bioFILM PATM ANTIMICROBIAL SUSCEPTIBILITY
SYSTEM PROCEDURAL MANUAL
Version 15.0
bioFILM PATM ANTIMICROBIAL SUSCEPTIBILITY SYSTEM PROCEDURAL MANUAL, Version 15.0
Health Canada Medical Device Licence Number 78222
Table of Contents
INTENDED USE ...................................................................................................................................................... 3
SUMMARY AND PRINCIPLES .................................................................................................................................. 3
PRECAUTIONS ....................................................................................................................................................... 3
STORAGE ............................................................................................................................................................... 3
PRODUCT DETERIORATION ................................................................................................................................... 3
SPECIMEN COLLECTION, HANDLING, AND PREPARATION ..................................................................................... 3
MATERIALS PROVIDED AS KIT COMPONENTS: ...................................................................................................... 4
MATERIALS AND EQUIPMENT REQUIRED BUT NOT PROVIDED:............................................................................ 4
CATION ADJUSTED MUELLER-HINTON BROTH (CAMHB) PERPARATION ............................................................... 4
PROCEDURE OUTLINE ........................................................................................................................................... 4
LIMITATIONS ......................................................................................................................................................... 7
QUALITY CONTROL ................................................................................................................................................ 7
REFERENCES ........................................................................................................................................................ 10
©Innovotech, Inc., 2010
The Innovotech logo is a registered trademark of Innovotech Inc.
Page 2 of 10
Date of Issue: 9 March 2011
bioFILM PATM ANTIMICROBIAL SUSCEPTIBILITY SYSTEM PROCEDURAL MANUAL, Version 15.0
Health Canada Medical Device Licence Number 78222
INTENDED USE
bioFILM PA™ is a proprietary qualitative, in-vitro diagnostic test designed for use in determining antimicrobial
susceptibility of planktonic and biofilm Pseudomonas aeruginosa.(1)
SUMMARY AND PRINCIPLES
This broth dilution antimicrobial susceptibility test has various antimicrobial agents alone and in combination
which are diluted in Cation Adjusted Mueller-Hinton Broth (CAMHB). These antimicrobial agents are at categorical
breakpoint concentrations defined by the Clinical and Laboratory Standards Institute (CLSI) (2)(3). A standardized
suspension of P. aeruginosa from a clinical isolate is allowed to form biofilms on a 95-peg lid under incubation for
4-6 hours. Planktonic susceptibility and resistance are determined by measuring inhibition and growth in the
presence of antimicrobial agents after 16-24 hours of incubation. The biofilms are then placed in CAMHB, and
biofilm susceptibility and resistance are determined by measuring inhibition and growth in the absence of
antibiotics after an additional 16-24 hours of incubation.
PRECAUTIONS
 Single use only.
 Results should always be interpreted in conjunction with a patient’s medical history, clinical presentation, and
other findings.
 Laboratory technicians must be appropriately trained according to the bioFILM PA™ Antimicrobial Susceptibility
System Procedural Manual.
 Aseptic techniques and good laboratory practice should be observed throughout all procedures, with special
awareness that the plates and inoculation lid contain potentially pathogenic organisms.
 All waste including expired or unused product should be decontaminated by autoclave, incineration, or chemical
means prior to disposal in compliance with facility/institutional guidelines.
STORAGE
 Store Plate A (IPG1) and the rinse plate at ambient temperature.
 Store Plate B (IPA1) at -60oC or below
 Store Plate C (ICMAS1) at -20oC or below.
PRODUCT DETERIORATION
Prolonged exposure to storage conditions other than those recommended may result in loss of potency of the
antimicrobial agents contained in Plate B. Do not use the kits beyond their expiration date. Once thawing occurs,
plate B should be used within the same day or discarded. Plate C should be used within 24 hours or discarded.
SPECIMEN COLLECTION, HANDLING, AND PREPARATION
A clinical isolate of P. aeruginosa is required. Appropriate specimens should be collected, handled, transported,
and placed on primary isolation media according to procedures recommended in the Manual of Clinical
Microbiology (4).
If you are dealing with sputum samples, perform the following steps in Laminar air Flow Hood:
Using a sterile disposable loop, streak out the sputum sample on Pseudomonas Isolation HVEG agar.

Reclose the sputum container tightly and keep it at 4±1°C till bacterial growth is confirmed.

Incubate the agar plate in 35±1°C incubator for 16-24 hours. Some clinical strains might take longer

time to grow on agar plate (48 hrs).
Additionally, inoculate 150 mL of sterile Trypticase Soy broth directly from the sputum sample using a

sterile disposable loop and incubate in 35±1°C incubator for 16-24 hours. The overnight culture will be
used for subculturing on agar plates later on as an alternative step.
Discard the sputum samples appropriately.

Using the API 20 NE kit from Biomerieux is an optional recommended step for sputum or any other clinical
samples to confirm the identification of the bacterial isolates in your sample.
©Innovotech, Inc., 2010
The Innovotech logo is a registered trademark of Innovotech Inc.
Page 3 of 10
Date of Issue: 9 March 2011
bioFILM PATM ANTIMICROBIAL SUSCEPTIBILITY SYSTEM PROCEDURAL MANUAL, Version 15.0
Health Canada Medical Device Licence Number 78222
MATERIALS PROVIDED AS KIT COMPONENTS:
 Proprietary 95-peg inoculation lid with 96-well microtiter plate (IPG1), marked as Plate ‘A’
 Proprietary Antimicrobial challenge plate (IPA1), marked as Plate ‘B’
 Recovery plate (ICMAS1), marked as Plate ‘C’ (alternatively, plate of sterile Cation Adjusted Mueller Hinton
Broth can be prepared from powder according to the manufactures directions or purchased premade in
bulk from a commercial supplier)
 96-well microtiter rinse plate
MATERIALS AND EQUIPMENT REQUIRED BUT NOT PROVIDED:
 Quality Control organism Pseudomonas aeruginosa ATCC 27853
 Platform shaker (Set at 110 revolutions per minute(rpm))
 General laboratory equipment
 Single channel micropipette (approximately 40 µL and 300 µL required)
 Multichannel micropipette (150 µL and 200 µL required, 12 channels recommended)
 Sterile pipettes (3 mL and 19.7 mL required)
 Sterile glass test tube
 Sterile reservoir
 0.5 McFarland Barium Sulfate Turbidity Standard
 Sterile distilled deionised water
 Tryptic Soy Broth (TSB)
 Blood Agar (for primary and secondary subcultures)
 Pseudomonas aeruginosa isolation agar (Pseudomonas Isolation HVEG)
 API 20 NE kit (optional)
 Cation Adjusted Mueller-Hinton Broth (CAMHB)
 96 Well Nunc microtiter plate (e.g. clear flat-bottom Nunc MicroWell 96-well Microplates from Thermo
Scientific Catalogue number 167008)
 Non-selective media (eg. Tryptic Soy Agar ) for inoculum and purity checks
 Materials to perform an inoculum check using an appropriate procedure
 For mucoid strains: 0.1% Tween® 80 or Triton® X-100
 35±1°C incubator, non-CO2
 Spectrophotometer
 Materials that might be used in the serial dilution for inoculum check (e.g. 96 wells plate)
 Reporting forms
CATION ADJUSTED MUELLER-HINTON BROTH (CAMHB) PERPARATION
Follow the manufacturer’s procedures to prepare the broth from powder.
PROCEDURE OUTLINE
Turnaround time requires 24 hours for Minimum Inhibitory Concentration (MIC) results, and 48 hours for
Minimum Biofilm Eradication Concentration (MBEC) results.
A: INOCULUM PREPARATION: The turbidity standard technique(5) is recommended for direct inoculation of P.
aeruginosa.
A1. For Non-Mucoid Strains
a) Choose approximately 4-5 large or 5-10 small, well isolated P. aeruginosa colonies from an 18-24 hour
culture on blood agar.
b) Emulsify in 3 mL of sterile distilled deionised water in a sterile glass test tube and mix well. If required,
adjust to achieve a turbidity equivalent to a 0.5 McFarland standardized suspension. Alternatively,
measure the absorbance in a spectrophotometer at a wavelength of 625 nm, with water as a blank.
©Innovotech, Inc., 2010
The Innovotech logo is a registered trademark of Innovotech Inc.
Page 4 of 10
Date of Issue: 9 March 2011
bioFILM PATM ANTIMICROBIAL SUSCEPTIBILITY SYSTEM PROCEDURAL MANUAL, Version 15.0
Health Canada Medical Device Licence Number 78222
Adjust to achieve the expected absorbance between 0.08-0.13(6).
Other nephelometers could be used to check optical density. Some examples of are listed below.
McFarland Standard No.
1.0% Barium chloride (ml)
1.0% Sulfuric acid (ml)
Approx. cell density (1 X 108 CFU/mL)
% Transmittance at wavelength of 600 nm
Absorbance at wavelength of 600 nm
c)
0.5
0.05
9.95
1.5
74.3
0.132
1
0.1
9.9
3.0
55.6
0.257
2
0.2
9.8
6.0
35.6
0.451
3
0.3
9.7
9.0
26.4
0.582
4
0.4
9.6
12.0
21.5
0.669
Pipette 300 µL of the standardized suspension into 19.7 mL of TSB and mix well.
NOTE: Steps A1.b to B.a should be completed within 30 minutes.
A2. For Mucoid Strains
a) Choose approximately 3-5 well-isolated P. aeruginosa colonies from an 18-24 hour culture on blood agar.
b) Emulsify in 4-5 mL of TSB in a sterile tube, cap the tube and mix well. The solution may contain 0.1%
Tween® 80 or Triton® X-100 to assist in dispersion of microorganisms.
c) Loosen the cap and incubate overnight (in a non-CO2 incubator) at 35±1°C until the culture achieves or
exceeds the turbidity of a 0.5 McFarland standard.
d) Following overnight incubation: Mix well and adjust the turbidity with TSB in a sterile glass test tube to
achieve a turbidity equivalent to a 0.5 McFarland standardized suspension. Alternatively, measure the
absorbance in a spectrophotometer at a wavelength of 625 nm, with water as a blank. Adjust to achieve
the expected absorbance between 0.08-0.13(6).
Other nephelometers could be used to check optical density. Some examples of are listed below.
McFarland Standard No.
1.0% Barium chloride (ml)
1.0% Sulfuric acid (ml)
Approx. cell density (1 X 108 CFU/mL)
% Transmittance at wavelength of 600 nm
Absorbance at wavelength of 600 nm
0.5
0.05
9.95
1.5
74.3
0.132
1
0.1
9.9
3.0
55.6
0.257
2
0.2
9.8
6.0
35.6
0.451
3
0.3
9.7
9.0
26.4
0.582
4
0.4
9.6
12.0
21.5
0.669
e) Pipette 300 µL of the standardized suspension into 19.7 mL of TSB and mix well.
NOTE: Steps A2.d to B.a should be completed within 30 minutes.
B: INOCULATION OF PLATE A AND BIOFILM FORMATION
Always ensure that a lid and base are aligned correctly. The lid will have a tiny pin located in one of the
corners. A base will have diagonal cuts on two corners. A base will only align with a lid if the tiny pin is
aligned with one of the diagonal corner cuts of a base.
a)
Lift the peg lid from Plate A and inoculate with 150 µL of the inoculum (from step A1.c and/or A2.e) into
each well. This step should be completed within 30 minutes after preparing the inoculum from step A1.b
and/or A2.d.
b) Return the lid and check that it is aligned properly and fits securely.
c)
Place the inoculated Plate A onto a platform shaker set at 110 rpm. Incubate at 35±1°C in a non-CO2
incubator for 4-6 hours.
NOTE: This is sufficient for most isolates to generate a planktonic inoculum density that is approximately
5 x 105 CFU/mL, as recommended by the CLSI(5). However, some P. aeruginosa clinical isolates
may require extended or overnight incubation at this step for biofilm formation to meet CLSI
inoculum density recommendations.
OPTIONAL STEP: To confirm the growth rate of the Pseudomonas isolates, prepare a diluted
bacterial suspension by diluting 0.5 mL of the 0.5 McFarland standardized bacterial suspension
©Innovotech, Inc., 2010
The Innovotech logo is a registered trademark of Innovotech Inc.
Page 5 of 10
Date of Issue: 9 March 2011
bioFILM PATM ANTIMICROBIAL SUSCEPTIBILITY SYSTEM PROCEDURAL MANUAL, Version 15.0
Health Canada Medical Device Licence Number 78222
from step A.1.b or A.2.d, and place it in a sterile glass tube containing 2.0 mL of sterile TSB. This
suspension has OD625 of 0.05. Incubate this bacterial suspension in parallel with inoculated plate
A for 4-6 hours. Check the OD625 of the bacterial suspension in the glass tube. If OD625 falls
between 0.15-0.4, then start step C.a, otherwise leave plate A in the incubator for 18-20 hours.
d) Optionally, take 100 µL of the original bacterial suspension prepared in step A1.c or A2.e to perform an
inoculum check.
e) Remove Plate B and Plate C (if provided) from the freezer and allow equilibration to room temperature
for approximately 120 minutes. It is acceptable to store plate C (if provided) at room temperature until
used in step C.g. Alternatively prepare a fresh plate of Cation Adjusted Mueller Hinton Broth by adding
200 µL of prepared stock broth solution to each well of a 96 well microtiter plate (Nunc)
C: ANTIMICROBIAL SUSCEPTIBILITY TESTING
a) After incubation on the platform shaker, remove the lid from Plate A and transfer onto Plate B. Dispose
the base of Plate A.
b) Perform an inoculum check immediately. Use the inoculum fluid from the growth control well (G12) of
Plate B by using an appropriate procedure to ascertain whether the inoculum meets CLSI (5) requirements.
Remove up to 80 µL to allow for a minimum of 100 µL to remain in the well.
c) Prepare a purity plate by streaking a loopful of inoculum fluid from the growth control well (G12) onto
non-selective medium, incubate (in a non-CO2 incubator) for 16-20 hours. If a mixed culture is present on
the purity plate, re-isolation of test colonies and retesting is warranted.
d) Incubate the plate from step C.a) at 35±1°C in a non-CO2 incubator for 16-24 hours. Do not place the
plates on a platform shaker or stack them more than 4 high.
e) Remove Plate C from the freezer (if provided) and allow equilibration to room temperature for
approximately 120 minutes. Alternatively use the prepared plate of Cation Adjusted Mueller Hinton
Broth.
f) After incubation (step C.d.), immediately transfer the 95-peg lid onto a 96-well microtiter rinse plate
containing 200 µL of sterile distilled deionised water in each well. Leave for 30 seconds.
g) Transfer the 95-peg lid from the rinse plate onto Plate C. Discard the rinse plate.
h) Incubate this plate at 35±1°C in a non-CO2 incubator for 16-24 hours. Do not place the plates on a
platform shaker or stack them more than 4 high.
i) Use the base of Plate B (containing antibiotics) to read the Planktonic results within 30 minutes after
removal from the incubator (refer to Section D). Discard the base after reading.
j) After incubation (Step C.g), read the Biofilm results from Plate C within 30 minutes after removal from
the incubator (refer to Section D).
k) Decontaminate waste by autoclave, incineration, or chemical means prior to disposal in compliance with
facility/institutional guidelines.
D: READING THE PLATES
Plates can be read manually using a microtiter reading mirror. To receive Recording forms, contact your
Innovotech Inc. representative, or email Innovotech Inc. at biofilmpa@innovotech.ca.
Wipe the bottom of the plate with lint-free tissue to remove any condensation or debris that may be present.
 Growth in antimicrobial wells appears as turbidity, which may take the form of a white haze throughout the
well, and/or fine granular growth throughout the well.
 No growth is defined as a clear well or slight whiteness in the well or broth.
 Read the plates only if the Growth Control well (G12) is turbid and there is no growth in the Sterility Control
(H12) well.
 Do Not Read the plates if there is no growth in the Growth Control well (G12), or, if there is growth in the
Sterility Control well (H12); re-testing is warranted.
©Innovotech, Inc., 2010
The Innovotech logo is a registered trademark of Innovotech Inc.
Page 6 of 10
Date of Issue: 9 March 2011
bioFILM PATM ANTIMICROBIAL SUSCEPTIBILITY SYSTEM PROCEDURAL MANUAL, Version 15.0
Health Canada Medical Device Licence Number 78222
E: INTERPRETATION OF PLANKTONIC AND BIOFILM SUSCEPTIBILITIES
Susceptibility is determined by comparing the breakpoint susceptibility of an organism with either the
attainable blood or urine level of the antimicrobial agent. Table 1 lists the interpretive breakpoint criteria for
antibiotics used in the bioFILM PA™ kit.
 Growth in neither concentration of the antimicrobial should be recorded as susceptible (S).
 Growth in the lower concentration of the antimicrobial, but not in the higher concentration, should be
recorded as intermediate (I). Refer to the notes below:
 NOTE: There are no interpretive criteria for Intermediate values for the single antibiotics
Piperacillin/tazobactam (P/T) and Trimethoprim/sulfamethoxazole (T/S); therefore, growth in the lower
concentration of the antimicrobial, but not in the higher concentration, should be recorded as susceptible
(S). This also applies when these antibiotics are together in combination (T/S/P/T).
 NOTE: For combination antibiotics involving P/T and T/S, interpretive criteria of ‘Intermediate’ is not
applicable (unevaluable) when there is growth in the lower concentration of the antimicrobial, but not in
the higher concentration.
 Growth in both concentrations of the antimicrobial should be recorded as resistant (R).
 If there is growth in the higher concentration of the antimicrobial but not in the lower concentration, this is
considered a skipped well (5) and should be recorded as resistant (R).
LIMITATIONS
 Some slow growing P. aeruginosa strains may require extended or overnight incubation (at step B.c) for biofilm
formation to meet CLSI(5) inoculum density recommendations.
 Biofilm (MBEC) susceptibility results will be attained if the organism is able to form a biofilm.
 P. aeruginosa may develop resistance during prolonged therapy with all antibiotics. Therefore, isolates that
are initially susceptible may become resistant within three to four days after initiation of therapy(2). Testing
of repeat isolates may be warranted.
QUALITY CONTROL
The acceptability of the antimicrobial agents in the bioFILM PA™ Susceptibility Kit should be checked in
accordance with CLSI(2)(3) guidance by testing with Pseudomonas aeruginosa ATCC 27853(2)(3), refer to Table 2.
Reproducibility information and Quality Control procedures can be made available upon request from Innovotech
Inc.
©Innovotech, Inc., 2010
The Innovotech logo is a registered trademark of Innovotech Inc.
Page 7 of 10
Date of Issue: 9 March 2011
bioFILM PATM ANTIMICROBIAL SUSCEPTIBILITY SYSTEM PROCEDURAL MANUAL, Version 15.0
Health Canada Medical Device Licence Number 78222
Table 1: Interpretive Breakpoint Criteria – Unless otherwise noted, based on the MIC Interpretive Standards as
indicated in table 2B-1in CLSI Document M100-S20(1)(2). (Units: µg/mL)
Antimicrobial Agent
Susceptible Intermediate
Resistant
Amikacin
≤ 16
32
≥ 64
Aztreonam
≤8
16
≥ 32
Cefepime
≤8
16
≥ 32
Ceftazidime
≤8
16
≥ 32
Chloramphenicol(3)
≤8
16
≥ 32
Ciprofloxacin
≤1
2
≥4
Colistin
≤2
4
≥8
Gentamicin
≤4
8
≥ 16
Meropenem
≤4
8
≥ 16
Piperacillin/tazobactam
≤ 64/4
-
≥ 128/4
Trimethoprim/sulfamethoxazole(3)
≤ 2/38
-
≥ 4/76
≤4
8
≥ 16
Amikacin/aztreonam
≤ 16/8
32/16
≥ 64/32
Amikacin/cefepime
≤ 16/8
32/16
≥ 64/32
Amikacin/ceftazidime
≤ 16/8
32/16
≥ 64/32
Amikacin/ciprofloxacin
≤ 16/1
32/2
≥ 64/4
Amikacin/colistin
≤ 16/2
32/4
≥ 64/8
Amikacin/meropenem
≤ 16/4
32/8
≥ 64/16
Amikacin/piperacillin/tazobactam
≤ 16/64/4
-
≥ 64/128/4
Amikacin/trimethoprim/sulfamethoxazole(3)
≤ 16/2/38
-
≥ 64/4/76
Chloramphenicol (3)/ceftazidime
≤ 8/8
16/16
≥ 32/32
Chloramphenicol (3)/meropenem
≤ 8/4
16/8
≥ 32/16
Ciprofloxacin/aztreonam
≤ 1/8
2/16
≥ 4/32
Ciprofloxacin/colistin
≤ 1/2
2/4
≥ 4/8
Ciprofloxacin/meropenem
≤ 1/4
2/8
≥ 4/16
Ciprofloxacin/piperacillin/tazobactam
≤ 1/64/4
-
≥ 4/128/4
Ciprofloxacin/trimethoprim/sulfamethoxazole(3)
≤ 1/2/38
-
≥ 4/4/76
Gentamicin/aztreonam
≤ 4/8
8/16
≥ 16/32
Gentamicin/cefepime
≤ 4/8
8/16
≥ 16/32
Gentamicin/ceftazidime
≤ 4/8
8/16
≥ 16/32
Gentamicin/ciprofloxacin
≤ 4/1
8/2
≥ 16/4
Gentamicin/colistin
≤ 4/2
8/4
≥ 16/8
Gentamicin/meropenem
≤ 4/4
8/8
≥ 16/16
Gentamicin/piperacillin/tazobactam
≤ 4/64/4
-
≥ 16/128/4
Gentamicin/trimethoprim/sulfamethoxazole(3)
≤ 4/2/38
-
≥ 16/4/76
Tobramycin/aztreonam
≤ 4/8
8/16
≥ 16/32
Tobramycin/cefepime
≤ 4/8
8/16
≥ 16/32
Tobramycin/ceftazidime
≤ 4/8
8/16
≥ 16/32
Tobramycin/ciprofloxacin
≤ 4/1
8/2
≥ 16/4
Tobramycin/colistin
≤ 4/2
8/4
≥ 16/8
Tobramycin/meropenem
≤ 4/4
8/8
≥ 16/16
Tobramycin/piperacillin/tazobactam
≤ 4/64/4
-
≥ 16/128/4
Tobramycin/trimethoprim/sulfamethoxazole(3)
≤ 4/2/38
-
≥ 16/4/76
Trimethoprim/sulfamethoxazole(3)/aztreonam
≤ 2/38/8
-
≥ 4/76/32
Trimethoprim/sulfamethoxazole(3)/ceftazidime
≤ 2/38/8
-
≥ 4/76/32
Trimethoprim/sulfamethoxazole(3)/meropenem
≤ 2/38/4
-
≥ 4/76/16
≤ 2/38/64/4
-
≥ 4/76/128/4
Tobramycin
Trimethoprim/sulfamethoxazole(3)/piperacillin/tazobactam
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The Innovotech logo is a registered trademark of Innovotech Inc.
Page 8 of 10
Date of Issue: 9 March 2011
bioFILM PATM ANTIMICROBIAL SUSCEPTIBILITY SYSTEM PROCEDURAL MANUAL, Version 15.0
Health Canada Medical Device Licence Number 78222
Table 2:
Acceptable limits for the Quality Control of P. aeruginosa ATCC 27853 and expected QC endpoints for
Planktonic susceptibility in the bioFILM PA™ Kit - Unless otherwise noted, based on the acceptable
limits for the Quality Control of P. aeruginosa ATCC 27853 indicated in table 4 of CLSI Document M100S20(2).
Antimicrobial Agent
Amikacin (AK)
Aztreonam (AZT)
Cefepime (CPE)
Ceftazidime (CAZ)
Chloramphenicol (7) (C)
Ciprofloxacin (CP)
Colistin (CT)
Gentamicin (GM)
Meropenem (MER)
Piperacillin/Tazobactam (P/T)
Tobramycin (TO)
Trimethoprim/Sulfamethoxazole (T/S)
MIC acceptable limits
(µg/mL)
1–4
2–8
1–8
1–4
>16
0.25 – 1
0.5 – 4
0.5 – 2
0.25 – 1
1/4 - 8/4
0.25 – 1
8/152 – 32/608
Expected QC
Result
< 16
≤8
≤8
<8
> 16
≤1
≤4
<4
<4
< 32:4
<4
>2:38
Dilution(s) on Panel
16,32
8,16
8,16
8,16
8,16
1,2
2,4
4,8
4,8
32:4,64:4
4,8
1:19, 2:38
Due to the nature of the test, MIC values are of planktonic cells in the presence of biofilms. Biofilms constantly
release cells; therefore, although planktonic cells are killed, a constant supply of cells can be gained by the media
from the biofilm. This can result in slight whiteness in the wells that must be taken into account during quality
control. Refer to Tables 3 and 4 for a list of antibiotics where prominent growth and slight whiteness may or may
not be observed during quality control.
Table 3: List of Antibiotics and concentrations where prominent growth should always appear during quality
control.
Antibiotic(s)
T/S
T/S
C
C
Concentration (µg/mL)
2:38
1:19
16
8
Table 4: List of Antibiotics and concentrations where slight whiteness may or may not be observed during quality
control.
Antibiotic(s)
CAZ
CPE
CPE
MER
MER
P/T
AZT
AZT
Concentration
(µg/mL)
8
16
8
8
4
32:4
16
8
Antibiotic(s)
CT
CT
C/MER
C/MER
C/CAZ
C/CAZ
T/S/P/T
T/S/MER
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Concentration
(µg/mL)
4
2
16/8
8/4
16/16
8/8
1:19/32:4
1:19/4
Antibiotic(s)
T/S/AZT
T/S/AZT
T/S/CAZ
T/S/CAZ
Page 9 of 10
Concentration
(µg/mL)
2:38/16
1:19/8
2:38/16
1:19/8
Date of Issue: 9 March 2011
bioFILM PATM ANTIMICROBIAL SUSCEPTIBILITY SYSTEM PROCEDURAL MANUAL, Version 15.0
Health Canada Medical Device Licence Number 78222
REFERENCES
1. The BioFilm PA™ diagnostic test kit, components, methods of use, and test panel layout, are
protected by Innovotech, Inc. under one or more patents and/or pending patent applications. All
rights reserved.
2. M100 - S20 Performace Standards for Antimicrobial Susceptibility Testing; Twentieth Informational
Supplement. Wayne : Clinical and laboratory Standards Institute, 2010.
3. M100 - S16 Performance Standards for Antimicrobial Susceptibility Testing; Sixteenth Informational
Supplement. Wayne : Clinical and Laboratory Standards Institute, 2006.
4. Murray, P. R., et al. Manual of Clinical Microbiology. 9th Edition. Washington : American Society of
Microbiology, 2007.
5. M07- A8 Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria That Grow Aerobically;
Approved Standard - Eighth Edition. Wayne : Clinical and Laboratory Standards Institute, 2009.
6. Andrews, J. M. BSAC standardized disc susceptibility testing method (version 4). Journal of
Antimircobial Chemotherapy, 2005, 56 : 60-76.
7. MacGowan, A. P. and Wise, R. Establishing MIC breakpoints and the interpretation of in vitro
susceptibility tests. Journal of Antimicrobial Chemotherapy, 2001, 48 Supplement 1: 17-28.
For questions, concerns, and technical support, please contact Innovotech Inc.:
Mail:
Innovotech Inc. Suite 101, 2011-94 Street, Edmonton, Alberta, T6N 1H1, Canada
Phone:
(780) 448-0585
Fax: (780) 424-0941
Email:
biofilmpa@innovotech.ca
bioFILM PA™ Test Panel Layout (All units are expressed in g/mL)
GM/AZT
8/16
GM/MER
8/8
AK/AZT
32/16
AK/MER
32/8
TO/AZT
8/16
TO/MER
8/8
T/S/P/T
2:38/64:4
CP/AZT
2/16
CP/P/T
2/64:4
P/T
64:4
TO
8
CAZ
16
GM/AZT
4/8
GM/MER
4/4
AK/AZT
16/8
AK/MER
16/4
TO/AZT
4/8
TO/MER
4/4
T/S/P/T
1:19/32:4
CP/AZT
1/8
CP/P/T
1/32:4
P/T
32:4
TO
4
CAZ
8
GM/CAZ
8/16
GM/CP
8/2
AK/CAZ
32/16
AK/CP
32/2
TO/CAZ
8/16
TO/CP
8/2
T/S/MER
2:38/8
CP/CT
2/4
C/MER
16/8
AZT
16
T/S
2:38
CPE
16
GM/CAZ
4/8
GM/CP
4/1
AK/CAZ
16/8
AK/CP
16/1
TO/CAZ
4/8
TO/CP
4/1
T/S/MER
1:19/4
CP/CT
1/2
C/MER
8/4
AZT
8
T/S
1:19
CPE
8
GM/P/T
8/64:4
GM/T/S
8/2:38
AK/P/T
32/64:4
AK/T/S
32/2:38
TO/P/T
8/64:4
TO/T/S
8/2:38
T/S/AZT
2:38/16
CP/T/S
2/2:38
C/CAZ
16/16
CT
4
C
16
MER
8
GM/P/T
4/32:4
GM/T/S
4/1:19
AK/P/T
16/32:4
AK/T/S
16/1:19
TO/P/T
4/32:4
TO/T/S
4/1:19
T/S/AZT
1:19/8
CP/T/S
1/1:19
C/CAZ
8/8
CT
2
C
8
MER
4
GM/CPE
8/16
GM/CT
8/4
AK/CPE
32/16
AK/CT
32/4
TO/CPE
8/16
TO/CT
8/4
T/S/CAZ
2:38/16
CP/MER
2/8
AK
32
GM
8
CP
2
GC
GM/CPE
4/8
GM/CT
4/2
AK/CPE
16/8
AK/CT
16/2
TO/CPE
4/8
TO/CT
4/2
T/S/CAZ
1:19/8
CP/MER
1/4
AK
16
GM
4
CP
1
SC
Unless otherwise stated, breakpoint values for antibiotics adopted from MIC Interpretive Standards as indicated
in table 2B-1in CLSI Document M100-S20(2).
Antibiotics:
GM =
AK =
CAZ =
CPE =
T/S =
P/T =
gentamicin
amikacin
ceftazidime
cefepime
trimethoprim/sulfamethoxazole (3)
piperacillin/tazobactam
©Innovotech, Inc., 2010
The Innovotech logo is a registered trademark of Innovotech Inc.
AZT =
MER =
TO =
CP =
CT =
C=
aztreonam
meropenem
tobramycin
ciprofloxacin
colistin
chloramphenicol (3)
Page 10 of 10
GC = growth control
SC = sterility control
Date of Issue: 9 March 2011