Hemophilic Factor
Prepared from FFP thawed slowly between 1-6°C
Each unit: Contains at least 80 units of Factor VIII
l. Factor VIII Concentrate
From fractionation and lyophilization of pooled plasma.
Stored at refrigerator temperature and is reconstituted with saline
m. Factor IX concentrate
Prepared from pooled plasma using various methods of separation and
viral inactivation.
Reference:
Harmening, D. M. (2012). Modern Blood Banking & Transfusion Practices (6th ed.). pp.
312-319
