510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION

DECISION SUMMARY

A. 510(k) Number:

K163563

B. Purpose for Submission:

To obtain a substantial equivalence determination for Gentamicin for testing of gram

negative bacilli on the VITEK® 2 and VITEK® 2 Compact Antimicrobial Susceptibility Test
(AST) Systems

C. Measurand:

The VITEK® 2 AST-Gram Negative card contains the following concentrations of

Gentamicin: 4, 8, and 32 µg/mL (equivalent standard method concentration by efficacy in
µg/mL). The Gentamicin MIC reporting range for the card is ≤ 1 – ≥16 μg/mL.

D. Type of Test:

Automated quantitative or qualitative antimicrobial susceptibility test for Gentamicin

E. Applicant:

bioMérieux, Inc.

F. Proprietary and Established Names:

VITEK® 2 AST- GN Gentamicin (≤ 1 – ≥ 16 μg/mL)

G. Regulatory Information:

1. Regulation section:

21 CFR 866.1645: Fully Automated Short-Term Incubation Cycle Antimicrobial

Susceptibility System

2. Classification:

Class II

Page 1 of 12
 3. Product code(s):

LON – Fully automated short-term incubation cycle antimicrobial susceptibility system

LTW – Susceptibility Test Cards, Antimicrobial
LTT – Panels, Test, Susceptibility, Antimicrobial

4. Panel:

Microbiology (83)

H. Intended Use/Indications for Use:

1. Intended Use (s):

The VITEK® 2 Antimicrobial Susceptibility Test (AST) is intended to be used with the
VITEK® 2 Systems for the automated quantitative or qualitative susceptibility testing of
isolated colonies for the most clinically significant aerobic gram-negative bacilli,
Staphylococcus spp., Enterococcus spp., Streptococcus spp. and clinically significant
yeast.

2. Indications for Use:

VITEK® 2 AST-Gram Negative Gentamicin is designed for antimicrobial susceptibility

testing of Gram negative bacilli and is intended for use with the VITEK® 2 and
VITEK® 2 Compact Systems as a laboratory aid in the determination of in vitro
susceptibility to antimicrobial agents. VITEK® 2 AST-Gram Negative Gentamicin is a
quantitative test. Gentamicin has been shown to be active against most strains of the
microorganisms listed below, according to the FDA label for this antimicrobial.

Active in vitro and in clinical infections

Citrobacter species
Enterobacter species
Escherichia coli
Klebsiella species
Proteus species
Serratia species
Pseudomonas aeruginosa

The VITEK® 2 Antimicrobial Susceptibility Test (AST) is intended to be used with the
VITEK® 2 Systems for the automated quantitative or qualitative susceptibility testing of
isolated colonies for the most clinically significant aerobic gram-negative bacilli,
Staphylococcus spp., Enterococcus spp., Streptococcus spp. and clinically significant
yeast.

Page 2 of 12
 3. Special conditions for use statement(s):

For Prescription Use Only

Limitations:
“The ability of the VITEK 2 AST card to detect resistance with the following
combination(s) is unknown because resistant strains were not available at the time of
comparative testing:
Gentamicin: Proteus vulgaris”

“The ability of the VITEK 2 AST card to detect resistance with the following
combination(s) is unknown because an insufficient number of resistant strains were
available at the time of comparative testing:
Gentamicin: Citrobacter koseri, Enterobacter aerogenes, Enterobacter cloacae,
and Serratia marcescens”

4. Special instrument requirements:

VITEK 2 and VITEK 2 Compact Systems using VITEK 2 Systems 8.01 software

I. Device Description:

The VITEK 2 AST card is a miniaturized, abbreviated and automated version of the doubling
dilution technique for determining the minimum inhibitory concentration (MIC). Each
VITEK 2 AST card contains 64 wells. A control well(s) which contain only nutrient medium
is resident on all cards. The remaining wells contain premeasured portions of antimicrobials
combined with the nutrient media. The isolate to be tested is diluted to a standardized
concentration with 0.45% to 0.50% saline before being used to rehydrate the antimicrobial
medium within the card. The VITEK 2 System will automatically dilute the bacterial
suspension to prepare an inoculum for susceptibility cards. Then the VITEK 2 will fill, seal
and place the card into the incubator/reader. The VITEK 2 Compact has a manual filling,
sealing and loading operation. The VITEK 2 Systems monitor the growth of each well in the
card over a defined period of time (up to 24 hours for Streptococcus species). The analysis
program determines when a well demonstrates growth based on attenuation of light measured
by an optical scanner. This data is used to determine the minimum inhibitory concentration
or “MIC” values for the anti-microbial agent. At the completion of the incubation cycle, a
report is generated that contains the MIC value along with the interpretive category result for
each antimicrobial contained on the card.

VITEK 2 AST-GN Gentamicin has the following concentrations in the card: 4, 8, and 32
µg/mL (equivalent standard method concentration by efficacy in µg/mL). The MIC result
range for the VITEK 2 is ≤ 1 – ≥ 16µg/mL.

Page 3 of 12
J. Substantial Equivalence Information:

1. Predicate device name(s):

VITEK 2 AST-GN Doxycycline

2. Predicate 510(k) number(s):

K121546

3. Comparison with predicate:

Table 1: Comparison with Predicate Device

Similarities
Device: Predicate Device:
Item VITEK 2 AST-GN Gentamicin VITEK 2 AST-GN Doxycycline
(K163563) (K121546)
The VITEK 2 Antimicrobial
Susceptibility Test (AST) is intended to
be used with the VITEK 2 Systems for
the automated quantitative or qualitative
susceptibility testing of isolated
Intended Use Same
colonies for the most clinically
significant aerobic gram-negative
bacilli, Staphylococcus spp,,
Enterococcus spp., Streptococcus spp.
and clinically significant yeast.
Automated quantitative antimicrobial
susceptibility test for use with the
Test Method VITEK 2 and VITEK 2 Compact Same
Systems to determine the in vitro
susceptibility of Gram negative bacilli
Standardized saline suspension of test
Inoculum Same
organism
VITEK 2 Gram Negative Susceptibility
Test Card Same
Test Card
VITEK 2 and VITEK 2 Compact
Instrument Same
Systems

Page 4 of 12
 Differences
Device: Predicate Device:
Item VITEK 2 AST-GN Gentamicin VITEK 2 AST-GN Doxycycline
(K163563) (K121546)
Antimicrobial
Gentamicin Doxycycline
Agent
Antimicrobial
4, 8, 32 µg/mL 1, 4, 16 µg/mL
Concentrations
Reporting Range ≤ 1 - ≥ 16 μg/mL ≤ 0.5 - ≥ 16 µg/mL
Unique for gentamicin (Growth pattern Unique for doxycycline (Discriminate
Analysis Algorithm
analysis) analysis)

K. Standard/Guidance Documents Referenced (if applicable)

· FDA Class II Special Controls Guidance Document: Antimicrobial Susceptibility Test

(AST) Systems; Guidance for Industry and FDA (Issued August 28, 2009)

· CLSI M07-A9, “Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria
that Grow Aerobically; Approved Standard-Ninth Edition” Vol. 32 No. 2 (January 2012)

· CLSI M100-S24, “Performance Standards for Antimicrobial Susceptibility Testing”;

Twenty-fourth Informational Supplement, Vol. 33 No. 1 (January 2014)

L. Test Principle:

The VITEK 2 and VITEK 2 Compact Systems utilize automated growth-based detection
using attenuation of light measured by an optical scanner. The optics in the systems uses
visible light to directly measure organism growth within each of the 64 micro-wells.
Transmittance optics is based on an initial light reading of a well before significant growth
has begun. Every 15 minutes throughout the incubation cycle (defined period of time based
on the VITEK 2 card), light transmittance readings of each well measures organism growth
by the amount of light that is prevented from passing through the well. At the completion of
the incubation period, the MIC values and their associated interpretive category results for
each antimicrobial on the test card are displayed in an automatically generated report.

M. Performance Characteristics (if/when applicable):

1. Analytical performance:

a. Precision/Reproducibility:

A reproducibility study for the VITEK 2 AST-GN card with Gentamicin was
conducted at three clinical sites using ten isolates of gram-negative bacilli consistent
with the Intended Use. Testing was performed on three separate days and in triplicate
for a total of 270 data points. The isolates tested in the reproducibility study included
K. pneumoniae pneumoniae (five isolates), Citrobacter braakii (two isolates),
Citrobacter freundii (one isolate), and Pseudomonas aeruginosa (two isolates).

Page 5 of 12
 Inocula were prepared using both the auto-dilution and manual dilution methods for
testing in the VITEK 2 System. Inocula were prepared by the manual dilution method
only for use with the VITEK 2 Compact. The mode MIC value was determined and
the reproducibility was calculated based on MIC values that fell within +/- one
doubling dilution from the mode MIC value.

For VITEK 2 auto-dilution and manual dilution methods, overall reproducibility was
100% for best- and worst-case scenarios.

For the VITEK 2 Compact manual dilution method, overall reproducibility was 100%
for best- and worst-case scenarios.

The combined reproducibility results for all three sites were acceptable.

b. Linearity/assay reportable range:

Not applicable

c. Traceability, Stability, Expected values (controls, calibrators, or methods):

Inoculum Density Control

The DensiCHEK Plus was used to standardize the inoculum to a 0.5 McFarland
standard. The instrument was standardized daily with all results recorded at each site.
Calibration values were within the expected range.

Purity Check
A purity check of all organisms was performed on the dilution tube used to prepare
the VITEK 2 card inoculum. Only those cultures that were pure were evaluated in the
study.

Growth Failure Rate

There was one isolate that failed to grow in the VITEK 2 card in the clinical study.
Complete test results were available for 872 isolates in a total of 873 clinical isolates.
The growth failure rate was 0.1% and was acceptable.

All 75 challenge organisms grew in the VITEK 2 GN card with Gentamicin using
both the auto-dilution and manual dilution methods for the VITEK 2 and manual
inoculation for the VITEK 2 Compact System.

A total of 947 VITEK 2 AST results were available.

Quality Control (QC) Testing

The FDA/CLSI recommended QC organisms (E. coli ATCC 25922 and
Pseudomonas aeruginosa ATCC 27853) were tested using both the VITEK 2 card
and the reference method at each site. Both the automatic dilution and manual
dilution methods were used for the VITEK 2 and the manual dilution method was

Page 6 of 12
 used for the VITEK 2 Compact.

Both the auto-dilution and the manual dilution methods for VITEK 2 and the manual
dilution for VITEK 2 Compact were within the expected range >95% of the time.

Table 2: Quality Control Summary Results for VITEK 2 (Auto-Dilution and Manual Dilution
Methods) and VITEK 2 Compact (Manual Dilution Method)
Gentamicin VITEK 2 Auto- VITEK 2 VITEK 2 Compact
Dilution Manual Dilution Manual Dilution
Conc.
Organism Reference Test Reference Test Reference Test
(µg/mL)
≤0.0625
0.125
0.25
E. coli 0.5 214 105 105
ATCC 25922
(≤)1* 11 225 7 112 7 112
Expected Range
2
0.25 – 1 µg/mL
4
(VITEK 2:
≤1µg/mL) 8
16
32
≥64

≤0.0625
0.125
P. aeruginosa 0.25
ATCC 27853 0.5 35 17 17
Expected Range ≤1* 180 216 85 103 85 102
0.5 – 2 µg/mL 2 5 5 5 1
(VITEK 2: 4
≤1- 2µg/mL) 8 4 4 4
16
32
≥64
*The lowest dilution of the VITEK 2 Gentamicin MIC range is ≤1 µg/mL. Obtaining this value was considered an
indicator that the quality control test results were acceptable.

The Gentamicin expected ranges for E. coli ATCC 25922 and P. aeruginosa ATCC
27853 are 0.25 – 1 and 0.5 - 2µg/mL respectively. However, the VITEK 2 MIC
reporting range is ≤1 – ≥16 µg/mL (MIC results: ≤1, 2, 4, 8, ≥16 µg/mL). The VITEK
2 systems do not provide results lower than 1 µg/mL. Therefore, all results for E. coli
ATCC 25922 and P. aeruginosa ATCC 27853 were off scale.

A MIC value of ≤1 µg/mL indicated that the quality control test results were
acceptable.

bioMérieux included the following footnote to the QC table in the device labeling:

Page 7 of 12
 “The VITEK 2 Gram Negative Gentamicin does not include the full CLSI/FDA-
recommended dilution ranges for QC testing with E. coli ATCC 25922 and P.
aeruginosa ATCC 27853”.

d. Detection limit:

Not applicable

e. Analytical Specificity:

Not applicable

f. Assay cut-off:

Not applicable

2. Comparison studies:

a. Method comparison with predicate device:

Results obtained with the bioMérieux VITEK 2 AST-Gram Negative card with
Gentamicin were compared to results obtained with the CLSI broth microdilution
reference panel. The following concentrations of Gentamicin are contained in the
VITEK 2 AST-GN test card: 4, 8, and 32 µg/mL (equivalent standard method
concentration by efficacy in µg/mL) and the reporting range is ≤ 1 – ≥ 16 µg/mL (i.e.,
≤1, 2, 4, 8, and ≥16). The reference panel contained two-fold serial dilutions with a
range of ≤ 0.0625 to ≥128 µg/mL. The testing conditions for the reference method
consisted of the following:

· Medium – Mueller Hinton broth with appropriate dilutions of antimicrobial

solution added
· Inoculum – Direct colony suspension
· Incubation – 35°C ambient air incubator; 16-20 hours

All test inocula used for the evaluation of VITEK 2 AST-GN Gentamicin and the
reference method were standardized using the DensiCHEK Plus instrument. The
cards were inoculated with each test organism by auto-dilution for reading by the
VITEK 2 System and by manual dilution for reading on the VITEK 2 and VITEK 2
Compact Systems. Reference broth microdilution panels were inoculated in
adherence with CLSI document, M07-A9.

A total of 872 clinical isolates were evaluated at three sites with VITEK 2 AST –
Gram Negative cards inoculated by automatic dilution and interpreted using the
VITEK 2 instrument. The majority of isolates were fresh (760 isolates, 87.1%); 113
isolates (13%) were stock isolates.

Page 8 of 12
 A total of 75 challenge organisms (51 Enterobacteriaceae and 26 P. aeruginosa)
were evaluated at one site. The challenge set was tested with both the auto-dilution
and manual dilution options of the VITEK 2 System and with the manual dilution
method on the VITEK 2 Compact System. Overall performance of the challenge
organisms is shown in Table 2.

Table 2: Performance of Clinical and Challenge Isolates, VITEK 2 Auto-Dilution Method‡

Organism EA EA EA Eval EA Eval Eval CA CA
#R Min Maj Vmj
Group Total N % Total EA N EA % N %
Enterobacteriaceae ≤4 (Susceptible), 8 (Intermediate), ≥16 (Resistant)
Clinical 725 720 99.3 16 11 68.8 719 99.2 44 5 0 1
Challenge 51 51 100 4 4 100 50 98.0 10 1 0 0
Combined 776 771 99.4 20 15 75.0 769 99.1 54 6 0 1
Pseudomonas aeruginosa ≤4 (Susceptible), 8 (Intermediate), ≥16 (Resistant)
Clinical 147 146 99.3 46 45 97.8 141 95.9 12 6 0 0
Challenge 24 24 100 6 6 100 24 100 9 0 0 0
Combined 171 170 99.4 52 51 98.1 165 96.5 21 6 0 0
Enterobacteriaceae + Pseudomonas aeruginosa
Clinical 872 866 99.3 62 56 90.3 860 98.6 56 11 0 1
Challenge 75 75 100 10 10 100 74 98.7 19 1 0 0
Combined 947 941 99.4 72 66 91.7 934 98.6 75 12 0 1
‡
EA – Essential Agreement (+/- 1 doubling dilution) Min – Minor discrepancies
CA – Category Agreement Maj – Major discrepancies
EVAL – Evaluable isolates Vmj – Very major discrepancies
R – Resistant isolates
Essential Agreement (EA) occurs when there is agreement between the result of the reference method and that of VITEK 2
test card within plus or minus one serial two-fold dilution of the antibiotic. Evaluable results are those that are on scale for
both the VITEK 2 test card and the reference method. Category Agreement (CA) occurs when the interpretation of the
result of the reference method agrees exactly with the interpretation of the VITEK 2 test card.

The overall performance using the auto-dilution method of the VITEK 2 System
demonstrated an essential agreement of 99.4% and an overall category agreement of
98.6%. There was one very major (1.3%), 12 minor (1.3%) and no major
discrepancies. The Vmj was caused by K. pneumoniae.

The performance based on combined clinical and challenge data was acceptable.

There were no resistant Proteus vulgaris isolates tested in the studies. A limitation was
included in the package insert:
“The ability of the VITEK 2 AST card to detect resistance with the following
combination(s) is unknown because resistant strains were not available at the time of
comparative testing:
Gentamicin: Proteus vulgaris”

In addition, an insufficient number of resistant isolates were tested during the

comparative study for the following organisms: Citrobacter koseri (1), Enterobacter
aerogenes (1), Enterobacter cloacae (2), and Serratia marcescens (1). This was
addressed by adding the following limitation in the package insert:
“The ability of the VITEK 2 AST card to detect resistance with the following
combination(s) is unknown because an insufficient number of resistant strains were

Page 9 of 12
 available at the time of comparative testing:
Gentamicin: Citrobacter koseri, Enterobacter aerogenes, Enterobacter cloacae,
and Serratia marcescens”

Challenge Data – Auto and Manual Dilution:

The challenge set of 75 isolates (51 Enterobacteriaceae and 24 Pseudomonas
aeruginosa) was evaluated on the VITEK 2 System and the VITEK 2 Compact System
with the manual dilution method only.

Overall performance is shown in Table 3.

Table 3: Performance of Challenge Isolates, VITEK 2 and VITEK 2 Compact-Manual Dilution

Organism EA EA EA Eval EA Eval Eval CA CA
#R Min Maj Vmj
Group Total N % Total EA N EA % N %
VITEK 2 System
Auto-
75 75 100 10 10 100 74 98.7 19 1 0 0
dilution
Manual
75 75 100 10 10 100 74 98.7 19 1 0 0
dilution
VITEK 2 Compact
Manual
75 75 100 10 10 100 74 98.7 19 1 0 0
dilution

The performance of the challenge isolates using the VITEK 2 Gentamicin test on both
the VITEK 2 and VITEK 2 Compact Systems was acceptable.

MIC Trends:
The claimed Enterobacteriaceae organisms were also evaluated for trending. This
trending calculation takes into account MIC values that are determined to be ≤1 and
≥1 doubling dilutions compared to the reference method irrespective whether the
device MIC values are on-scale or not. The analysis showed trending was observed for
Proteus spp. and K. pneumoniae. The trending analysis is shown in Tables 4.1 and
4.2:

Table 4.1: Trending Analysis of Evaluable Clinical and Challenge

Results for Proteus spp.
Totala Difference in MIC as Compared to the CLSI Reference
Gentamicin (Total # Method
≤1 - ≥16µg/mL Tested) ≤2 dil. 1 dil. Exact 1 dil. ≥2 dil.
lower lower higher higher
1 9 2 0
1
Proteus spp. 13 (90) 10 (76.92%) b 2 (15.38%)b
(7.69%)
95% CI (49.74 to 91.82%) 95% CI (4.32% to 42.23%)
a
Total number of evaluable results for trending analysis
b
Difference between isolates trending lower and isolates trending higher: 61.54%; 95% CI (23.33% to
80.10%)

A lower MIC result trend was observed in with Proteus spp. compared to the CLSI
broth microdilution reference method and there are concerns for potential very major

Page 10 of 12
 discrepancies. This trending and the potential for occurrence of very major
discrepancies for Gentamicin when testing clinical and challenge isolate results with
the VITEK 2 GN Gentamicin was addressed by adding the following footnote to the
performance table for Gentamicin in the device labeling (Table 127: Performance
Characteristics for Gram-Negative Antimicrobial Susceptibility Testing, VITEK 2
Product Information Manual).

Footnote:
“Of 90 Proteus spp. isolates tested, 13 were evaluable for trending analysis. Based on
this analysis, some VITEK 2 Gentamicin MIC values tended to be at least one
doubling dilution lower when compared to the reference broth microdilution.”

Table 4.2: Trending Analysis of Evaluable Clinical and Challenge

Results for K. pneumoniae
Totala Difference in MIC as Compared to the CLSI Reference
Gentamicin (Total # Method
≤1 - ≥16µg/mL Tested) ≤2 dil. 1 dil. Exact 1 dil. ≥2 dil.
lower lower higher higher
1 2 7 2
1
K. pneumoniae 13 (163) 3 (23.08%)b 9 (69.23%)b
(7.69%)
95% CI (8.18% to 50.26%) 95% CI (42.37% to 87.32%)
a
Total number of evaluable results for trending analysis
b
Difference between isolates trending lower and isolates trending higher: -46.15%; 95% CI (-69.59% to
-7.94%)

A higher MIC result trend was observed with K. pneumoniae compared to the CLSI
broth microdilution reference method and there are concerns for potential major
discrepancies. This trending and the potential for occurrence of major discrepancies
for Gentamicin when testing clinical and challenge isolate results with the VITEK 2
GN Gentamicin was addressed by adding the following footnote to the performance
table for Gentamicin in the device labeling (Table 127: Performance Characteristics
for Gram-Negative Antimicrobial Susceptibility Testing, VITEK 2 Product
Information Manual).

Footnote:
“Of 163 K. pneumoniae isolates tested, 13 were evaluable for trending analysis.
Based on this analysis, some VITEK 2 Gentamicin MIC values tended to be at least
one doubling dilution higher when compared to the reference broth microdilution.”

The analysis of the Pseudomonas aeruginosa MIC data did not demonstrate notable
trending.

b. Matrix comparison:

Not applicable

Page 11 of 12
 3. Clinical studies:

Not applicable

4. Clinical cut-off:

Not applicable

5. Expected values/Reference range:

Table 5: Interpretive Criteria for Gentamicin (FDA Drug Label)

FDA Interpretive Criteria for Gentamicin MIC
Organism (µg/mL)
S I R
Enterobacteriaceae ≤4 8 ≥16
Pseudomonas aeruginosa ≤4 8 ≥16

N. Proposed Labeling:

The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10.

O. Conclusion:

The submitted information in this premarket notification is complete and supports a

substantial equivalence decision.

Page 12 of 12