CROSSMATCH TECHNIQUES

An Educational Supplement prepared by ALQEP February 2012

Introduction
Transfusion medicine standards require the performance of a crossmatch to detect incompatibility between the
recipient plasma and donor red cells before transfusion of red cell components. A number of options can be used to
accomplish this objective, each with prerequisite requirements and limitations. Traditionally, a serologic crossmatch
using antiglobulin technique was performed prior to all red cell transfusions. Changes in standards now allow
abbreviation of the serologic crossmatch or the use of an electronic (computer) crossmatch under defined
conditions.

Serologic Crossmatch
A serologic crossmatch is an in vitro test performed to determine the compatibility of the recipient plasma and donor
red cells.
When testing of the recipient plasma indicates the presence of a clinically significant antibody, or the recipient has a
history of a previously detected clinically significant antibody, the serologic crossmatch must include incubation at
37C and be performed by an antiglobulin or comparable technique. These techniques may also be used for
crossmatches of recipients without antibodies if the laboratory chooses this process.
The antiglobulin technique may be a saline indirect antiglobulin test (SIAT) or the technique may include the addition
of a potentiating agent such as 22% bovine serum albumin (AIAT), low ionic strength solution LISS (LIAT), or
polyethylene glycol PEG (PIAT). In a 2009 ALQEP participant survey, 17 % of respondents routinely performed an
antiglobulin crossmatch (1% - AIAT; 3% - LIAT; 11% - PIAT; 2% - SIAT) for all compatibility testing, regardless of
recipient antibody status. Preliminary data from a 2011 participant survey indicates that this number has decreased
to 12% of participants (1% - LIAT, 11% - PIAT).
Techniques comparable to an antiglobulin crossmatch include gel and solid phase technologies (manual or
automated). In 2009, 19% of respondents performed routine crossmatches by gel technology (18% manual, 1%
automated) and 2% utilized solid phase technology (1% manual, 1% automated). In the 2011 preliminary data,
19% of respondents utilized a routine gel crossmatch (16% manual, 3% automated). No respondents indicated the
use of solid phase technology for crossmatch in 2011.
When the recipients antibody screen is negative and there is no history of a previously detected clinically significant
antibody, standards allow the use of an immediate spin crossmatch. The purpose of the immediate spin crossmatch
is to detect ABO incompatibility between the recipients plasma and the donor red cells. In 2009, 40% of ALQEP
respondents routinely utilized the immediate spin crossmatch. 2011 preliminary data shows a similar rate of 39%.

Copyright 2012 College of Physicians and Surgeons of Alberta

CROSSMATCH TECHNIQUES
Laboratories must consider the limitations of a serologic crossmatch and the relationship of the serologic crossmatch
to the other components of pre-transfusion testing:
In April 2011, Ortho Clinical Diagnostics (OCD) issued a Customer Information Letter describing the
limitations of the MTSTM Anti-IgG Card for crossmatch compatibility testing. OCD state that this card is
intended to be used for antiglobulin crossmatches to detect IgG antibodies only and does not meet the
requirements of detecting ABO incompatibility due to IgM antibodies. Therefore, when an MTSTM Anti-IgG
Card is used for crossmatch purposes, an additional method must be used to confirm ABO compatibility
between the recipient plasma and donor red cells. This may be accomplished by the performance of:
o tube immediate spin crossmatch
o MTSTM Buffered Gel Card immediate spin crossmatch
o electronic crossmatch (when all eligibility requirements for electronic crossmatch are met)
Solid phase testing is also designed for the detection of IgG antibodies and therefore, when solid phase
technology is used for crossmatching, an additional technique, such as tube immediate spin or electronic
crossmatch, must be performed to verify ABO compatibility.
As standards require that ABO compatibility between the recipient and donor must be confirmed prior to
transfusion, it is important to identify cases where the recipients ABO antibodies are not demonstrable in the
plasma sample. In these cases, a serologic crossmatch will not adequately ensure ABO compatibility and the
laboratory must perform serologic confirmation of the donor ABO type to demonstrate compatibility with the
recipient.
In the presence of a positive antibody screen, an immediate spin crossmatch will often appear compatible. This
technique is not valid in this situation and should not be reported as compatible. The positive antibody screen
must be investigated and if a clinically significant antibody is identified, an antiglobulin or equivalent crossmatch
must be performed prior to transfusion of the donor red cells.
In the presence of a clinically significant antibody or the history of a previously detected clinically significant
antibody that is no longer reactive, an antiglobulin or equivalent crossmatch must be performed. Standards
indicate that the red cells selected for crossmatch should lack the corresponding antigen(s). The serologic
crossmatch may appear compatible with donor cells that are positive for the corresponding antigen(s) due to
variations in antigen expression (e.g., heterozygous cells).

Electronic Crossmatch
Laboratories may choose to perform an electronic crossmatch a computerized process to verify ABO compatibility
between the recipient plasma and donor red cells. This process cannot be used for recipients with clinically
significant antibodies or a history of a previously detected clinically significant antibody. In 2009, 22% of
respondents routinely used an electronic crossmatch when applicable. 2011 preliminary data shows an increase of
this rate to 30%.
The electronic crossmatch process must be validated to prevent the release of ABO incompatible blood
components. Requirements for the process are:
The recipients ABO blood group shall be determined twice. This includes testing of a current sample plus one
of the following:
o a second independent testing of the same sample.
Page 2 of 4

Alberta Laboratory Quality Enhancement Program

Copyright 2012 College of Physicians and Surgeons of Alberta

CPSA February 2012

CROSSMATCH TECHNIQUES
o
o

testing of a second correctly identified and labelled sample.

verification of previous ABO typing of the recipient on file.

The ABO type of the donor red cells shall be confirmed by the laboratory.
The computer system is designed to alert the laboratory to:
o discrepancies between the recipient current blood group results and previous blood group results.
o discrepancies between donor unit labelling and ABO confirmatory test interpretation.
o discrepancies between recipient and donor ABO groups.

Emergency Requirements for Transfusion

Transfusion Medicine Standards require that laboratories have a policy for abbreviated serologic testing when a
recipient receives a massive transfusion. The Transfusion Service Medical Director is responsible for establishing
this policy.
The laboratory also must have a policy for the release of blood components prior to the completion of
pretransfusion testing. In this situation:
the ABO group of the donor red cells must be serologically confirmed by the laboratory.
group O red cells and AB plasma shall be issued if the ABO and Rh group of the recipient has not been
determined on a current sample. Group O Rh negative red cells should be issued for women of
childbearing age and children.
the requesting physician must document that the clinical situation justifies the transfusion and obtain
informed consent from the recipient when possible.
the label attached to the component must indicate testing is incomplete and the information must be
documented in the recipients medical record.
pretransfusion testing shall be completed as soon as possible, and the attending physician and TS Medical
Director shall be informed if red cells are subsequently found to be incompatible.

Page 3 of 4

Alberta Laboratory Quality Enhancement Program

Copyright 2012 College of Physicians and Surgeons of Alberta

CPSA February 2012

CROSSMATCH TECHNIQUES
References:
1. Ortho Clinical Diagnostics. Customer information letter: The use of the Anti-Human Globulin Anti-IgG
(Rabbit) MTSTM anti-IgG Card and the MTSTM Buffered Gel Card for Crossmatch Compatibility Testing.
CL11-010a, April 27, 2011.
2.

Reagents / methods in use by ALQEP transfusion medicine participants. Edmonton AB: Alberta
Laboratory Quality Enhancement Program, 2009.

3. Roback JD, ed. Technical manual. 16th ed. Bethesda MD: AABB, 2008.
4. Standards for blood banks and transfusion services 27th ed. Bethesda MD: AABB, 2011.
5. Standards for hospital transfusion services Version 3. Ottawa ON: Canadian Society for Transfusion
Medicine, 2011.
6. Z902-10 Blood and blood components. Mississauga ON: Canadian Standards Association, 2010.

Page 4 of 4

Alberta Laboratory Quality Enhancement Program

Copyright 2012 College of Physicians and Surgeons of Alberta

CPSA February 2012