EXAMINATION OF BACTERIAL

CONTAMINATION IN BLOOD COMPONENTS

Rain Lim
Customer Support Team Lead
ASEAN Industrial Microbiology PIONEERING DIAGNOSTICS
rain.lim@biomerieux.com
 BACTERIAL CONTAMINATION -
CLINICAL RELEVANCE
Most bacterial species can readily proliferate in the
nutrient-rich blood product environment during storage.
• Estimated level of contamination at the time of collection
is relatively low
- 1–10 colony forming units/mL or

Bacteria can proliferate within hours to reach levels of

106/mL or greater
- Such quantities of bacteria infused with the
transfusion product over a short period of time can
cause bacteremia and progress to sepsis and death.

Outcome of contaminated transfusion is highly dependent

on
• Amount of bacteria transfused
• Type of bacteria
• Pathogenicity of bacteria
• Rate of transfusion
• Clinical status of the recipient

Even healthy individuals can have a rapidly fatal outcome

when transfused with a large load of endotoxin-producing
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Gram-negative bacteria
 BLOOD BANK PRODUCTS –
PROCESSING

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 BACTERIAL CONTAMINATION -
SOURCES

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 MEASURES TO REDUCE BACTERIAL
CONTAMINATION

Donor questionnaire

Standardized skin disinfection (2004)

Diversion pouches that discard the 1st 30ml of

blood (2006)

Component testing is done in closed systems

(2006)

Bacterial contamination screening (2008)

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 Measures to reduce contamination-
Process Workflow
Discard
Donor Blood NO NO
Testing 1 Cleared? Stored
Screening Donation Sample

Day 0 YES

Testing 1 Processing Segment

/ Sampling Storage Testing 3
Blood group
Syphilis
Day 1
PRISM – HIV, HCV, HBV
NA Testing – HIV, HCV, HBV Testing 2

Testing 2 Cleared?
NO
Culture Test – BacT/Alert
YES
Cleared?
Testing 3

Gram Stain Reporting –

Culture Test – Bacteriology YES False
Positive

Dispatch
Sample /
Reporting
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 Measures to reduce contamination-
Donor Questionnaire

Pre-donation selection of donors

– Defer donors with dental treatment, skin infection,
diarrhea, colonoscopy and fever
– Travel to danger zone for the past few months
(e.g.Zika)

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 Measures to reduce contamination-
Skin disinfection

Donor skin disinfection

Iodine or Chlorohexidine with 70% spirit for at least
30 seconds

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 Strategies to reduce contamination-
Skin Disinfection

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 Measures to reduce contamination-
Blood Collection – Day 0
Removal of first aliquot of donor blood
– test tube or diversion pouch
– Blood group, Syphilis, PRISM, NAT, Archive

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 Measures to reduce contamination-
Collection/Storage Bags
Single Bag - Designed for the collection and storage of whole blood.

Double Bag - For the collection and storage of blood.

• Separates whole blood into two parts Eg red cells and plasma

Triple Bag - For the collection and storage of blood.

• Separates whole blood into different components.

Quadruple Bag - For the collection and storage of blood.

• Separate whole blood into different components. Eg red cells, buffy coat and
plasma

Apheresis – Automated collection of single donor Platelet and plasma

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 Measures to reduce contamination-
Collection/Storage Bags
Reducing packaging exposure
– 4 buffy coat + 1 plasma seed = 1 pooled platelet

Platelet
Use for
BTA 3D
Testing

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 MEASURES TO REDUCE CONTAMINATION-
BACTERIAL CONTAMINATION SCREENING

• Australia introduced bacterial contamination screening

(BCS) for platelet components in April 2008.

• 2010 to 2012, 1.1% of platelet donations tested initial

machine positive (IMP); since 2013 this rate has fallen to
0.6% through improved instrument management,
reducing false-positive IMPs but maintaining sensitivity
for cultures yielding bacterial growth.

• Incidence of reported transfusion-transmitted bacterial

infection (TTBI) has fallen since the introduction of BCS,
with a 4.2 fold lower rate from platelets and 2.8-fold
lower rate from red blood cells.
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 BacT/Alert Installed Base
(>1000 systems worldwide)

EUROPE

NORTH AMERICA

ASIA PACIFIC
Singapore
Australia
LATIN AMERICA Indonesia
Thailand

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 Bacterial testing of platelets

• Samples are pooled from 1 plasma + 4 pooled platelets (pool of

buffy coats with plasma seed, spin down)

• 5ml each into a set of aerobic and anaerobic bottle

• Surface sterilization (ethanol) are used prior to injection into BTA

bottles. (Ensure ethanol is vaporized from surface)

• Implicated product and sample segment are kept for further testing
(if required)

Pool by batch of 4 Inoculate into BTA bottles Segment for confirmatory test
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 Bacterial testing of platelets

BTA bottles in BTA 3D, 37oC for 5 days

If “+”

Perform culture and bacteria identification

- Implicated product
- Segment
- BTA bottles

Notification to be send and recall product

Discard

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 Measures to reduce contamination-
Post-transfusion Notification

Transfusion stopped immediately

Bacterial culture of patient

Broad spectrum antibiotic

Hae-movigilant report

Investigate for source of bacterial contamination

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 MICROBIOLOGICAL CONTROL MANAGEMENT
PHARMACEUTICAL INDUSTRY = BLOOD BANKS

ID & Susceptibility, Library

Final Product Monitoring

No Bacterial Contamination Sources

Donor Bacteremia Collection Processing

Blood pack fault monitoring
Beckman et al., Vox Sang (2004) 87, 272–279
“Severe transfusion-transmitted bacterial

Corrective Staphylococcus spp

Teeth Streptococcus viridans
Serratia liquefaciens
complications have been traced to leaks”
AABB Ass. Bull. #05-2, Jan 24, 2005
“It is important to identify the manner in which Yes Sampling
Action
Yersinia enterocolitica
Intestines Salmonella spp
Campylobacter spp
E. coli, Enterobacter spp.
Klebseilla, Serratia, etc.
Skin disinfection
• surface disinfection
bacteria entered the platelet unit (e.g. if the
source of contamination is related to a defect in
the transfusion set).”
TRANSFUSION, 2006-Vol. 46S, SP303
BUGS? Program
Needed? Osteomyelitis Staphylococcus
S. cholera suis
Several structures
• Sebaqeous glands, hair follicles
Scars
Eczema
Cameron et al.
“Serratia Liquefaciens Caused Hemagglutination
of Red Blood Cells”. “this bacterium has not been
Introduction of needle isolated since January 2006 after improved
• Small core punctured cleaning practices were implemented”
• runs with first 10 cc
Detection of bacterial contamination in
apheresis platelet products: American Red
Staphylococcus epidermidis
Staphylococcus aureus
Cross experience
Diptheroids spp Fang et al, TRANSFUSION 2005;45:1845-1852.

Yes Micrococcus spp

Pseudomonas spp
Bacillus cereus
Propionibacterium acnes
Flavobacterium spp
“Bacillus species was identified from one of the
confirmed cases. A pin hole, however, was found
in this particular PLT bag”. “it was suspected that
the contamination was introduced during storage”
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Process Monitoring
•In-Process
•Environment

Correct & Document

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 PLATELET CONCENTRATES
IDEAL SOLUTION?

Screen for Bacterial Contamination...

•Improve Safety of Platelet Supply
•Decrease Transfusion reactions, Mortality Rate
… and Increase Shelf-life
•Decrease Outdated stock
•Improve Supply (Weekends - Holidays)
•Bacterial Screening is proven to be Economically
very beneficial for the bloodbanker

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 BIOMÉRIEUX’S PORTFOLIO
TOTAL MICROBIOLOGICAL MANAGEMENT

• In-Process Control / Environmental Testing

– BacT/ALERT, air IDEAL, Count-Tact, manual sterility testing
media

• 100% Final Product Testing (Platelets, cord blood, stem cells,

bone/tissue) & In-Process Testing
– BacT/ALERT 3D

• ID for Root-Cause Analysis

• API Manual
• Vitek 2
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• Vitek MS
PIONEERING DIAGNOSTICS