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PROCEDURE
STORAGE OF ISOLATES
AND BLOOD CULTURE
BROTH
SOP-Bacterio-042-02
Page 1/4
Procedure: Storage of Bacterial isolates and Blood Culture
Broth
Implementation
Date
Author Signature Date
Name: Birgit De Smet
February 2013
Function:
Approval Signature Date
Name: T. Syna
Function
Changes Version 01 New
Version 03 February 2013
Purpose
The purpose of this procedure is to describe how to store:
• Bacterial isolates from positive blood cultures
• Blood culture broth from positive blood cultures
This procedure is used for clinical significant
organisms only NOT for contaminating bacteria
(coagulase negative staphylococci and Bacillus sp. do
not need to be stored )

PROCEDURE
STORAGE OF ISOLATES
AND BLOOD CULTURE
BROTH
SOP-Bacterio-042-02
Page 2/4
Remark
1. Store isolates and broth on the day of final reporting
2. Isolates are stored from fresh (overnight) and
pure culture plates
Materials
1. TSA tube (2 ml) for storage of isolate at room
temperature
2. BHI tube with glycerol and pearls (2 ml) for
storage of isolate at -20°C or -70°C (depend on
availability in lab)
3. Sterile tube (2ml) for storage of positive culture
broth
4. Sterile loop (for transfer of isolate to TSA tube)
5. Sterile swab (for transfer of isolate to BHI tube)
6. Sterile syringe + needle (for transfer of culture
broth to sterile tube)
7. 4 blank stickers to write (for numbers –check
Inventory Excel file):
o box number
o isolate number
o place in the box
o Stickers are for the 3 different tubes (TSA, BHI and
Broth) and 1 for the worksheet
8. Transparent tape (to cover the stickers on the
tubes)

PROCEDURE
STORAGE OF ISOLATES
AND BLOOD CULTURE
BROTH
SOP-Bacterio-042-02
Page 3/4
Procedure
for storage
of bacterial
isolate
1. Aseptically transfer a loop full of colonies from a pure,
fresh culture to a TSA tube – stab vertically into the
agar – close the tube very well and store in box
2. Take a sterile swab en take growth from a pure, fresh
culture – mix well in BHI tube – close the tube very well
and store in box
3. For culture broth: take the bottle - disinfect the rubber
stopper with Betadine solution and leave to dry for
2 minute - with sterile syringe and needle, take 1,5 ml
out of the bottle - Transfer to the cryotube and close the
cryotube - Place tube in box at -20°C or 70°C
Procedure
for
registration
1. Open the excel file “Inventory isolates and culture
broth”
2. Fill in 1 new row for the TSA and BHI tube in the Excel
sheet “Inventory isolates TSA and BHI”: complete
all column: Identification, patient number, blood culture
register number, date of collection blood for culture.
The column “Note”: you can use for special remark
3. Fill in 1 new row for the storage of culture broth in the
Excel sheet “Inventory culture broth”: complete all
column.
4. Save the file