PRE TRANSFUSION

TESTING
CHAPTER 10
Pretransfusion testing
Series of serological and non-serological protocols and
testing procedures with ultimate objective of
preventing immune mediated and hemolytic
transfusion reaction
Not guarantee normal survival of transfused RBCs in
the recipient circulation
But there should be no significant destruction of the
recipient’s own RBC
STEPS IN PRETRANSFUSION
TESTING
REQUEST FOR TRANSFUSION
IDENTIFICATION OF RECPIENT AND BLOOD
SPECIMEN COLLECTION
TESTING OF RECIPIENT’S BLOOD SPECIMEN
DONOR RBC UNIT TESTING
DONOR RED CELL UNIT SELECTION
COMPATIBILITY TESTING
LABELING OF BLOOD AND BLOOD COMPONENTS
WITH THE RECIPIENT’S IDENTIFYING INFORMATION
AND ISSUE
REQUEST FOR TRANSFUSION

NEED AN ACTUAL REQUEST FOR BLOOD

TRANSFUSION
CHECK FOR COMPLETE INFORMATION ABOUT THE
PATIENT
NAME , PATIENT ID NUMBER, SEX, AGE,
BIRTHDAY (OPTIONAL), DIAGNOSIS, WRITTEN
REQUEST FROM THE PHYSICIAN, SIGNATURE OF
PHYSICIAN , BLOOD TYPE OF THE PATIENT
(OPTIONAL), REQUIRED BLOOD COMPONENT
POSITIVE IDENTIFICATION OF
RECIPIENT
Clerical error- major cause of transfusion associated fatalities
Misidentification of the recipient- most common error
ID wristband comparison to the request-recipient full name
and unique identification number
Printing must be legible
Any discrepancies must be completely resolved before the
sample is taken
Nameplates on the wall or bed label must never be used to
verify identity
If no wristband a positive identification must be attached
before collecting the sample
POSITIVE IDENTIFICATION
If no wrist band – coherent – ask the patient to state his full
name; ask the patient birth date of present in requisition form
Never asked “Are you Juan dela Cruz?”
If patient is young or incoherent- ask reliable professional
individual
COLLECTION OF PATIENT
SAMPLES
Proper collection technique- avoid hemolysis
Hemolysed sample cannot be used in pretransfusion testing
If unavoidable- observe the extent of hemolysis and if it
increases during stage of testing
Serum instead of plasma- fibrin in plasma can be distinguish as
an agglutination
Plasma anticoagulant may inactivate complement activation
some antibodies may not be detected
Tubes must be labelled before leaving the bedside of the patient
BLOOD COLLECTION
TO AVOID CONTAMINATION, COLLECT BLOOD FROM THE ARM
WITHOUT IV LINES, IF NOT POSSIBLE BELOW THE INFUSION
SITE NOT ABOVE. IF FROM THE IV LINE STOP THE FLOW FOR 5
TO 10 MINUTES , DISCARD THE FIRST 10 ML, THEN OBTAIN THE
SAMPLE
IF THE SPECIMEN IS RECEIVED IN THE LABORATORY, CHECK ALL
THE NECESSARY INFORMATION FROM THE TUBE AND THE
REQUEST
RECIPIENT SAMPLE SHOULD BE TESTED AS SOON AS POSSIBLE,
IF CANNOT BE PERFORMED IMMEDIATELY KEEP THE SAMPLE
TO 1° C TO 6 °C; washing of patient RBC is important
FOR BETTER HUMORAL IMMUNE RESPONSE SAMPLE TO BE
USED MUST BE FEWER THAN 72 HOURS AFTER COLLECTION
COLLECTION OF DONOR SAMPLE
Must be collected at the same time with the recipient
Pilot sample may be clotted or anticoagulated
RBC for pretransfusion testing can be prepared from
segmented tubing
Put a hole in the segment using a lancet – self sealing to
prevent contamination
Cut the segment
Both recipient and donor sample must be stored for a minimum
of 7 days following transfusion (stoppered,
carefully labelled and refrigerated at 1-6°C)
COMPATIBILITY TESTING
PROTOCOLS
Testing the Donor Sample

Testing the Patient Sample

DONOR RBC UNIT TESTING
According to Code of Federal Regulation and AABB
Standards
ABO TYPING
RH TYPING (including weak D)
CONFIRMATION OF RH NEGATIVE BLOOD
SEROLOGICAL TEST TO PREVENT BLOOD TRANSMITTED
DISEASES
ANTIBODY SCREENING FROM DONORS WHO HAVE
HISTORY OF BLOOD TRANSFUSION OR PREGNANCY
TESTING OF TRANSFUSION
RECIPIENT’S BLOOD SPECIMEN
Blood specimen acceptability
ABO and Rh typing
Antibody detection testing
Antibody identification
Comparison of current and previous results
DONOR RED CELL UNIT
SELECTION
SELECTION OF COMPONENTS OF ABO AND Rh TYPE THAT ARE
COMPATIBLE WITH THE RECIPIENT
USING THE SEGMENT FOR TESTING
DONOR AND RECIPIENT SAMPLES MUST BE STORED FOR 7
DAYS FOLLOWING TRANSFUSION
SAMPLES MUST BE STOPPERED, CAREFULLY LABELED AND
REFRIGERATED AT 1°C TO 6°C
MUST BE ADEQUATE IN VOLUME FOR RE-EVALUATION IF THE
RECIPIENT EXPERIENCE ANY ADVERSE REACTION
COMPATIBILITY TESTING
PROTOCOLS
TESTING DONOR SAMPLE
TESTING PATIENT SAMPLE
ABO GROUPING
Rh TYPING
ANTIBODY SCREENING
SELECTION OF APPROPRIATE DONOR UNITS
CROSSMATCHING
SUGGESTED ABO GROUP SELECTION
ORDER FOR TRANSFUSION OF RBCS
RECIPIENT 1ST CHOICE 2ND CHOICE 3RD CHOICE 4TH CHOICE
ABO GROUP

AB AB A B O

A A O

B B O

O O
SELECTION OF APPROPRIATE
DONOR UNIT
FIRST CHOICE SAME ABO AND Rh GROUP
IF NOT POSSIBLE PACKED RBC OF A UNIT , LACK OF ANTIGEN IN
WHICH THE RECIPIENT HAS AN ANTIBODY
Rh NEGATIVE BLOOD CAN BE GIVEN TO Rh POSITIVE
INDIVIDUAL ESPECIALLY IF THE UNIT NEAR EXPIRATION
Rh POSITIVE BLOOD SHOULD NOT BE GIVEN TO Rh NEGATIVE
FEMALE IN CHILD BEARING AGE
Rh NEGATIVE MALE AND FEMALE BEYOND MENOPAUSE CAN
RECEIVED Rh POSITIVE BLOOD AS LONG AS NO PREFORMED
ANTI-D IN THE SERA
CONT
IF UNEXPECTED ANTIBODY IS FOUND IN THE PATIENT SERUM
DURING ANTIBODY SCREENING IT IS BEST TO DO
CROSSMATCHING WITH INCUBATION AT 37°C AND AHG
IF THE ANTIBODIES REACTIVE BELOW 37°C ONLY NO NEED TO
PROVIDE ANTIGEN-NEGATIVE RBCS CAUSE THESE AB’S ARE
INCAPABLE OF CAUSING SIGNIFICANT RBC DESTRUCTION IN
VIVO
DONOR UNIT NEAR EXPIRY DATE SHOULD BE THE FIRST CHOICE
DONOR UNITS SHOULD BE EXAMINED VISUALLY FOR TURBIDITY,
COLOR, CLOTS, INCOMPLETE AND IMPROPER LABELINGOR
LEAKAGE
CROSSMATCHING
PART OF PRETRANSFUSION TESTING
PURPOSE
FINAL CHECK OF ABO COMPATIBILITY BETWEEN DONOR AND
PATIENT
MAY DETECT THE PRESENCE OF ANTIBODY IN THE PATIENT’S
SERUM THAT WILL REACT WITH ANTIGENS ON DONOR RBCS,
THAT WAS NOT DETECTED IN ANTIBODY SCREENING BECAUSE
THE CORRESPONDING ANTIGEN WAS LACKING FROM THE
SCREENING CELLS
SEROLOGIC CROSSMATCH TESTS
IMMEDIATE SPIN
TYPE AND SCREEN WITH IMMEDIATE SPIN= ABBREVIATED
CROSSMATCH
99.9 PERCENT EFFECTIVE IN PREVENTING OCCURRENCE OF
INCOMPATIBLE TRANSFUSION
DOES NOT DETECT ALL ABO INCOMPATIBILITIES SUCH US
AUTOANTI-I, PATIENT WITH HYPERIMMUNE ABO
ANTIBODIES AND WHEN INFANT SPECIMEN IS BEING
TESTED. CAN BE ENHANCES BY ADDING
ETHYLENEDIAMINETETRAACETIC TO THE TEST SYSTEM TO
ELIMINATE FALSE-POSITIVE REACTION
ANTIGLOBULIN CROSSMATCH
ADDITION OF AHG AFTER IMMEDIATE SPIN AND INCUBATION
AT 37°C
SEVERAL ENHANCEMENT MEDIA TO BOOST ANTIGEN-
ANTIBODY REACTIONS
LISS (LOW IONIC STRENGTH SOLUTION)
ALBUMIN
POLYETHYLENE GLYCOL
POLYBRENE
AUTOCONTROL CAN BE ALSO INCLUDED
INTERPRETATION OF RESULTS
INCOMPATIBLE: HEMOLYSIS AND AGGLUTINATION
COMPATIBLE: NO AGGLUTINATION , NO HEMOLYSIS
CAUSES OF POSITIVE RESULTS IN CROSSMATCHING(
INCOMPATIBLE)
1. INCORRECT ABO GROUPING OF THE PATIENT OR
DONOR
REPEAT IMMEDIATELY IF INCOMPATIBLE IN
IMMEDIATE SPIN
CAUSES OF POSITIVE
PRETRANSFUSION TESTS
Negative Antibody Screen, Incompatible Immediate-
Spin Crosshatch
Donor red cells are ABO incompatible
Donor red cells are polyagglutinable
Anti-A1 is the serum an A2 or A2B individual
Other Alloantibodies reactive at room temperature
(anti-M)
Rouleaux formation
Cold autoantibodies (anti-I)
Passively acquired anti-A or anti-B
CAUSES OF POSITIVE
PRETRANSFUSION TESTS
Negative antibody screen, Incompatible Antiglobulin
Crossmastch
Donor red cell have a positive DAT
Antibody reacts only with red cells having strong
expression of a particular antigen
Antibody to a low-incidence antigen on the donor
red cell
Passively acquired anti-A or anti-B
CAUSES OF POSITIVE
PRETRANSFUSION TESTS
Positive antibody screen, Compatible Crossmtaches
Autoanti-IH (autoanti-H) or anti-L ebH and nongroup
O units are selected
Antibodies dependent on reagent red cell diluent
Antibodies demonstrating dosage and donor red cell
are from heterozygotes (expressing single dose of
antigen)
Donor unit lacking of corresponding antigen
CAUSES OF POSITIVE
PRETRANSFUSION TESTS
Positive Antibody screen, incompatible crossmatch,
autocontrol negative
Alloantibody(ies)
Positive Antibody Screen, Incompatible Crossmatch,
Positive autocontrol, Negative DAT
Antibody to ingredient in enhancement media or
enhancement dependent antibody
Rouleaux formation
CAUSES OF POSITIVE
PRETRANSFUSION TESTS
Positive Antibody Screen, Incompatible Crossmatch,
Positive autocontrol, Positive DAT
Alloantibody causing either a delayed serologic or
haemolytic transfusion reaction
Passively acquired autoantibody (e.g. intravenous
immunoglobulin)
Cold-or warm-reactive autoantibody
Rouleaux formation
INCOMPATIBLE
CONTAMINANTS IN THE TEST SYSTEM
DIRTY GLASSWARE
BACTERIAL CONTAMINATION OF SAMPLES
CHEMICAL OR OTHER CONTAMINANTS IN THE
SALINE
FIBRIN CLOTS
PROVIDING COMPATIBLE BLOOD
PRODUCTS
Determined by the frequency of the antigen in the population
Helpful when determining the number of units that must be
antigen-typed to find a sufficient number to fill the crossmatching
request

Number of units/frequency of antigen

For example 2 units of RBC is needed and the patient has an anti-E
and the frequency of E-negative is 70%
The result is 2.8 or three units of blood would be needed to typed
for E antigen
In case of multiple specificities both frequency will be multiplied
PRETRANSFUSION TESTING IN
EMERGENCIES
ABO AND RH BLOOD GROUP SPECIFIC
LISS AND SHORTENED INCUBATION CAN BE EMPLOYED
IN EXTREME EMERGENCIES, TYPE O RH NEGATIVE PACKED RBC
CAN BE TRANSFUSED
ACCORDING TO CODE OF FEDREAL REGULATION
PRETRANSFUSION TESTING SHOULD BE COMPLETED AND ANY
INCOMPATIBILITY SHOULD BE REPORTED IMMEDIATELY TO THE
RECIPIENT PHYSICIAN AND THE BLOOD BANK MEDICAL
DIRECTOR
TRANSFUSION OF NON-GROUP
SPECIFIC BLOOD GROUP
WHEN BLOOD GROUP O IS TRANSFUSED TO A OR B INDIVIDUAL
, TESTING THE RECIPIENT SERUM FOR THE FOR THE PRESENCE
OF UNEXPECTED ANTI-A OR ANTI-B MUST BE PERFORMED
PRIOR TO GIVING ADDITIONAL RBC TRANSFUSION.
IF COMPATIBLE TO RECIPIENT OWN BLOOD GROUP , GIVE
SAME ABO SPECIFIC BLOOD GROUP
IF THERE IS INCOMPATIBILITY GROUP O PACKED RBC SHOULD
BE THE CHOICE OF BLOOD
COMPATIBILITY FOR PLASMA
PRODUCTS
COMPATIBILITY TESTING IS NOT REQUIRED
FOR TRANSFUSION OF PLASMA
PRODUCTS
IF LARGE VOLUME OF PLASMA PRODUCTS
DO MINOR CROSSMATCHING
JUST TO DETECT ABO INCOMPATIBILITY,
IMMEDIATE SPIN IS SUFFICIENT
INTRAUTERINE TRANSFUSIONS
BLOOD FOR INTRAUTERINE TRANSFUSION MUST BE
COMPATIBLE WITH MATERNAL ANTIBODIES CAPABLE OF
CROSSING THE PLACENTA
IF ABO AND Rh GROUP OF THE FETUS IS DETERMINED, GROUP
SPECIF BLOOD COULD BE GIVEN PROVIDED THAT THERE IS NO
FETO-MATERNAL ABO INCOMPATIBILITY
IF THE ABO AND Rh IS NOT KNOWN TYPE O RH NEGATIVE
SHOULD BE GIVEN, LACKIING WITH ANY ANTIGEN AGAINST
MOTHER SERUM
MOTHER SERUM SAMPLE IS USE FOR CROSSMATCHING
NEONATAL TRANSFUSION
YOUNGER THAN 4 MONTHS OF AGE, THE BLOOD UNIT FOR AN
EXCHANGE AND REGULAR TRANSFUSION MUST BE COMPATIBLE
WITH THE MATERNAL ANTIBODIES THAT HAVE ENTERED THE
INFANT CIRCULATION AND ARE REACTIVE AT 37°C OR AHG
BLOOD OF THE INFANTS ABO AND RH GROUP CAN BE USED
PROVIDED THAT IT IS NOT INVOLVED IN FETOMATERNAL
INCOMPATIBILITY
IF BLOOD GROUP OTHER THAN TYPE O , THE INFANT SERUM
MUST BE TESTED FOR THE PRESENCE OF ANTI-A OR ANTI-B
PRIOR TO TRANSFUSION OF ANOTHER UNIT
NO NEED TO REPEAT PRETRANSFUSION TESTING DURING ONE
HOSPITAL ADMISSION
BLOOD FOR TRANSFUSION MUST BE FRESH AND NO OLDER
THAN 7 DAYS
MASSIVE TRANSFUSIONS
ADMINISTRATION OF 8 TO 10 RBC UNITS TO AN ADULT PATIENT
IN LESS THAN 24 HOURS / 4 TO 5 RBC UNIT IN LESS THAN 1
HOUR
THE COMPATIBILITY TESTING IS SHORTENED OR ELIMINATED AS
LONG AS THE ATTENDING PHYSICIAN WILL SIGNED A WAIVER
FOR RELEASING UNTESTED UNIT FOR TRANSFUSION
IMPORTANT TO GIVE THE PATIENT THE CHANCE TO SURVIVE
AND THEN TREAT THE IMMUNE-MEDIATED ANEMIA INDUCED
BY MASSIVE TRANSFUSION
PREOPERATIVE AUTOLOGOUS
BLOOD
AUTOLOGOUS TRANSFUSION = REMOVAL OF BLOOD
OR COMPONENTS FROM A DONOR FOR THE DONOR’S
POSSIBLE USE AT A LATER TIME
ANTIBODY SCREENING AND CROSSMATCHING IS
OPTIONAL
THESE UNITS MUST BE LABELED ‘FOR AUTOLOGOUS
USE ONLY
PRETRANSFUSION TESTING
SCHEME
TEST SCHEME TEST PERFORMED ADVANTAGES LIMITATIONS
NO ORDER NONE NO SPECIMEN
COLLECTED, NO
TEST PERFORMED
HOLD CLOT NONE SPECIMEN NO TEST
COLLETED PERFORMED
TYPE AND HOLD ABO, RH SPECIMEN ANTIBODY
COLLECTED, SCREENING NOT
PATIENT’S ABO DONE
AND RH ARE
KNOWN
TYPE AND SCREEN ABO, RH AND MOST OF DOES NOT
ANTIBODY PRETRANSFUSION INCLUDE
SCREENING TESTING HAS CROSSMATCH
BEEN PERFORMED
TYPE AND COMPATIBILITY COMPATIBLE UNITS ARE
CROSSMATCH TESTING BLOOD CAN BE REMOVED FROM
LABELING AND REIDENTIFICATION
OF PATIENT PRIOR TRANSFUSION
LABEL THE COMPATIBLE BLOOD UNIT WITH THE PATIENT NAME
, FILE NUMBER OR ACCORDING TO THE SOP OF THE
LABORATORY
REIDENTIFICATION IS IMPERATIVE BEFORE BLOOD
TRANSFUSION
KEEPING THE RECORD OF THE PATIENT RECEIVING THE BLOOD
IS ALSO NECESSARY
FOLLOW THE SOP OF THE BLOOD BANK LABORATORIES
OVERVIEW OF THE
ROUTINE BLOOD
BANK
CHAPTER 11
BLOOD BANK LABORATORY
Facility involved in collection, storage, processing and
distribution of human blood and blood products for
transfusion
Operating under the constraints laws administered by
various governmental agencies in accordance with
standards established by AABB
Blood Bank Areas and Functions

Component Preparation and Storage

Separation of whole blood into packed RBCs,
plasma, platelets and cryoprecipitate
Storage of blood products at appropriate
temperatures
Apheresis procedures
Donor processing
Testing of donor units for ABO, Rh, antibody screen,
serologic test for syphilis, and transfusion
transmitted viruses
Blood Bank Areas and Functions
Product Labelling
RBCs and other components are labelled
Products are stored at proper temperatures
Main Laboratory
Patient sample are tested for ABO and Rh, Antibody
Screen, Crossmtach, DAT, Prenatal evaluation,
Postpartum evaluation, and Cord blood studies
Issue blood products
Blood Bank Areas and Functions
Reference Laboratory
Resolution of ABO and Rh discrepancies
Antibody identification
POSITIVE DAT
Warm autoantibodies
Cold autoantibodies
Transfusions reactions
Blood Bank with Collection
Facilities
Blood banks that collect their own units of blood
Separating to various components
Platelets- allowed to cool at room temperature
(20-24°C)
up to 24 hours after collection
FFP- allowed to cool at 1 to 10°C- must be
prepared within 8 hours after collection
Large, floor model temperature-controlled centrifuge
Apheresis machine
Blood Banks Without Collection
Facilities
Blood banks that depend on outside source for their
blood supplies usually receive their products in
component form
Automated cell washers- requiring washed RBCs
Sterile connecting devices
Produce sterile welds between two pieces
compatible tubing
APPLICATION OF STERILE
CONNECTING DEVICES
Adding fourth bag in a whole bag collection triple pack
Connecting additive solution to an RBC unit
Adding an in-line filter for leukocyte reduction
Pooling of blood components
Preparing aliquots for pediatric transfusion
Attaching processing solutions such as additive
solutions, glycerol for frozen RBCs and normal saline
to replace incompatible plasma
SUMMARY
MOST TRANSFUSION REACTIONS ARE CAUSED BY CLERICAL
ERRORS
COMPATIBILITY TESTING MUST BE PERFORMED PRIOR BLOOD
TRANSFUSIONS
ABO, RH, ANTIBODY SCREENING AND CROSSMTACHING
 SELECT APPROPRIATE DONOR FOR BLOOD TRANSFUSION,
MUST LACK OF ANTIGEN THAT WILL REACT TO PATIENT
ANTIBODIES
REFERENCES
HARMENING, D.M. MODERN BLOOD BANKING AND
TRANSFUSION PRACTICES