Manual Blood Transfusion Procedures

Full Blood Group

 Prepare a 5% suspension of cells to be tested in a 12 x 75mm test tube labelled with

the patient's name: Add 50µl of packed red cells to 1ml ID Diluent 2 and mix gently.

 Remove the aluminium foil strips from the pre-labelled BioRad DiaClon ABO/D +
reverse grouping card for each patient/control to be tested holding them in the upright
position.

 Pipette 50µl of ID-DiaCell A1 to microtube 5 (A1).

 Pipette 50µl of ID-DiaCell B to microtube 6 (B).

 Pipette 50µl of patient plasma to microtubes 5 and 6. Allow to incubate for 10

minutes at room temperature (18-25C).

 Pipette 10µl of the patient’s red cell suspension to microtubes 1-4 (A-B-D-ctl).

 Centrifuge the ID-cards for 10 minutes in the ID-Centrifuge (refer to SOP Operation &
Maintenance of DiaMed 12S Centrifuge).

 Read the card on the BioRad Banjo ID card reader (see section 3.5.2.5).

 The results are verified (see section 3.5.2.6) and then transferred via the interface
into Winpath. The result is recorded on the transfusion request form.

 Ensure the BioRad IH QC controls are tested and the correct reactions are obtained.

Antibody Screen

 Remove the aluminium foil strips from the pre-labelled LISS/Coombs card for each
patient/control to be tested card holding them in the upright position.

 Pipette 50µl of each ID-DiaCell I, II and III into the appropriate microtubes (marked
with the corresponding test cell).

 Pipette 25µl of patient plasma to each respective microtube.

  Incubate the cards for 15 minutes in the ID-Incubator (37C).

 Centrifuge the ID-cards for 10 minutes in the ID-Centrifuge.

 Read the card on the BioRad Banjo ID card reader (see section 3.5.2.5).

 The results are verified (see section 3.5.2.6) and then transferred via the interface
into Winpath. The result is recorded on the transfusion request form.

 Ensure the BioRad IH QC and NHSBT controls are tested and the correct reactions
are obtained.

Confirmation Group

 Prepare a 5% suspension of cells to be tested in a 12 x 75mm test tube labelled with

the patient's name: Add 50µl of packed red cells to 1ml ID Diluent 2 and mix gently.

 Remove the aluminium foil strips from the pre-labelled BioRad DiaClon ABD (VI-)
confirmation card for each patient/control to be tested, holding them in the upright
position.

 Pipette 10µl of the patient’s red cell suspension to microtubes (A-B-D).

 Centrifuge the ID-cards for 10 minutes in the ID-Centrifuge (refer to SOP Operation &
Maintenance of DiaMed 12S Centrifuge).

 Read the card on the BioRad Banjo ID card reader (see section 3.5.2.5).

 The results are verified (see section 3.5.2.6) and then transferred via the interface
into Winpath. The result is recorded on the transfusion request form.

 Ensure the BioRad IH QC controls are tested and the correct reactions are obtained.

Rh and K Phenotyping

 Prepare a 5% suspension of cells to be tested in a 12 x 75mm test tube labelled with

the patient's name: Add 50µl of packed red cells to 1ml ID Diluent 2 and mix gently.
  Remove the aluminium foil strips from the pre-labelled BioRad DiaClon Rh-
Subgroups + K card for each patient/control to be tested, holding them in the upright
position.

 Pipette 10µl of the patient’s red cell suspension to microtubes (A-B-D).

 Centrifuge the ID-cards for 10 minutes in the ID-Centrifuge (refer to SOP Operation &
Maintenance of DiaMed 12S Centrifuge).

 Read the card on the BioRad Banjo ID card reader (see section 3.5.2.5).

 The results are verified (see section 3.5.2.6) and then printed (via the Daily Journal in
IH-Com see Use of The BioRad IH-500 Analyser SOP). The results are entered
manually into Winpath. The printed results are attached to the transfusion request
form.

 Ensure the BioRad IH QC controls are tested and the correct reactions are obtained.

Cross-matching

Blood Selection

 Select matching ABO & Rh(D) donor units from the blood bank refrigerator. Where
possible ABO Rh(D) units will be ordered to meet known requirements, however if
ABO & Rh(D) group specific blood is not present in stock compatible selections can
be made according to the donor unit selection table in Appendix 2 of the Use of the
BioRad IH-500 Analyser SOP.

 Blood for women under the age of 60 years must be negative for the K antigen.

 RhD positive blood should not be issued to RhD negative women under the age of 60
years. Guidelines should be followed but in an emergency/urgent situation the
prescribing clinician may decide to give RhD positive blood. Advice can be obtained
from the Consultant Haematologist.

 For RhD negative males and women >60 years with no known anti-D O RhD positive
blood can be considered for transfusion in an emergency situation.

 For patients with known clinically significant red cell alloantibodies it may be
necessary to specifically order antigen negative blood (refer to the donor unit
selection table for patients with red cell alloantibodies in Appendix 3 of the Use of the
BioRad IH-500 Analyser SOP).
 For patients with unidentifiable red cell alloantibodies samples must be referred to
NHSBT/SNBTS for testing, antibody identification and provision of compatible blood
(refer to SOP Referral of Samples to NHSBT). In an emergency situation it may be
possible to issue blood to these patients provided the serological cross-match is
negative (refer to SOP Exceptional Release of Blood Components).

 For patients with autoimmune haemolytic anaemia, sickle cell anaemia or recipients
of allogenic haemopoietic stem cell grafts it may be advisable to refer samples to the
NHSBT/SNBTS for cross-matching and/or to provide Rh and K phenotypically
matched compatible blood. If in doubt, discuss with the Consultant Haematologist
(refer to SOP Procedure for Contacting Consultant Haematologist).

 It may be necessary to return units from the issue fridge that have been allocated for
remote issue, refer to SOP Moving Blood Products in Blood360.

 Ensure units to be cross-matched are available.

 Check all blood units for any signs of deterioration i.e. evidence of haemolysis,
discoloration, turbidity, clots etc. prior to issue. Gently apply pressure to the units to
check for any leakage.

 Units showing signs of deterioration or leakage must not be issued. De-select these
units and place in the quarantine section of the fridge. Refer to SOP Recall of Blood
Components.

Cross-match Procedure

 Ensure all reagents and cards are in date and validated.

 Diluent should be allowed to reach room temperature before use.

For the first donor unit to be tested, proceed as follows:

 Clearly label two 12 x 75mm test tubes with (at least) the last 7 digits of the donation
number of the donor unit.

 Using a “transclip”, or similar line sealing device, seal one end of the line from the
donor pack approximately 3-4 cm from the free end to ensure packed cells can be
accessed.

 Cut the line just beneath the clip leaving the clip attached to the line on the bag.
  Dispense approximately 0.5ml of packed cells from the line to one of the
correspondingly labelled 12x75 tubes. There is no requirement to wash the cells. Mix
by swirling gently.

 To the paired tube, add 1ml ID Diluent 2.

 Repeat for each of the remaining units in turn.

 Prepare a 0.8% (±0.3%) suspension of donor cells: for each of the paired tubes add
10µl of packed red cells from the first tube to the ID Diluent 2 in the second tube and
mix gently.

 Prepare a 0.8% (±0.3%) suspension of patient cells in a labelled 12 x 75mm test

tube: Add 10µl of packed red cells to 1ml ID Diluent 2 and mix gently.

 Remove the aluminium foil strips from each pre-labelled LISS/Coombs card holding
them in the upright position.

 Pipette 50µl of the donor red cell suspension to the appropriate microtube.

 Pipette 25µl of patient plasma to each microtube labelled as donor numbers.

 Incubate the cards for 15 minutes at 37C in the ID-Incubator.

 Centrifuge the ID-cards for 10 minutes in the ID-Centrifuge.

 Read the card on the BioRad Banjo ID card reader (see section 3.5.2.5).

 The results are verified (see section 3.5.2.6) and then printed (via the Daily Journal in
IH-Com see Use of The BioRad IH-500 Analyser SOP). The results are entered
manually into Winpath. The printed results are attached to the transfusion request
form.

Note: If an auto control is required the test cannot be requested via IH-Com and a
LISS/Coombs card must be labelled with the patient’s name, sample and donation numbers
manually along with the auto and positive control. Pipette 50µl of the patient’s red cell
suspension to the labelled auto control microtube and add 25µl of patient plasma. Pipette
50µl of BioRad ID-Diacell Screening Cell I to the positive control microtube and add 25µl of
Weak anti-D. Ensure the weak anti-D gives the appropriate reaction. Record results
on the Transfusion Request Form or the Manual Cross-Match Worksheet in
Appendix 2.

Antibody Investigation
IAT Panels

Antibody investigation panels should be performed in conjunction with a Direct Antiglobulin

Test.

 Prepare a 0.8% (±0.3%) suspension of patient cells in a labelled 12 x 75mm test

tube: add 10µl of packed red cells to 1ml ID Diluent 2 and mix gently.

 Label two BioRad LISS/Coombs cards with the patient’s name and specimen number
and identify a micro tube for each panel cell (1 to 11).

 Remove the aluminium foil strips from each card holding them in the upright position.

 Carefully dispense 50µl of the IAT panel cells to the appropriate LISS/Coombs
microtube.

 Pipette 25µl of patient plasma to each microtube.

 Incubate the cards for 15 minutes in the ID-Incubator (37C).

 Centrifuge the ID-cards for 10 minutes in the ID-Centrifuge.

 Read the card on the BioRad Banjo ID card reader (see section 3.5.2.5).

 Using Daily Journal in IH-Com print the results and attach them to the transfusion
request form.

 Record the reactions on a copy of the antigen table supplied with the panel cells
labelled with the patient’s details. Enter results onto Winpath. A second BMS should
also check the results to minimise transcription errors.

Note: If an auto control is required the test cannot be requested via IH-Com and a
LISS/Coombs card must be labelled with the patient’s name, and sample number manually.
Label the wells including the auto control. Pipette 50µl of the patient’s red cell suspension to
the labelled auto control microtube and add 25µl of patient plasma.

Enzyme Panels

 Prepare a 0.8% (±0.3%) suspension of patient cells in a labelled 12 x 75mm test

tube: add 10µl of packed red cells to 1ml ID Diluent 2 and mix gently.
  Label two BioRad NaCl/enzyme cards with the patient’s name and specimen number
and a micro tube for each panel cell (1 to 11).

 Remove the aluminium foil strips from each card holding them in the upright position.

 Carefully dispense 50µl of the enzyme panel cells to the appropriate NaCl/enzyme
microtube.

 Pipette 25µl of patient plasma to each microtube.

 Incubate the cards for 15 minutes in the ID-Incubator (37C).

 Centrifuge the ID-cards for 10 minutes in the ID-Centrifuge.

 Read the card on the BioRad Banjo ID card reader (see section 3.5.2.5).

 Using Daily Journal in IH-Com print the results and attach them to the transfusion
request form.

 Record the reactions on copy of the antigen table supplied with the panel cells
labelled with the patient’s details. Enter the results onto Winpath. A second BMS
should also check the results to minimise transcription errors.

Direct Antiglobulin Test (DAT)

 Include a negative control, either BioRad IH QC sample 1 or 2.

 Prepare a 0.8% (±0.3%) suspension of cells to be tested in a 12 x 75mm test tube

labelled with the patient's name: Add 10µl of packed red cells to 1ml ID Diluent 2 and
mix gently.

 Label a BioRad LISS/Coombs card with the name and specimen number on one
microtube position for each patient/control to be tested.

 Remove the aluminium foil strips from each card holding them in the upright position.

 Pipette 50µl of the red cell suspension to the appropriate microtube.

 Centrifuge the ID-cards for 10 minutes in the ID-Centrifuge.

 Read the card on the BioRad Banjo ID card reader (see section 3.5.2.5).
 Using Daily Journal in IH-Com print the results and attach to the transfusion request
form.

 Enter the results onto Winpath.