JOURNAL TRANSCRIPT
ORTHO BioVue® System Handbook
Table of Contents
A
SYSTEM PRESENTATION
A1
BioVue® Cassette Description
A2
BioVue® System Description
A3
BioVue® Electronic Pipette Description
A4
Procedure to be applied to BioVue® Cassettes
A5
Sample Preparation
A6
Preparation of 4% Suspensions
A7
Preparation of 0.8% Suspensions
B
SPECIFIC PROCEDURES - TYPING/SCREENING/CROSS-MATCH
B1
ABO/D Typing
B2
ABO reverse typing
B3
ABO/D Typing & ABO reverse typing
B4
ABO/D + Direct Coombs – Newborn
B5
Rh/K Phenotyping
B6
Direct Coombs
B7
Direct Coombs specific
B8
4% Antibody Screening – Indirect Coombs Technique
B9
0.8% Antibody Screening – Indirect Coombs Technique
B10
4% Antibody Screening – IAT & Enzyme Technique
B11
0.8% Antibody Screening – IAT & Enzyme Technique
B12
4% Cross-Match - Indirect Coombs Technique
B13
0.8% Cross-Match - Indirect Coombs Technique
B14
4% Cross-Match - Enzyme Technique
B15
4% Antibody Identification - Indirect Coombs Technique
B16
0.8% Antibody Identification - Indirect Coombs Technique
B17
4% Antibody Identification - Enzyme Technique
B18
0.8% Antibody Identification - Enzyme Technique
B19
Extended Phenotyping – Anti-Fya, anti- Fyb, Anti-S, anti-s, anti-D (IAT)
B20 Extended Phenotyping – Anti-Jka, anti-Jkb, Anti-K, anti-D (DVI), anti-P1, anti-Lea, anti-Leb
B21
Extended Phenotyping – Anti-M, anti-N
C
ORTHO BIOVUE® RESULTS INTERPRETATION
C1
Grading Chart
C2-C3 Difficulties Solving/Troubleshooting
IMPORTANT: All the information included in this hand-book are extracted from the BioVue® cassettes « Instruction for Use » which are the only authorized source for information.
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® BioVue® BioVue® BioVue®
ABO-Rh (A, B, AB, D, CDE, Control) or ABODD (A, B, AB, D, D, Control) or ABDD/K (A, B, D, D, Kell, Control) or 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 + + + + +
B Page 8
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)
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 + + + +
B Page 10
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
B5 RH/K Phenotyping 1. Cassettes
BioVue® BioVue® BioVue® BioVue®
Rh/K (C, E, c, e, K, Control) Rh-hr (D, C, E, c, e, Control) Kell ( 6 x K) 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 + + + +
B Page 12
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
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 + + + + + + +
B Page 14
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)
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 + + + + + + +
B Page 18
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)
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
B14 Cross-Match 4% Enzyme Technique 1. Cassettes BioVue® Neutral
2. Reagents Enzyme Solution
3. Sample Patient Serum/Plasma 4% donor red blood cells suspensions in Saline (A6)
4. Procedure + + + + + + +
B
Identify the cassette Add 40 µl Enzyme – P1 Add 10 µl 4% donor red blood cells - P2 Add 40 µl patient serum/plasma – P1 10 – 15 minutes incubation 5 minutes centrifugation Reading (both sides)
SpeCific Procedures - TYPING/SCREENING/CROSS-MATCH
Page 20
B15 4% Antibody Identification Indirect Coombs Technique 1. Cassettes BioVue® Anti-Human Globulin Polyspecific or BioVue® Anti-Human Globulin anti-IgG
2. Reagents Resolve® Panel A (11 cells), Resolve® Panel B (11 cells) or Resolve® panel (R) C (11 cells untreated) 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)
SpeCific Procedures - TYPING/SCREENING/CROSS-MATCH
B Page 21
B16 0.8% Antibody Identification Indirect Coombs Technique 1. Cassettes BioVue® Anti-Human Globulin Polyspecific or BioVue® Anti-Human Globulin anti-IgG
2. Reagents 0.8% Resolve® Panel A (11 cells), 0.8% Resolve® Panel B (11 cells) or 0.8% Resolve® Panel (R) C (11 cells untreated)
3. Sample Patient Serum/Plasma
4. Procedure + + + + + +
B Page 22
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
B17 4% Antibody Identification Enzyme Technique 1. Cassettes BioVue® Neutral
2. Reagents Resolve® Panel C (the 11 ficin-treated cells)
3. Sample Patient Serum/Plasma
4. Procedure + + + + + +
Identify the cassette Add 10 µl ficin-treated 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)
SpeCific Procedures - TYPING/SCREENING/CROSS-MATCH
B Page 23
B18 0.8% Antibody Identification Enzyme Technique 1. Cassettes BioVue® Neutral
2. Reagents 0.8% Resolve® Panel C (the 11 ficin-treated cells)
3. Sample Patient Serum/Plasma
4. Procedure + + + + + +
B
Identify the cassette Add 50 µl ficin-treated 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
Page 24
B19 Extended Phenotyping – Anti-Fya, anti- Fyb, Anti-S, anti-s, anti-D (IAT) 1. Cassettes BioVue® Anti-Human Globulin anti-IgG
2. Reagents
ORTHO™ Sera Anti-Fya (Anti-FY1) ORTHO™ Sera Anti-Fyb (Anti-FY2) ORTHO™ Sera Anti-S (Anti-MNS3) ORTHO™ Sera Anti-s (Anti-MNS4) ORTHO™ Sera Anti-D (IAT) (Anti-RH1)
3. Sample 4% patient red blood cells suspensions in saline (A6) or 0.8% patient red blood cells suspensions in saline (A7)
4. 4% Procedure + + + + + +
Identify the cassette Add 40 µl ORTHO Sera – P1 Add 10 µl 4% patient red blood cells suspensions – P2 15 minutes incubation 5 minutes centrifugation Reading (both sides)
5. 0.8% Procedure + + + + + +
Identify the cassette Add 40 µl ORTHO Sera – P1 Add 50 µl 0.8% patient red blood cells suspensions – P3 15 minutes incubation 5 minutes centrifugation Reading (both sides)
SpeCific Procedures - TYPING/SCREENING/CROSS-MATCH
B Page 25
B20 Extended Phenotyping – Anti-Jka, anti-Jkb, Anti-K, anti-D (DVI), anti-P1, anti- Lea, anti-Leb 1. Cassettes BioVue® Reverse Diluent (6 x Reverse)
2. Reagents
ORTHO™ Sera Anti-Jka (Anti-JK1) ORTHO™ Sera Anti-Jkb (Anti-JK2) ORTHO™ Sera Anti-K (Anti-KEL1) ORTHO™ Sera Anti-D (DVI) (Anti-RH1) ORTHO™ Sera Anti-P1 (Anti-P1PK1) ORTHO™ Sera Anti-Lea (Anti-LE1) ORTHO™ Sera Anti-Leb (Anti-LE2)
3. Sample 4% patient red blood cells suspensions in saline (A6) or 0.8% patient red blood cells suspensions in saline (A7)
4. 4% Procedure + + + + +
Identify the cassette Add 40 µl ORTHO Sera – P1 Add 10 µl 4% patient red blood cells suspensions – P2 5 minutes centrifugation Reading (both sides)
5. 0.8% Procedure + + + + +
B
Identify the cassette Add 40 µl ORTHO Sera – P1 Add 50 µl 0.8% patient red blood cells suspensions – P3 5 minutes centrifugation Reading (both sides)
SpeCific Procedures - TYPING/SCREENING/CROSS-MATCH
Page 26
B21 Extended Phenotyping – Anti-M, anti-N 1. Cassettes BioVue® Neutral
2. Reagents ORTHO™ Sera Anti-M (Anti-MNS1) ORTHO™ Sera Anti-N (Anti-MNS2)
3 Sample 4% patient red blood cells suspensions in saline (A6) or 0.8% patient red blood cells suspensions in saline (A7)
4 4% Procedure + + + + +
Identify the cassette Add 40 µl ORTHO Sera – P1 Add 10 µl 4% patient red blood cells suspensions – P2 5 minutes centrifugation Reading (both sides)
5 0.8% Procedure + + + + +
Identify the cassette Add 40 µl ORTHO Sera – P1 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 27
C1 Agglutination Grading Chart 4+
3+
Agglutinated cells form a band at the top of the bead column.
Most agglutinated cells remain in the upper half of the bead column.
2+ Agglutinated cells observed throughout the length of the bead column. A small button of cells may also be visible at the bottom of the bead column.
1+ Most agglutinated cells remain in the lower half of the bead column. A button of cells will also be visible at the bottom of the bead column.
Neg All cells pass through the beads and form a button at the bottom of the bead column.
C Page 28
ORTHO BIOVUE® RESULTS INTERPRETATION
C2 Difficulties Solving/Troubleshooting Problem
Bubble in the glass beads column
Before testing, liquid level above glass beads is low (