Automate the

®
T-SPOT .TB test
Agenda
Automating the T-SPOT.TB process

T-Cell Select™ reagent kit

Validated magnetic cell separation systems

Other automation solutions to complete the T-SPOT.TB test process

Why you should automate the T-SPOT.TB test

 Automation for every step

Complete solutions for full 2 day workflow available

Liquid handling steps of the plate development process can be automated

Each result should be individually validated before reporting
 T-Cell Select reagent kit
CE marked reagent kit to automate the isolation of mononuclear immune
cells from peripheral blood samples
Complete kit of reagents for use in cell-mediated immune assays:

Kit components:

Reagent Quantity
Buffer Concentrate 1 x 50 mL
Antibody 1 3 x 2 mL
Antibody 2 3 x 2 mL
Bead Reagent 1 x 10 mL
• All required reagents are included in the kit
• Each kit supports up to 144* tests
o Up to 6 T-SPOT.TB test kits

*Actual number of tests will vary based on laboratory techniques and instrument solution
The T-Cell Select reagent kit replaces density
gradient centrifugation

• Reagents for density gradient centrifugation are no longer required:

⚬ T-Cell Xtend® reagent
⚬ Centrifuge tubes
⚬ Density gradient centrifugation materials:
⎻ Leucosep™ tubes
⎻ Centrifuge tubes + Ficoll-Paque® media
⎻ Cell Preparation Tubes (CPT ®)

• Media used for cell washing:

⎻ RPMI 1640 and AIM-V ® are still required but in lower volumes
Manual method comparison to the automated cell
separation workflow
Leucosep tube manual cell separation

The T-Cell Select reagent kit cell separation

Add Buffer, add Antibody 1 Place sample plate on
Incubate Add Bead Reagent cell separation system
+ Antibody 2
How does the T-Cell Select reagent kit work?

Magnetic cell separation of mononuclear immune cells

• Magnetic beads and antibodies target surface markers of specific cell types
• These specific cell types are isolated directly from the peripheral blood
using a magnet
• Positive selection is used to ensure high recovery and purity

Over 500,000 T-SPOT.TB test results have been reported using

magnetic bead based cell isolation at Oxford Diagnostic Laboratories®
The positive selection process using an
in-tube magnet
2. Blood incubation with
buffer, antibodies and beads
4. Magnetic retention
of bead-bound cells
from blood sample
5. Selected cells
washed multiple
3. Magnetic collection times in RPMI.
of bead-bound cells
from blood sample
1. Whole blood
sample transfer to
tubes / deep well plate

6. Elution of bead selected

cells in AIM-V
Enhanced sample requirements

Lower blood volumes required but performance maintained

• Magnetic cell separation is more efficient than Blood volume required

density gradient centrifugation Immunocompetent adults 3.5 mL
• Comparable cell yield despite lower blood Immunosuppressed adults 2 x 3.5 mL
requirement Children >2 years 3.5 mL
• Standard heparin or citrate tubes are used Children <2 years 2.0 mL
• No special blood collection tubes required

3.5 mL 2 mL

Different separation systems may require different blood volumes - volume must be validated by the laboratory
 Strong concordance between methods

95 % confidence
Agreement excluding interval
borderline results:
Lower Upper
Overall agreement 97.3 % 95.5 % 98.2 %

• Clinical studies compared the T-SPOT.TB test results with

cells isolated using: 95 % confidence
Agreement including interval
⚬ T-Cell Select reagent kit (0-54 hour sample storage) borderline results:
Lower Upper
⚬ Density gradient centrifugation and T-Cell Xtend reagent
Overall agreement 94.2 % 92.3 % 95.8 %
(0-32 hour sample storage)

Oxford Immunotec package insert: PI-TS-IVD-UK-V5 10

Improved sample stability, improved logistics

• Blood samples processed up to 54 hours post-venepuncture

⚬ Maximum time to start processing using T-Cell Xtend reagent is 32 hours

• Greater lab flexibility

⚬ Batch testing possible & late arriving samples can be stored overnight for next day processing

• Room temperature sample storage

⚬ No need for refrigerated sample transport, specialized tubes or complex sample manipulation

• Expand the reach of your laboratory

⚬ Samples can be shipped from further away or different courier routes can be explored
 Magnetic bead-based cell separation systems
Validated cell separation systems validated for use with the T-Cell Select
reagent kit
• The following Allsheng and Thermo Scientific™ systems have been fully validated, and are
supported, by Oxford Immunotec:

Allsheng systems
• Auto-Pure10B Thermo Scientific systems
• Auto-Pure20B
• KingFisher™ Flex
• Auto-Pure24

If other systems than those named above are used, these must be validated in your own lab and our customer support team will
not be able to support implementation and may not be able to trouble shoot
 Allsheng systems
The Allsheng family of magnetic cell separation systems

Auto-Pure20B Auto-Pure10B Auto-Pure24

~45-minute
processing
time

• For 1-20 samples • For 1-10 samples • Up to 24 samples per run

• Cartridge type • Cartridge type • Deep Well Plate (DWP)
 Display screen:

Auto-Pure20B (AP20B)
Features
• Capable of processing 1 to 20 samples per run
• Consumables: specifically designed tip combs
and 7-tube strips
• UV sterilisation – to avoid cross contamination

Inside the AP20B:

Benefits
✓ Simple to use and maintain with an easy-to-
use touch screen
✓ Flexible batch run
✓ Small footprint Tube strips:
• 30 kgs: 400 mm x 520 mm x 450 mm
(W×D×H)

* Oxford Immunotec has validated the Auto-Pure20B & Auto-

Watch the AP20B in action Pure24 with an elution volume of 550μL. Depending on cell count
methods and counters used later in the T-SPOT process, an
alternative elution volume in the range 550μL – 1000μL can be
validated if required.
 Display screen
Auto-Pure10B (AP10B)
Features
• Capable of processing 1 to 10 samples per run
• Consumables: specifically designed tip comb and
7-tube strips
• UV sterilisation – to avoid cross contamination

Benefits Magnetic rod tips

✓ Simple to use and maintain with an easy-to-use touch
screen
✓ Flexible batch run Tube strips:
✓ Small footprint Blood
+
Master Mix RPMI *
• 18 kgs: 340 mm x 350 mm x 410 mm (W×D×H) (700 μL)
(3.5 mL) (580 µL) (1 mL each)

* Oxford Immunotec has validated the Auto-Pure10B with an elution volume of

700μL. Depending on cell count methods and counters used later in the
T-SPOT process, an alternative elution volume in the range 550μL – 1000μL
can be validated if required.
 Display screen
Auto-Pure24 (AP24)
Features
• Capable of processing up to 24 samples per run
• Consumables: unique 24 deep well plate (DWP) for samples
and reagents and magnetic rod tip cover
• Footprint
• 54 kgs: 50 mm x 620 mm x 483 mm (W x D x H) Magnetic rod tips:

Benefits
✓ Can be integrated into a fully automated workflow with liquid
handler
✓ Good collection of bead-bound sample due to the effectiveness DWPs:
of the magnet
✓ Low maintenance
Inside the AP24:

4 DWPs:
1. Elution (AIM-V)
2. Wash (RPMI)
Watch the AP24 in action 3. Blood sample
4. Tip comb (for magnetic rod tips)
The KingFisher Flex (KFF)
(Thermo Fisher Scientific) Display screen

Features
• Up to 24 samples per run
• Consumables: 24 deep well plate (DWP) for samples and reagents
and magnetic rod tip
• Footprint:
• 28 kgs, 680 mm x 600 mm x 380 mm (W x D x H)
Magnetic rod tips:
DWP:
Benefits
✓ Can be integrated into a fully automated workflow with liquid
handler
✓ Easy to use software
✓Reputable brand – good global service coverage (outsourced)

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 Cell separation systems: feature comparison
Instrument Maximum Plastic Compatible Waste Ready to Sample System
samples ware with liquid (with part use with LIS throughput Cost
per run handling? run)
Auto-Pure10B 10 Cartridge Not currently + Not currently + +
Auto-Pure20B 20 Cartridge Not currently + Not currently ++ +
Auto-Pure24 24 DWP ✓ ++ Not currently +++ ++
KingFisher Flex 24 DWP ✓ ++ ✓ +++ +++

Instrument dimensions: AP10B AP20B AP24 KFF

 Auto-Pure10B, 20B, 24 and KingFisher Flex
example workflow

• Auto-Pure10B,20B, 24 & KingFisher Flex automate the cell isolation step of the T-SPOT.TB test process on Day 1
• Automated options are available for other steps of the T-SPOT.TB process (examples above)
• Instrument run time is approximately 45 minutes

Day 1 & 2 of the T-SPOT.TB process (using above workflow)*

* Based on 24 sample batch size. Additional gains with smaller 10 or 20 sample run

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Cell separation systems:
customer feedback
✓ No technical issues
✓ Extremely easy to use
✓ Excellent support and training by OI
• Key benefits:
✓ More walk-away time
✓ Simple to use
✓ Process standardization for
accreditation

Satisfied customers

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 Automation for every step

Other automation solutions to complete the T-SPOT.TB test process

Liquid handling steps of the plate development process can be automated

Each result should be individually validated before reporting
Cellaca™ MX
(Nexcelom)
Features
• High throughput automated cell counter
• Plate-based for up to 24 samples
Counting plates (2x12)
• Small footprint: Laptop included

• 19kgs, 330 mm x 330 mm x 410 mm

Benefits
✓ High speed sample processing – can count 24 samples in
less than 3 minutes
✓ Easy to use, low cost and minimal consumable MATRIX software
requirements
✓ Minimal maintenance, OI installable and part of the
PerkinElmer company
Classification: RUO, users must validate the
performance of the system according to local
Click here to see unboxing and getting started with requirements

Cellaca MX Image above based on 3x8 plate orientation

Guava® easyCyte™ 5HT & Guava® Muse®
(Luminex)
Guava easyCyte 5HT
Features
• Automated cell counting
• Guava easyCyte 5HT: High-throughput, up to 96 samples with ~25-
minute run time for 24 samples
• Guava Muse: low throughput, single loader, <2 minutes per sample

Guava Muse
Benefits
✓ Counting with industry-leading precision
✓ Guava easyCyte 5HT: can be integrated into a fully automated
workflow with liquid handler
✓ Guava Muse: easy to use, compact and portable analyser

Watch the Guava Muse in action

 Janus G3
(PerkinElmer)
Features
• Liquid handler for the T-SPOT.TB test Display screen
(In development)
• Automates liquid handling steps for
Day 1 and 2
• Up to 48 samples per run
Benefits
✓ Enables hands-off preparation/processing of
samples for T-SPOT procedures
✓ Reduces human-error
✓ Better sample reproducibility

For more information on how a liquid handler could

optimize your T-SPOT.TB workflow, please contact one
of our automation specialists 24
Microwell Imager
(EUROIMMUN)
Features
• Automated spot visualization aid
• Plug and play system design
• Small footprint benchtop device:
• 13.6 kgs, 190 mm x 310 mm x 400 mm
Display screen

Benefits
✓ Fast run time of <2 mins
✓ Easy to use and low cost
✓ Minimal maintenance, OI installable and part of the
PerkinElmer company
BIOREADER® 7000-E
(BIO-SYS)
Features
• Automated plate reader with simple functionality
• High throughput capability and flexible, can do 1 sample
or 24
• Has software to support technologist in interpretation of
results Display screen

Benefits
✓Customer specific export and report templates
✓ > 300 Bioreader® units sold worldwide
✓ LIMS/LIS import and export capabilities
CTL S6 TATC
(CTL ImmunoSpot®)
Features
• Automated plate reader with simple functionality
• High throughput capability and flexible, can do 1 sample or 24
• Has software to support technologist in interpretation of results

Benefits
✓Good user interface and easy to use
✓LIMS/LIS export capabilities
✓Globally recognised with good service cover in different regions
 Choose your perfect level of automation

Modular systems to precisely fit your laboratory needs

Liquid handling steps of the plate development process can be automated

Each result should be individually validated before reporting
Time comparison for 2 day process

Reduce hands-on time by up to 60%

• Validated automation solutions are

available for the 2 day work flow
• Hands-on time reduced
significantly
• The automated 2 day T-SPOT.TB
workflow requires less hands-on
time than a typical automated
ELISA protocol

Standard ELISA protocol - Dynex DS2® run with 22 samples. Automated T-SPOT.TB process run with 24 samples
Hands-on time includes any period interacting with the process plus any walk away time less than 15 minutes
Timings can vary depending on equipment, throughput and operator speed
Automate your T-SPOT.TB test today
The most sensitive and specific test for TB infection is now automated 1,2

The T-SPOT.TB test, can now:

✓ Improve your workflow efficiency

✓ Reduce your hands-on time
✓ Extend your sample stability by up to 54 hours

…with modular automation solutions to fit your needs.

We can arrange for an automation specialist to visit your

lab to discuss your requirements and to work out the
best solutions for you and your team.
 Thanks for your
attention
CPT is a trademark of Becton Dickinson
FICOLL and FICOLL-PAQUE are registered trademarks of GE Companies
Dynex DS2 is a registered trademark of Dynex Technologies
Leukosep is a trademark of Greiner Bio-One international
Any Questions? Thermo Scientific and KingFisher are trademarks of Thermo Scientific
Allsheng is a trademark of Hangzhou Allsheng Instruments CO., Ltd
AIM-V is a registered trademark of Life Technologies Corporation
Cellaca is a trademark of Nexcelom Bioscience LLC, a PerkinElmer company.
Guava and Muse are registered trademarks of Luminex Corporation
easyCyte is a trademark of Luminex Corporation
The Janus is a registered trademark of PerkinElmer Inc
BIOREADER is a registered trademark of Biosys GmbH
ImmunoSpot is a registered trademark of Cellular Technology Limited
References: T-Cell Select is a trademark of Oxford Immunotec
1 Lewinsohn DM, Leonard MK, LoBue PA, et al. Official American ThoracicSociety/Infectious T-SPOT, T-Cell Xtend, Oxford Diagnostic Laboratories and the Oxford Immunotec logo are
Diseases Society of America/Centers for Disease Control and Prevention Clinical Practice registered trademarks of Oxford Immunotec
Guidelines: Diagnosis of Tuberculosis in Adults and Children. Clin Infect Dis. Published online © 2023 Oxford Immunotec. All rights reserved.
December 8, 2016:ciw694. doi:10.1093/cid/ciw694
TB-UK-PRES-MPN670-0001 V2
2 T-SPOT.TB package insert - PI-TB-IVD-UK V4
Extra slides if required

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The T-Cell Select reagent kit - summary

Extended sample stability gives you: Automation gives you:

1. Flexibility to batch run samples 1. Reduced hands-on time

2. Simple, room temperature, sample transport 2. More flexible, streamlined workflow

3. Time to ship samples from further away 3. Scalability to fit changes in throughput
requirements
 Positive selection - overview

Add buffer to the Add Expose samples to Remaining sample

whole blood sample Add antibodies to a magnetic field: Cells are washed,
Superparamagnetic discarded while
to facilitate cell bind to the Bead-antibody the magnet is
beads: complexes targeted cells are
purification and mononuclear complexed cells removed and the
form with the held in place by the
reduce red blood immune cells migrate to the cells are re-
antibodies attached magnet - positive
cell contamination magnet suspended
to immune cells selection

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The T-Cell Select reagent kit

Allows your lab to:

AUTOMATE the T-SPOT.TB test workflow
• Improve workflow
efficiency
• Streamline lab process
• Simplify and extend
sample delivery

Process samples up to 54 hours

post-venepuncture

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