WEEK 3: BACTERIOLOGY SECTION LEC 2: BLOOD CULTURE SYSTEM
BACT/ALERT 3D PROCEDURE
INTENDED USE:
 BACT/ALERT 3D
o Blood
o Sterile body fluids
PRINCIPLE
 BACT/ALERT Microbial Detection System utilize
a colorimetric sensor and reflected light to
monitor the presence and production of carbon
dioxide (CO2) that is dissolved in the culture
medium. If microorganisms are present in the
test sample, carbon dioxide is produced as the
organisms metabolize the substrates in the
culture medium. When growth of the
microorganisms produces CO2, the color of the
gas-permeable sensor installed in the bottom of
each culture bottle changes to yellow.
MATERIALS
 BACT/ALERT CULTURE BOTTLES
 BACT/ALERT 3D SYSTEM MACHINE
 
 
 
 
 
 
 
1. Remove the flip-off cap from the Bact/Alert vial
top and inspect the vials for cracks
contamination, excessive cloudiness and bulging
or indented stoppers. (do not use if any defect is
noted.)
2. Before inoculating, swab the septum with alcohol
(iodine is not recommended). Aseptically inject
or draw the required amount of blood directly.
3. Inoculated aerobic vials should be placed in the
Bact/Alert instrument as soon as possible for
incubation and monitoring. If placement of
theinoculated vial into the instrument has been
delayed and visible growth is apparent, it should
not be tested in the Bact/Alet instrument. It
should be subcultured, gram stained and treated
as presumptive positive.
4. Vial entered into the instrument will be
automatically tested every 10 minutes for the
duration of testing. Positive vials will be
determined by the Bact/Alert instrument and
identified as such. The sensor inside the bottle
will not appear visibly different in positive and
negative vials; The Bact/Alert however can
determine differencein fluorescene.
5. If at the end of the testing period, a negative vial
appears visibly positive (i.e., chocolatized blood,
bulging septum, lyzed and/or much darkened
blood) it should be subcultured, gram stained
and treated as presumptive positive.
6. Positive vials should be subcultured on BAP,
MacConkey, Mannitol salt and Chocolate agar.
INTERPRETATION
 Positive or negative culture bottles are
determined by decision making software
contained in Bact/Alert microbial detection
system. No action is required until the Bact/Alert
instrument signals culture bottles positive or
negative.