Haemonetics eBDSTM

The total solution for detection of bacteria in all platelets and red cells

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Haemonetics eBDSTM: Operationally Proven to Make Bacteria Detection Simple and Effective

Features and Benefits

� Advanced technology designed to support blood bank cGMP. Easily integrates into routine practice.

� Unsurpassed sensitivity and specificity.

� Single use, closed system processing minimises false positives.

� Single point detection provides test results as early as 24 hours post collection.

System Components:

eBDS Sample Set

Sterile sample set, designed for easy sample transfer and oxygen sampling.

eBDS Oxygen Analyser Easy to use with cGMP functions. Modular to allow easy scalability. Small footprint.

Incubation and Agitation Platelet agitation accelerates bacteria growth and oxygen equilibration between the head space air and fluid.

Haemonetics Data Automatically and securely captures process data. Monitors and manages the entire eBDS process. Sends data to host IT by FTP.

The Haemonetics eBDS uses oxygen concentration as a marker for bacterial growth. It is intended for quality control and release testing of apheresis and whole blood-derived platelet products in plasma or platelet additive solutions (PAS) and for red cells in additive.

� Minimal component loss (4 mL).

� Integrated end to end IT system to document, manage and prompt entire sampling, detection and data output process.

� Now validated for red cells and non-leucocyte reduced platelets.

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1 3 4

2 min

A sample pouch is sterile connectedminimising the risk of contaminationand simplifying staff requirement.

Test eBDS pouches at any time from 24 hours after donation to out date.

Data is automatically captured, securely saved to database and may be further sent to the Central IT system by automated transfer.

Sterile connect the eBDS Sample Set to platelet product.

Incubate at 35 °C for 18-48 hours.

Read oxygen value using theeBDS Oxygen Analyser.2

Sample at any time from t=0 to out date (optimal sensitivity from 24 hours onward).

Fill sample pouch with 3 mL (approx) of platelet product.

eBDSTM Process Flow

4 simple steps to safe platelets. 4 minutes handling per platelet test.

� Provides the air for oxygen measurements in the sample pouch.

� Can be used for platelet count samples or stripped back to reduce component loss (approximately 4 mL).

� Prevents potential back flow of sample pouch contents to protect the platelet product.

� Slows flow into sample pouch to ease filling.

� Fill lines provide users with a visual stop for sample collection.

� Labelling tab provides an ISBT 128 bar code with unique lot numbers, for ultimate traceability, and an area to place unit identification on sample pouch, if required.

Integral Solution Option: Due to the dry nature of the eBDS pouch it can easily integrate into closed systems such as apheresis platelet kits or the Haemonetics AutoStop BC High Efficiency In-Process Filter System for Leucocyte Removal from Pooled Buffy Coat Platelets. This confers 3 additional benefits:

� Minimises handling.

� Reduces cost associated with sterile connection.

� Minimises the potential for sample contamination. Protected Sampling Port: � Allows access to the headspace air within the sample pouch.

� Contains a hydrophobic membrane that repels fluid away from the sampling site.

� Pre-slit injection site for easier insertion of the blunt probe of the eBDS Oxygen Analyser. Bacteria Growth Enhancing Tablets Contain: � Trypticase Soy Broth (TSB), a nutrient that enhances bacterial growth.

� Sodium Polyanethol Sulphonate (SPS), inhibits natural bactericidal agents in plasma and causes platelets to aggregate, reducing their oxygen consumption.

eBDS Sample Set

Sterile Dockable Tubing:

Check Valve:

Sample Pouch:

1 min1 min

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Incubator

� 2 sizes available to accommodate all levels of production. � Advanced electrical control systems assure accurate and consistent incubation temperature regardless of the atmospheric conditions (up to 40 °C).

� Integrated chart recorder is included in all models to provide continuous monitoring of incubation temperature for operational assurance.

� Rapid throughput - approximately 30 seconds to obtain reading - up to 40,000 tests per year.

� Automatic data entry - generates data string for electronic capture. � Simple Pass or Fail displays - no interpretation necessary. � Appropriate security levels - passcode entry required to

access set-up menus. � Requires no preventive maintenance - instrument is calibrated annually.

� Automatic hourly baseline confirmation - provides quality assurance.

� Four barcode input options - User ID, Lot Number, Product Code, Donation ID; any or all of the fields may be selected for use.

� Barcode symbology recognition - prevents incorrect barcode entry (ISBT 128, Codabar, USS Codabar).

� PC handshake feature verifies that a PC with data collection software remains connected to the analyser. An alarm will display if disconnected, ensuring data cannot be lost.

� Includes Oxygen Sampling Stand, a device that holds the sample pouch upright and secure during oxygen measurements.

� Two RS232 ports - for connection to a barcode scanner and computer.

eBDSTM Oxygen Analyser:

Components and Features:

The Testing Principle Behind the Haemonetics eBDS

Percent oxygen in air, the practical approach

The Haemonetics eBDS novel approach to detection measures

the oxygen content of air within the sample pouch as a marker for

bacteria growth. The eBDS Oxygen Analyser is used to measure

the percent oxygen in the sample pouch head space. If bacteria

are present in the platelet sample collected, oxygen is consumed in

the sample pouch head space through the metabolic activity

and proliferation of the bacteria.

Haemonetics Data offers a seamless, end to end IT solution for the management of the entire eBDS process from log in of the platelet donation ID and time through to automated data export to your host IT system. Capture, Workflow, Query, Interface, Watcher and Sign modules allow for secure data capture and ensure cGMP compliance. All modules are configurable to support 21 CFR part 11 if desired.

� Data is captured into a secure, industry standard Microsoft® Sequel ServerTM data base ensuring data integrity and facilitating real time secure access by multiple users

� “Haemonetics Data Workflow” module allows management, tracking and audit of the platelet unit hold at room temperature, sampling and 35 °C incubation including instant views of results as they are generated

� The “Query” module allows intuitive searches of any element of the database to view donation centric information, or search by sample ID, date/time, analyser serial number etc.

� Interface supports the management of data to the host IT, viewing records sent and resending data as required.

� Interface gives the laboratory manager the opportunity to hold back records from export to the host IT until they have been authorised. Audit functionality allows a view of who did what at every stage of the eBDS process. Product Related Threshold Haemonetics Data automatically captures and saves the component type: platelets in plasma, platelets in PAS (Platelet Additive Solution) or red cells and applies the correct oxygen Pass/Fail threshold for the Oxygen Analyser.

Haemonetics Data

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Table 2. Bacteria levels in platelet products at the time of inoculation and after 24-hour storage at which time samples were taken into theeBDS Sample Set (Sample Time = 24 hours), and the resulting detection frequency. Inoculation Inoculation Detection with bacterial level bacteria level Sampling at 24 hours

Median (range) Median (range) <5 <5 6 - 15 16 - 50 16 - 50 16 - 50 >51 >51 Cases detected Cases detected CFU/mL CFU/mL CFU/mL CFU/mL CFU/mL CFU/mL CFU/mL CFU/mL CFU/mL CFU/mL of cases of cases Plasma PAS Plasma PAS Plasma PAS Plasma PAS Plasma PAS sampled Plasma sampled PAS

S. epidermidis (ATCC 49134) 7 (2 - 52) 4 (1 - 10) 1 15 15 7 8 2 3 2 27 of 27 25 of 25

S. agalactiae (ATCC 12927) 5 (2 - 20) 10 (1 - 17) 3 6 7 2 9 8 9 10 28 of 28 26 of 26

S. aureus (ATCC 27217) 8 (2 - 51) 8 (3 - 25) - - - 2 5 - 24 24 29 of 29 26 of 26

P. aeruginosa (ATCC 27853) 9 (1 - 15) 8 (3 - 17) - 1 1 1 4 - 19 24 24 of 24 26 of 26

S. choleraesuis (ATCC 8326) 8 (1 - 55) 10 (2 - 34) 6 - 0 3 2 7 16 9 24 of 24 22 of 22

E. coli (ATCC 25922) 6 (2 - 15) 6 (1 - 20) - - - - - - 27 20 27 of 27 20 of 20

E. cloacae (ATCC 29005) 8 (2 - 13) 13 (5 - 32) 4 - 4 1 4 - 16 19 28 of 28 20 of 20

B. cereus (ATCC 7064) 13 (3 - 27) 3 (1 - 7) - - - - 2 - 31 20 33 of 33 20 of 20

K. pneumoniae (ATCC 8045) 5 (1 - 17) 5 (1 - 14) 12 - 9 1 3 2 9 17 33 of 33 20 of 20

S. marcescens (ATCC 43862) 9 (1 - 16) 9 (1 - 18) 2 - - - - - 25 20 27 of 27 20 of 20

Total 28 24 36 17 37 19 179 165 280 of 280 225 of 225 (100%) (100%)

Bacteria level immediately after inoculation and sampling Detection with (Sample Time = 0 hours) Sample Time = 0 hours

<5 <5 6 - 15 6 - 15 16 - 50 16 - 50 Cases detected Cases detected CFU/mL CFU/mL CFU/mL CFU/mL CFU/mL CFU/mL of cases of cases Plasma PAS Plasma PAS Plasma PAS sampled Plasma sampled PAS

S. epidermidis (ATCC 49134) 4 17 1 9 - - 5 of 5 25 of 26

S. agalactiae (ATCC 12927) 5 7 4 18 2 1 11 of 11 25 of 26

S. aureus (ATCC 27217) - 6 5 17 4 3 9 of 9 26 of 26

P. aeruginosa (ATCC 27853) - 5 11 17 - 4 8 of 11 12 of 26

S. choleraesuis (ATCC 8326) 4 2 2 18 5 6 11 of 11 26 of 26

E. coli (ATCC 25922) 1 8 6 14 - 2 7 of 7 19 of 20

E. cloacae (ATCC 29005) - 1 6 14 - 5 6 of 6 20 of 20

B. cereus (ATCC 7064) 2 15 6 2 4 - 12 of 12 16 of 17

K. pneumoniae (ATCC 8045) 3 12 7 8 1 - 11 of 11 17 of 20

S. marcescens (ATCC 43862) - 9 5 10 - 1 5 of 5 20 of 20

Total 19 82 53 123 16 22 85 of 88 206 of 227 (96.6%) (90.7%)

Performance Summary for the Haemonetics eBDSTM

The ten organisms that were selected for determining the performance characteristics of the Haemonetics eBDS were the same genus and species as those responsible for over 98% of all fatalities associated with the transfusion of bacteria contaminated platelets1. The exact slow growing organisms isolated from platelet components were used rather than similar fast growing isolates ensuring real world reliability. The organisms were tested in platelet products prepared from whole blood-derived platelet rich plasma, buffy coat and apheresis. A similar performance study was conducted to validate the testing of red cells.

Data is shown and available from validation studies in both plasma and platelet additive solution (PAS)2-12. Haemonetics eBDS is approved for all routinely used PAS and plasma platelet components and leucocyte reduced red cells stored in SAG-M13–14.

Samples were collected using the eBDS Sample Set immediately after inoculation (Table 1), and after a 24 hour incubation following inoculation (Table 2). Platelet products were inoculated prior to using the eBDS Sample Set. Sample collection into the pouch was followed by 24 hour incubation at 35 °C and thereafter tested.

Sensitive*

� 96.6% (plasma) and 90.7% (PAS) of all samples tested positive when sampling occurred immediately following inoculation.

Table 2 illustrates that:

� 100% of all samples tested positive when sampling occurred after a 24 hour incubation at room temperature.

An independent study7 has shown eBDS to be of equivalentsensitivity to an established culture based method in a side-by-sidecontrolled study.

Specific

No false positives were detected in the control samples. Routine usemonitoring of the 118,067 Haemonetics eBDS test resulted in only 76 (1/1554 or 0.06%) false positive tests. This confirms the high specificity in a routine environment2.

Bacteria level at sampling time after 24 hours storage (Sample Time = 24 hours)

Table 1. Bacteria levels in the platelet products and detection with sampling from the platelet products performed immediately after inoculation (Sample Time = 0 hours).

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Bacteria level in whole blood-derived leuco-reduced red cell components at sampling time after 24 hours storage (Sample Time = 24 hours)

Table 3. Bacteria levels in the leuco-reduced red cell components and detection after 24 hour storage at which time samples were takeninto the eBDSTM Sample Set (Sample Time = 24 hrs), and the resulting detection frequency Detection with Sample Time = 24 hours

<5 6 - 15 16 - 50 >51 Cases detected CFU/mL CFU/mL CFU/mL CFU/mL of cases Plasma Plasma Plasma Plasma sampled Plasma

K. pneumoniae (ATCC 8045) 2 4 9 3 18 of 18

S. liquefaciens (ATCC 35551) 9 5 1 - 15 of 15

P. aeruginosa (ATCC 27853) 1 9 6 2 18 of 18

P. putida (ATCC 492819128) 1 2 - 3 6 of 6

P. fluorescens (ATCC 17569) 2 7 2 3 14 of 14

E. amnigenes (ATCC 33731) 6 1 2 - 9 of 9

E. coli (ATCC 25922) - 8 7 - 15 of 15

Y. enterocolitica (ATCC 27729) 9 1 2 5 17 of 17

B. cereus (ATCC 7064) - 4 3 5 12 of 12

L. monocytogenes (ATCC 19115) 3 1 6 - 10 of 10

S. aureus (ATCC 27217) - 9 1 - 10 of 10

S. epidermidis (ATCC 49134) 4 5 1 3 13 of 13

Total 37 56 40 24 157 of 157

(100%)

Footnote: Further data on testing of red cells at additional time points (0, 7, 21, 35-42 days) are available on request.

Red Cell Component Data

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The high sensitivity of eBDS allows for flexible sampling schedules to fit with different blood centre operational logistics:

� Sampling of the platelet unit after 24 hours hold followed by incubation of the eBDS pouch at 35 °C for 18-30 hours.

� Sampling of the platelet unit immediately after collection/preparation followed by incubation of the eBDS pouch at 35 °C for 24-30 hours.

� Red cells may be sampled as soon as 24 hours post donation. The eBDS pouch should be incubated for 48-72 hours.

eBDSTM Sampling Schedule Chart for Platelets

eBDS Sampling Schedule Chart for Leucocyte Reduced Red Cells in Additive Solution

eBDS 100% Sensitivity at 1-15 CFU/mL in plasma

eBDS 100% Sensitivity at 1-15 CFU/mL in red cells

Hold - Room Temp 35 °C Incubation

35 °C Incubation

T = 0 hr T = 24 hr T = 42 hr T = 54 hr Up to day 5/7

Up to day 5/7T = 0 hr T = 24 hr T = 30 hr

Blood Collection

Sample Detect 18 hr

Detect 30 hr

Detect 24 hr

Detect 30 hrSample

eBDS 100% Sensitivity at 1-15 CFU/mL in red cells

Hold - Room Temp 35 °C Incubation

T = 0 hr T = 24 hr T = 72 hr T = 96 hr Up to day 42

Blood Collection

Sample Detect 48 hr

Detect 72 hr

Ordering Information

† Includes 4 probes and filters. The information in this document should not be used as a substitute for the instructions for use of the eBDS Oxygen Analyser and the eBDS Sample Set.

© 2007, 2013 Haemonetics Corporation. Haemonetics, Haemonetics The Blood Management Company, and eBDS are trademarks or registered trademarks of Haemonetics Corporation in the USA, other countries, or both. All rights reserved. Microsoft is a registered trademark of Microsoft Corporation. Sequel Server is a trademark of Microsoft Corporation. 06.2013 USA. COL-COPY-000624-IE(AA)

Haemonetics S.A. Signy Centre Rue des Fléchères P.O. Box 262 1274 Signy Centre Switzerland www.haemonetics.com

Information contained herein for distribution outside of the USA only.

References 1. Lee JH. In: Bacterial Contamination of Platelets Workshop, Bethesda, September 24, 1999. (Cited May 3, 2000). Available from http://www.fda.gov/cber/minutes/bact092499.pdf. 2. Holme, S. Bunch, Selman B. (2005). Bacterial contamination in stored platelets: Performance of the Pall eBDS System under routine use conditions: Vox Sanguinis, 89 (suppl. 1), P-194. 3. Pearce S, Colley L, Taylor J, Rowe G, Wilkins K. (2004). Evaluation of Pall Bacteria Detection System (Pall eBDS). Vox Sanguinis, 87 (suppl. 3), A5.7. 4. Holme S, McAlister M, Ortolano G, Cortus MA, Jacobs M, Yomtovian R, Freundlich L. (2004). New system enhancements of a bacteria detection system (eBDS) based on oxygen consumption. Vox Sanguinis, 87 (suppl. 3), Tu03.04. 5. McDonald C, Colvin J, Wilkins K, Macdonald R, Fernandez M, Robbins S, Barbara J. (2004). Evaluation of the Pall enhanced Bacterial Detection System (eBDS) for testing platelet concentrates. Vox Sanguinis, 87 (suppl. 3), P5.7. 6. Holme S, McAlister M, Ortolano G, Chong C, Cortus M, Jacobs M, Yomtovian R, Freundlich L, Wenz B. (2005). Enhancement of a culture-based bacterial detection system (eBDS) for platelet products based on measurement of oxygen consumption. Transfusion, 45(6), 984-93. 7. McDonald C, Pearce S, Wilkins K, Colvin J, Robbins S, Colley L, Taylor J, Barbara JBJ. (2005). Pall eBDS: an enhanced bacterial detection system for screening platelet concentrates. Transfusion Medicine, 15(4), 259-68. 8. Fournier-Wirth C, Deschaseaux M, Defer C, Godreuil S, Carriere C, Bertrand X, Tunez V, Schneider T, Coste J, Morel P. (2006). Evaluation of the enhanced bacterial detection system for screening of contaminated platelets. Transfusion, 46(2), 220-4.

Reorder Code Description Pkg

400-03E eBDSTM Sample Set 40/per case†

9. D’Antonio D, Accorsi P, Balbinot A, Lacone A. (2004). Validation of Pall eBDS with platelets stored in additive solution. Transfusion Medicine and Hemotherapy, 31 (suppl. 3), PO1603. 10. Toscano L, Hansen A, Nielsen M, Arpi M, Grunnet N. (2004). Evaluation of an enhanced bacterial detection system in leucocyte depleted platelets store in platelet additive solution. Transfusion, 44 (suppl.), SP43. 11. McDonald C, Sapsford J, Colvin J, Biggs C, Goodhand K. (2006). Are the BACT/ALERT and Pall enhanced Bacterial Detection Systems suitable for screening platelet additive solution (PAS) platelet concentrates? Vox Sanguinis, 91 (suppl. 3), P-149. 12. Schmidt M, Karakassopoulos A, Burkhart J, Deitenbeck R, Asmus J, Müller TH, Weinauer F, Seifried E and Walther-Wenke G. (2007). Comparison of three bacterial detection methods under routine conditions. Vox Sanguinis, 92, 15-21. 13. McDonald C, Goodhand K, Wilkins K, Colvin J. (2006). Detection of bacterial contamination in red cell units using the Pall enhanced Bacterial Detection System. Vox Sanguinis, 91, (suppl. 3), P-148. 14. Chen CL, Yu J-C, Holme S, Jacobs MR, Yomtovian R and MacDonald C. (2008). Detection of bacteria of stored RBC products using a culture based bacterial detection system (eBDS). Transfusion, 48(8), 1550-1557.