Ortho BioVue-Work Station System Handbook

A1 ORTHO BioVue® Cassette Description Foil Strip

6 columns

Product Name

Bar Code 20 Digits 6 digits

Expiry date

2 digits

Cassette type

6 digits

Sequential N°

6 digits

Lot N°

Bar Code 20 Digits

Lot N° & Expiration Date

Reagent Identification

Patient Identification

Results

Reaction chamber Air gap

Reaction Glass beads

SYSTEM PRESENTATION

A Page 1

A2 ORTHO™ Workstation Description

Centrifuge Cycle Speed

Phase 1 – 793 rpm ± 10 rpm Phase 2 – 1509 rpm ± 10 rpm

Speed Indicator

4 digits

Timing Indicator

4 digits

Cycle Timing

Phase 1 – two minutes ± 10 sec. (including ramp-up time) Phase 2 – three minutes ± 10 sec. (including ramp-up time)

Centrifuge Cassette Capacity

10 cassettes

Incubator

A Page 2

Incubation Temperature

37°C ± 2°C

Incubator Cassette Capacity

20 cassettes (two sections of 10)

SYSTEM PRESENTATION

A3 ORTHO BioVue® Pipette Description

To change the program, press M When the desired program appears, press E Choose the number of dispenses To confirm, press again E

PROGRAM

NUMBER

TAKEN

DELIVERED

P1

1 g 6x

40 g 240 µl

40 µl

P2

1 g 24x

10 g 240 µl

10µl

P3

1 g 6X

50 g 300 µl

50 µl

P4

1 g 6X

60 g 360 µl

60 µl

190µl + 10 µl

200 µl

P5

IMPORTANT: In the specific procedures sheets, a symbol like P1 refers to the electronic pipette program to use.

SYSTEM PRESENTATION

A Page 3

A4 Procedure to be Applied to ORTHO BioVue® Cassettes 1. Visual control of the ORTHO BioVue® cassettes + Do not use reagents beyond their labeled expiration date. + Do not use cassettes that appear damaged (i.e., break in foil seal or break, crack or bubble in the column) or exhibit drying (i.e., liquid level is at or below the top of the glass beads) or exhibit discoloration (due to bacterial contamination which can cause false reactions).

2. Sample identification and preparation

5. Sample pipetting

3. Cassettes identification and preparation

6. Incubation – Centrifugation

4. Reagent pipetting

7. Reading

A Page 4

SYSTEM PRESENTATION

A5 Sample Preparation 1. Sample identification 2. Sample centrifugation 5’ 900 – 1000 RPM

3. Sample preparation

RBC and Serum

4. RBC suspension preparation – A6 or A7 Maximum storage time at 2 g 8°C Anticoagulant

EDTA

Oxalate

Heparin

Citrate

No anticoagulant

Days

7

2

2

7

7

Donor RBC Preparation

1. Optional: Wash red blood cells one time in isotonic saline. 2. Donor RBC suspension preparation – A6 or A7

SYSTEM PRESENTATION

A Page 5

A6 Preparation of 4% Suspensions 1. ORTHO BioVue® electronic pipette – Program P5 190 µl NaCl + 10 µl packed red blood cells = 200 µl 4% red blood cells suspension

2. Manual preparation 1 ml NaCl + 50 µl packed red blood cells

A7 Preparation of 0.8% Suspensions 1. In Saline: 1 ml Saline + 10 µl packed red blood cells

2. In 0.8% ORTHO® Red Cell Diluent – for 0.8% Cross-Match: 1 ml 0.8% ORTHO® Red Cell Diluent + 10 µl packed donor red blood cells Maximum storage time at 2 g 8°C: 24 hours

A Page 6

SYSTEM PRESENTATION

B1 ABO/D Typing 1. Cassettes BioVue® ABO-Rh (A, B, AB, D, CDE, Control) or BioVue® ABODD (A, B, AB, D, D, Control) or BioVue® ABDD/K (A, B, D, D, Kell, Control) or BioVue® ABD Confirmation (A, B, D, A, B, D) Important: cassettes to be used with the liner!

2. Sample 4% patient red blood cells suspensions in saline (A6) or 0.8% patient red blood cells suspensions in saline (A7)

3. 4% Procedure + Identify the cassette + Add 10 µl 4% patient red blood cells suspensions - P2 + 5 minutes centrifugation + Reading (both sides)

4. 0.8% Procedure + Identify the cassette + Add 50 µl 0.8% patient red blood cells suspensions – P3 + 5 minutes centrifugation + Reading (both sides)

SpeCific Procedures - TYPING/SCREENING/CROSS-MATCH

B Page 7

B2 ABO Reverse Typing 1. Cassettes BioVue® Reverse Diluent (6 x Reverse)

2. Reagents Affirmagen™ 4 (A1, A2, B & O cells) or 0.8% Affirmagen™ 4 (A1, A2, B & O cells) or Affirmagen™ 2 (A1 & B cells) or 0.8% Affirmagen™ 2 (A1 & B cells)

3. Sample Patient Serum/Plasma

4. 4% Procedure + Identify the cassette + Add 10 µl Affirmagen™ 2 (A1, A2, B & O cells) - P2 + Add 40 µl patient serum/plasma - P1 + 5 minutes centrifugation + Reading (both sides)

5. 0.8% Procedure + Identify the cassette + Add 50 µl 0.8% Affirmagen™ 2 (A1, A2, B & O cells) - P3 + Add 40 µl patient serum/plasma - P1 + 5 minutes centrifugation + Reading (both sides)

B Page 8

SpeCific Procedures - TYPING/SCREENING/CROSS-MATCH

B3 ABO/D Typing & ABO Reverse 1. Cassettes BioVue® ABO-Rh D Combo (A, B, D, Control, Reverse, Reverse) Important: cassettes to be used with the liner!

2. Reagents Affirmagen™ 2 (A1 & B cells) or 0.8% Affirmagen™ 2 (A1 & B cells)

3. Sample 4% patient or donor red blood cells suspensions (A6) or 0.8% patient or donor red blood cells suspensions (A7) and Patient Serum/Plasma

4. 4% Procedure + Identify the cassette + Add 10 µl Affirmagen™ 2 (A1 & B cells) in the reverse columns- P2 + Add 40 µl patient serum/plasma in the reverse columns - P1 + Add 10 µl 4% patient red blood cells suspensions in the A, B, D & Control columns - P2 + 5 minutes centrifugation + Reading (both sides)

5. 0.8% Procedure + Identify the cassette + Add 50 µl 0.8% Affirmagen™ 2 (A1 & B cells) in the reverse columns - P3 + Add 40 µl patient serum/plasma in the reverse columns P1 + Add 50 µl 0.8% patient red blood cells suspensions in the A, B, D & Control columns – P3 + 5 minutes centrifugation + Reading (both sides)

SpeCific Procedures - TYPING/SCREENING/CROSS-MATCH

B Page 9

B4 ABO/D + Direct Coombs Newborn 1. Cassettes BioVue® Newborn (A, B, AB, D, Control, AHG anti-IgG) Important: cassettes to be used with the liner!

2. Sample 4% patient red blood cells suspensions in saline (A6) or 0.8% patient red blood cells suspensions in saline (A7)

3. 4% Procedure + Identify the cassette + Add 10 µl 4% patient red blood cells suspensions - P2 + 5 minutes centrifugation + Reading (both sides)

4. 0.8% Procedure + Identify the cassette + Add 50 µl 0.8% patient red blood cells suspensions – P3 + 5 minutes centrifugation + Reading (both sides)

B Page 10

SpeCific Procedures - TYPING/SCREENING/CROSS-MATCH

B5 RH/K Phenotyping 1. Cassettes BioVue® Rh/K (C, E, c, e, K, Control) BioVue® Rh-hr (D, C, E, c, e, Control) BioVue® Kell ( 6 x K) BioVue® Kell/Control (K, Control, K, Control, K, Control) Important: cassettes to be used with the liner!

2. Sample 4% patient red blood cells suspensions in saline (A6) or 0.8% patient red blood cells suspensions in saline (A7)

3. 4% Procedure + Identify the cassette + Add 10 µl 4% patient red blood cells suspensions - P2 + 5 minutes centrifugation + Reading (both sides)

4. 0.8% Procedure + Identify the cassette + Add 50 µl 0.8% patient red blood cells suspensions – P3 + 5 minutes centrifugation + Reading (both sides)

SpeCific Procedures - TYPING/SCREENING/CROSS-MATCH

B Page 11

B6 Direct Coombs 1. Cassettes BioVue® Anti-Human Globulin Polyspecific or BioVue® Anti-Human Globulin anti-IgG

2. Sample 4% patient red blood cells suspensions in saline (A6)

3. Procedure + Identify the cassette + Add 10 µl 4% patient red blood cells suspensions - P2 + 5 minutes centrifugation + Reading (both sides)

B Page 12

SpeCific Procedures - TYPING/SCREENING/CROSS-MATCH

B7 Direct Coombs Specific 1. Cassettes BioVue® DAT/IDAT (AHG anti-IgG, anti-C3b & -C3d, Control)

2. Sample 4% patient red blood cells suspensions in saline (A6)

3. Procedure + Identify the cassette + Add 10 µl 4% patient red blood cells suspensions - P2 + 5 minutes centrifugation + Reading (both sides)

SpeCific Procedures - TYPING/SCREENING/CROSS-MATCH

B Page 13

B8 4% Antibody Screening Indirect Coombs Technique 1. Cassettes BioVue® Anti-Human Globulin Polyspecific or BioVue® Anti-Human Globulin anti-IgG

2. Reagents Selectogen® (2 cells) or Surgiscreen® (3 cells) and Ortho® BLISS

3. Sample Patient Serum/Plasma

4. Procedure + Identify the cassette + Add 50 µl Ortho BLISS – P3 + Add 10 µl 4% Test Red Blood Cells - P2 + Add 40 µl patient serum/plasma – P1 + 10 – 15 minutes incubation (Max. 30 minutes) + 5 minutes centrifugation + Reading (both sides)

B Page 14

SpeCific Procedures - TYPING/SCREENING/CROSS-MATCH

B9 0.8% Antibody Screening Indirect Coombs Technique 1. Cassettes BioVue® Anti-Human Globulin Polyspecific or BioVue® Anti-Human Globulin anti-IgG

2. Reagents 0.8% Selectogen® (2 cells) or 0.8% Surgiscreen® (3 cells)

3. Sample Patient Serum/Plasma

4. Procedure + Identify the cassette + Add 50 µl 0.8% Test Red Blood Cells – P3 + Add 40 µl patient serum/plasma – P1 + 10 – 15 minutes incubation (Max. 30 minutes) + 5 minutes centrifugation + Reading (both sides)

SpeCific Procedures - TYPING/SCREENING/CROSS-MATCH

B Page 15

B10 4% Antibody Screening IAT & Enzyme Technique 1. Cassettes BioVue® Anti-Human Globulin Polyspecific/Neutral

2. Reagents BioVue® Screen Ficin (3 untreated & 3 ficin-treated cells) and Ortho® BLISS

3. Sample Patient Serum/Plasma

4. Procedure + Identify the cassette + Add 50 µl Ortho BLISS in the 3 Polyspecific Columns – P3 + Add 10 µl 4% untreated Test Red Blood Cells in the 3 Polyspecific Columns - P2 + Add 10 µl 4% ficin-treated Test Red Blood Cells in the 3 neutral Columns - P2 + Add 40 µl patient serum/plasma in the 6 columns – P1 + 10 – 15 minutes incubation (Max. 30 minutes) + 5 minutes centrifugation + Reading (both sides)

B Page 16

SpeCific Procedures - TYPING/SCREENING/CROSS-MATCH

B11 0.8% Antibody Screening IAT & Enzyme Technique 1. Cassettes BioVue® Anti-Human Globulin Polyspecific/Neutral

2. Reagents 0.8% BioVue® Screen Ficin (3 untreated & 3 ficin-treated cells)

3. Sample Patient Serum/Plasma

4. Procedure + Identify the cassette + Add 50 µl 0.8% untreated Test Red Blood Cells in the 3 Polyspecific Columns – P3 + Add 50 µl 0.8% ficin-treated Test Red Blood Cells in the 3 neutral Columns – P3 + Add 40 µl patient serum/plasma in the 6 columns – P1 + 10 – 15 minutes incubation (Max. 30 minutes) + 5 minutes centrifugation + Reading (both sides)

SpeCific Procedures - TYPING/SCREENING/CROSS-MATCH

B Page 17

B12 Cross-Match 4% Indirect Coombs Technique 1. Cassettes BioVue® Anti-Human Globulin Polyspecific or BioVue® Anti-Human Globulin anti-IgG

2. Reagents Ortho® BLISS

3. Sample Patient Serum/Plasma 4% donor red blood cells suspensions in Saline (A6)

4. Procedure + Identify the cassette + Add 50 µl Ortho BLISS in the 3 Polyspecific Columns – P3 + Add 10 µl 4% donor red blood cells - P2 + Add 40 µl patient serum/plasma – P1 + 10 – 15 minutes incubation (Max. 30 minutes) + 5 minutes centrifugation + Reading (both sides)

B Page 18

SpeCific Procedures - TYPING/SCREENING/CROSS-MATCH

B13 Cross-Match 0.8% Indirect Coombs Technique 1. Cassettes BioVue® Anti-Human Globulin Polyspecific or BioVue® Anti-Human Globulin anti-IgG

2. Reagent 0.8% Ortho® Red Cell Diluent

3. Sample Patient Serum/Plasma 0.8% donor red blood cells suspensions in 0.8% Ortho® Red Cell Diluent (A7)

4. Procedure + Identify the cassette + Add 50 µl 0.8% donor red blood cells – P3 + Add 40 µl patient serum/plasma – P1 + 10 – 15 minutes incubation (Max. 30 minutes) + 5 minutes centrifugation + Reading (both sides)

SpeCific Procedures - TYPING/SCREENING/CROSS-MATCH

B Page 19