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®
VIDAS C. difficile GDH (GDH)
VIDAS C. difficile GDH (GDH) is an automated test based on the Enzyme Linked Fluorescent Assay technique (ELFA),
for use on the VIDAS family instruments.
The VIDAS C. difficile GDH (glutamate dehydrogenase) assay is a qualitative test that detects the C. difficile antigen,
glutamate dehydrogenase, as a screen for the presence of C. difficile in fecal specimens from persons suspected of
having C. difficile infection (CDI). With the use of additional tests that detect C. difficile toxins, the test is to be used as an
aid in the diagnosis of C. difficile infection. As with other C. difficile tests, results should be considered in conjunction
with the patient history.

SUMMARY AND EXPLANATION
Normal bacterial flora in the intestine provide an
ecological barrier against significant colonization by
pathogenic organisms. Disruption of this protection by
antibiotic use, may permit overgrowth of endogenous or
nosocomially-acquired pathogens such as Clostridium
difficile.
C. difficile have been found to be the major etiologic agent
of antibiotic-associated pseudomembranous colitis (PMC).
PMC is a clinically defined syndrome, associated with a
recent history of antibiotic use, where
pseudomembranous nodules or plaques form in the distal
and sigmoid colon and rectum (1, 2, 3). If unrecognized or
untreated, the disease can be fatal (4). C. difficile has also
been implicated in antibiotic-associated colitis (AAC) and
antibiotic-associated diarrhea (5).
Nosocomial acquisition of C. difficile is a serious
consideration for some institutions, particularly those with
high inpatient populations, chemotherapy wards, or long-
term patient care (6, 7, 8, 9). Also, infants and cystic
fibrosis (CF) patients have been shown to be
asymptomatic carriers of toxigenic C. difficile with
colonization rates as high as 50% in infants (10) and 32%
in CF patients (11).
Clinical diagnosis of C. difficile-associated disease
(CDAD) is made using a number of criteria, which usually
include the following: patient producing at least 3
unformed stools over a 24-hour period, having received
antibiotic therapy within 8 weeks of onset of diarrhea, no
other obvious cause for diarrhea, and appropriate
response to therapy (12).
Bacteriological diagnosis is based on the detection of
C. difficile toxins B and/or A in stools. Cell culture
cytotoxicity assay and toxigenic culture on selective media
are the two gold standards; however, they are time-
consuming (minimum 24-48 hours) and lack
standardization. Enzyme immunoassays (microplate or
unit tests) are rapid methods which enable the
simultaneous detection of toxins A and B, but which are
insufficiently sensitive to be used as the sole method of
diagnosis (13,14).
To improve the sensitivity and rapidity of diagnosis, a two-
step algorithm strategy is recommended. The first step is
the detection of glutamate dehydrogenase (GDH) in
stools, and the second step is the confirmation of positive
results using a second test for toxin detection (15,16, 17).
To evaluate the test results, the physician must take into
account both the patient history and the limitations of the
method used (18).
PRINCIPLE
The assay principle combines a two-step enzyme
immunoassay sandwich method with a final fluorescent
detection (ELFA).
®
The Solid Phase Receptacle (SPR ) serves as the solid
phase as well as the pipetting device. Reagents for the
assay are ready-to-use and are pre-dispensed in the
sealed reagent strips.
All of the assay steps are performed automatically by the
instrument. The reaction medium is cycled in and out of
the SPR several times. Each step is followed by a wash
cycle which eliminates unbound components:
• Specific binding of GDH present in the sample with
mouse monoclonal anti-GDH antibody coated on the
interior of the SPR.
• Binding between GDH and mouse monoclonal anti-GDH
antibody conjugated with alkaline phosphatase (ALP).
• Detection: alkaline phosphatase catalyzes the hydrolysis
of the substrate (4-Methyl-umbelliferyl phosphate) into a
fluorescent product (4-Methyl-umbelliferone) the
fluorescence of which is measured at 450 nm.
The intensity of the fluorescence increases according to
the quantity of GDH in the sample.
When the VIDAS C. difficile GDH test is completed, the
results are analyzed automatically by the instrument, a
test value is generated, and a result is printed for each
sample.

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VIDAS C. difficile GDH (GDH) 9302513 D - en - 2015/01

CONTENT OF THE KIT (60 TESTS) – RECONSTITUTION OF REAGENTS:

60 GDH Strips STR Ready-to-use.
®
60 GDH SPRs SPR Ready-to-use.
(2 x 30) Interior of SPRs coated with mouse monoclonal anti-C. difficile GDH
antibody.
Standard GDH S1 Reconstitute with 1 mL distilled water. Wait for 5 to 10 minutes and then
2 x 1 mL (lyophilized) mix. After reconstitution, stable 2 months at 2-8°C.
Dilution of recombinant GDH from C. difficile in buffer 0.1 mol/L (pH 7.2)
+ Protein stabilizer + preservatives.
MLE data indicate the confidence interval in "Relative Fluorescence
Value (RFV)" ("Standard (S1) RFV Range").
Positive Control GDH C1 Reconstitute with 2 mL distilled water. Wait for 5 to 10 minutes and then
2 x 2 mL (lyophilized) mix. After reconstitution, stable 2 months at 2-8°C.
Dilution of recombinant GDH from C. difficile in buffer 0.1 mol/L (pH 7.2)
+ Protein stabilizer + preservatives.
MLE data indicate the Test Value (TV) range: Control C1 (+) Test Value
Range.
Negative Control GDH C2 Buffer 0.1 mol/L (pH 7.2) + Protein stabilizer + preservatives.
1 x 5.6 mL (liquid) MLE data indicate the Test Value (TV) range: Control C2 (-) Test Value
Range.
Pretreatment Reagent R1 Ready-to-use. Buffer 0.1mol/L (pH 7.2) + foetal calf serum + detergent +
1 x 61 mL (liquid)
C. difficile preservatives.
Specifications for the factory master data required to calibrate the test:
• MLE data (Master Lot Entry) provided in the kit,
or
• MLE bar codes printed on the box label.
1 Package Insert provided in the kit or downloadable from www.biomerieux.com/techlib.

The SPR: The Reagent Strip:

The interior of the SPR is coated during production with The strip consists of 10 wells covered with a labeled, foil
mouse monoclonal anti- C. difficile GDH antibody. Each seal. The label contains a bar code which includes the
SPR is identified by the code "GDH" code. Only remove assay code, kit lot number and expiration date. The foil of
the required number of SPRs from the pouch and the first well is perforated to facilitate the introduction of
carefully reseal the pouch after opening. the sample. The last well of each strip is a cuvette in
which the fluorometric reading is performed. The eight
wells in the center section of the strip contain the various
reagents required for the assay.
Description of the GDH strip:
Wells Reagents
1 Sample well.
2-3-4 Wash solution: Buffer 0.2 mol/L (pH 7.8) + detergent + preservatives (600 µL).
5 Conjugate: mouse monoclonal anti-C. difficile GDH antibody labeled with ALP +
Protein stabilizer + preservative (400 µL).
6-7-8-9 Wash solution: Buffer 0.2 mol/L (pH 7.8) + detergent + preservatives (600 µL).
10 Reading cuvette with substrate: 4-Methyl-umbelliferyl phosphate (0.6 mmol/L) +
diethanolamine DEA* (0.62 mol/L or 6.6%) pH 9.2 + sodium azide 1g/L (300 µL).

* Signal Word: DANGER

Hazard statement
H318 : Causes serious eye damage.

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VIDAS C. difficile GDH (GDH)
Precautionary statement
P280 :Wear protective gloves/protective clothing/eye protection/face protection.
P305 + P351 + P338: IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present
and easy to do. Continue rinsing.
For further information, refer to the Material Safety Data Sheet.
MATERIALS AND DISPOSABLES REQUIRED BUT
NOT PROVIDED
- Pipette with disposable tip to dispense 200 µL,
1000 µL and 2000 µL.
- Powderless disposable gloves.
- Centrifuge (≥ 3000 g). Must be used according to the
manufacturer’s recommendations.
- Polypropylene or other appropriate centrifuge tubes for
specimen dilution and centrifugation (with at least
1.5 mL capacity).
- Sample transfer pipets used to measure the appropriate
quantity of stool sample for processing.
- Applicator stick or loop.
- For other specific materials and disposables, please
refer to the Instrument User’s Manual.
- Instrument of the VIDAS family.
WARNINGS AND PRECAUTIONS
• For in vitro diagnostic use only.
• For professional use only.
• This kit contains products of animal origin. Certified
knowledge of the origin and/or sanitary state of the
animals does not totally guarantee the absence of
transmissible pathogenic agents. It is therefore
recommended that these products be treated as
potentially infectious and handled observing the
usual safety precautions (do not ingest or inhale).
• Kit reagents contain sodium azide which can react with
lead or copper plumbing to form explosive metal azides.
If any liquid containing sodium azide is disposed of in
the plumbing system, drains should be flushed with
water to avoid build-up.
• Do not use reagents after the expiration date indicated
on the label.
• Do not mix reagents or disposables from different lots.
• Do not use the SPRs if the pouch is pierced or if the dot
sealing a SPR has come unstuck.
• Do not use visibly deteriorated STRs (damaged foil or
plastic).
• Use powderless gloves, as powder has been reported
to cause false results for certain enzyme immunoassay
tests (19).
• Use calibrated pipettes to process standard, positive
and negative controls.
• The optical cuvette with substrate (well 10) contains an
irritant agent (diethanolamine 6.6%). Refer to the hazard
statements "H" and the precautionary statements "P"
above.
• Spills should be wiped up thoroughly after treatment
with liquid detergent or a solution of household bleach
containing at least 0.5% sodium hypochlorite. See the
User’s Manual for cleaning spills on or in the instrument.
Do not autoclave solutions containing bleach.
• The instrument should be regularly cleaned and
decontaminated (see the User’s Manual).
STORAGE CONDITIONS
• Store the VIDAS C. difficile GDH kit (SPRs, strips,
standard, controls and pretreatment reagent) at
2-8°C
• Do not freeze reagents.
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• Store all unused reagents at 2-8°C.
• After opening the kit, check that the SPR pouch is
correctly sealed and undamaged. If not, do not use the
SPRs.
• Carefully reseal the pouch with the desiccant inside
after use to maintain stability of the SPRs and return
the complete kit to 2-8°C.
• If stored according to the recommended conditions, all
components are stable until the expiration date indicated
on the label.
SPECIMENS
Specimen type, collection and storage:
Stool specimens should be collected and transported
according to standard laboratory procedures.
Stool specimens may be stored at 2-8°C for 3 days (from
time of collection) prior to processing.
If longer storage is required, freeze the specimens at
-70°C+/- 10°C. Avoid repeated freezing and thawing
cycles.
A rectal swab will not provide enough specimen for the
test and therefore is unacceptable. Do not use containers
which may contain detergents, preservatives or media
that may interfere with the VIDAS C. difficile GDH assay
results.
Standard, control, and specimen preparation for the
VIDAS C. difficile GDH test:
Specimens:
Important: it is critical that stool specimens be thoroughly
mixed before sample processing is begun. Lack of
homogeneity in a stool sample may lead to incorrect
results. Thorough mixing of stool specimens is essential
to avoid this problem. The diluted sample must be
homogeneous after mixing. This will help ensure valid
results.
For this sample preparation step, it is essential to
check the amount of deposited sample regardless of
the consistency of the stools (liquid, semi-solid or
solid).
Liquid stools:
1. Homogenize by drawing the stool sample in and out of
the transfer pipet.
Using the same transfer pipet, dispense 200 µL of
liquid stool in a clean centrifuge tube.
2. Using a pipette with disposable tip, add 1000 µL of
pretreatment reagent (R1 C. difficile) to the centrifuge
tube.
Note: Pretreatment reagents other than VIDAS
C. difficile GDH must not be used with the VIDAS
C. difficile GDH assay.
3. Homogenize again by drawing in and out of the transfer
pipet.
Then mix using a vortex-type mixer until a
homogeneous medium is obtained.
It is essential that all portions of the sample be
uniformly mixed with pretreatment reagent
(R1 C. difficile).
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VIDAS C. difficile GDH (GDH)
4. Centrifuge for 10 minutes at 2-25°C at a minimum of
3000 g.
5. Using a pipette with disposable tip, collect 300 µL of
supernatant to perform the VIDAS C. difficile GDH test.
Note: If particulate matter or lipid material remain at the
surface of the supernatant after centrifugation, collect the
sample below the surface layer.
Semi-solid and solid stools:
1. To ensure the precise transfer of 200 mg of semi-solid
or solid stool, dispense 200 µL of distilled water into a
clean centrifuge tube using a pipette with disposable
tip; this tube will be used as a “gauged tube”.
2. Using a wooden applicator stick, transfer a quantity of
stool (200 mg) equivalent to the volume in place in the
gauged tube, into a new clean centrifuge tube
identified as the sample tube (see hereafter). Deposit
the stool sample at the bottom of the centrifuge
tube so that the quantity of stool is equivalent to the
volume of water in the gauged tube (200 µL).
Equivalent
volume
“Gauged tube”: Sample tube:
200 µL distilled water 200 mg stool
sample
Note: Discard the gauged tube after transferring each of
the solid and semi-solid stool specimens.
3. Add 1000 µL of pretreatment reagent (R1 C. difficile)
to the sample tube using a pipette with disposable tip.
Note: Pretreatment reagents other than VIDAS
C. difficile GDH must not be used with the VIDAS
C. difficile GDH assay.
4. Emulsify the stool sample using the applicator stick,
and then mix using a vortex-type mixer until the
sample appears homogeneous. It is essential that all
portions of the sample be homogeneously mixed
with pretreatment reagent (R1 C. difficile).
5. Centrifuge for 10 minutes at 2–25°C at a minimum of
3000 g.
6. Using a pipette with disposable tip, collect 300 µL of
supernatant to perform the VIDAS C. difficile GDH
test.
Note:
- If particulate matter or lipid material remain at the
surface of the supernatant after centrifugation, collect
the sample below the surface layer;
- In cases where semi-solid stools consist more of liquid
than of solid, the sample preparation procedure may
be performed using the instructions provided for liquid
stools (see section “Specimens: Liquid Stools”).
Standard and Controls:
The standard and the positive and negative controls
must be processed as follows :
1. Reconstitute one vial of standard with 1 mL distilled
water. Wait for 5 to 10 minutes and then mix. After
reconstitution, the vials are stable for 2 months at
2-8°C.
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2. Reconstitute one vial of positive control with 2 mL
distilled water. Wait for 5 to 10 minutes and then mix.
After reconstitution, the vials are stable for
2 months at 2-8°C.
3. Mix reconstituted standard, reconstituted positive
control and negative control vigorously using a
vortex-type mixer
4. Collect 200 µL of reconstituted standard,
reconstituted positive control and negative control
using a pipette with disposable tip and dispense into
a clean tube.
5. Add 1000 µL of pretreatment reagent (R1 C. difficile)
to the tube using a pipette with disposable tip.
6. Mix using a vortex-type mixer.
7. Using a pipette with disposable tip, collect 300 µL to
perform the VIDAS C. difficile GDH test.
Storage of specimen supernatant and processed
standard and controls:
Specimen supernatants, processed standard and controls
may be stored up to 8 hours at 18-25°C or 48 hours at
2-8°C before being tested with the VIDAS C. difficile GDH
assay.
INSTRUCTION FOR USE
For complete instructions, see the User’s Manual.
Reading VIDAS® Protocole Test Change (PTC)
protocol data and MLE data
When using the assay for the first time:
With the external instrument barcode reader,
1. Scan the PTC barcode(s) at the end of the package
insert. or downloadable from
www.biomerieux.com/techlib. This reading allows
VIDAS® PTC protocol data to be transferred to the
instrument software for its update.
2. Scan the MLE data on the box label.
Note: If the MLE data have been read before the
®
VIDAS PTC protocol, read the MLE data again.
When opening a new lot of reagents:
Enter the specifications (or factory master data) into the
instrument using the master lot entry (MLE) data.
If this operation is not performed before initiating the tests,
the instrument will not be able to print results.
Note: the master lot data need only be entered once
for each lot.
It is possible to enter MLE data manually or
automatically depending on the instrument (refer to the
User’s Manual).
Calibration
The calibration, using the standard provided in the kit,
must be performed each time a new lot of reagents is
opened, after the master lot data have been entered.
Calibration should then be performed every 28 days. This
operation provides instrument-specific calibration
information and compensates for possible minor
variations in assay signal throughout the shelf-life of the
kit.
The standard, identified by S1, and processed as defined
above (standard, controls and specimen preparation)
must be tested in duplicate (see User’s Manual). The
standard value must be within the set RFV "Relative
Fluorescence Value" range. If this is not the case,
recalibrate.
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VIDAS C. difficile GDH (GDH)
PROCEDURE
1. Only remove the required reagents from the
refrigerator. They can be used immediately.
2. Use one "GDH" strip and one "GDH" SPR from the kit
for each sample supernatant, processed control, and
processed standard to be tested. Make sure the
storage pouch has been carefully resealed after
the required SPRs have been removed.
3. The test is identified by the "GDH" code on the
instrument. The processed standard must be identified
by "S1", and tested in duplicate.
4. For this test, the processed standard, controls and
sample supernatant test portion is precisely
300 µL (20)
5. Insert the "GDH" SPRs and "GDH" strips into the
instrument. Check to make sure the color labels with
the assay code on the SPRs and the Reagent Strips
match.
6. Initiate the assay as directed in the User’s Manual. All
the assay steps are performed automatically by the
instrument.
7. Reclose the vials and return them to the
recommended temperature after pipetting.
8. The assay will be completed within approximately
50 minutes. After the assay is completed, remove the
SPRs and strips from the instrument.
9. Dispose of the used SPRs and strips into an
appropriate receptacle.
RESULTS
Fluorescence is measured twice in the reagent strip’s
reading cuvette for each sample tested. The first reading
is a background reading of the substrate cuvette before
the SPR is introduced into the substrate. The second
reading is taken after incubating the substrate with the
enzyme remaining on the interior of the SPR. The RFV
(Relative Fluorescence Value) is calculated by subtracting
the background reading from the final result. This
calculation appears on the result sheet.
The test value is calculated for each sample by the
instrument as follows:
Test Value = patient RFV / standard RFV
This test value as well as the interpretation appears on
the result sheet. The clinical cut-off was established at a
Test Value of 0.10. Interpretation according to the Test
Value is as follows:
Test Value Result
< 0.10 Negative
≥ 0.10 Positive
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QUALITY CONTROL
One positive control and one negative control are included
in each VIDAS C. difficile GDH kit.
These controls must be performed each time a new lot of
reagents is opened to ensure that reagent performance
has not been altered. Each calibration must be checked
using these controls. The instrument will only be able to
check the control values if they are identified as C1 and
C2.
Results cannot be validated if the control values deviate
from the expected values.
Note
It is the responsibility of the user to perform Quality
Control in accordance with any local applicable
regulations.
LIMITATIONS OF THE METHOD
1. Due to sample heterogeneity, thorough mixing of stool
specimens is essential to avoid discrepant results.
Samples giving results contradictory to the clinical
information available should be retested using a fresh
specimen.
2. A negative VIDAS C. difficile GDH result alone may
not rule out the possibility of C difficile-associated
colitis or diarrhea. It could be the result of inadequate
sampling, handling or sample storage. Always
evaluate VIDAS C. difficile GDH assay results along
with clinical signs and patient history when diagnosing
C. difficile-related disease.
3. A positive VIDAS C. difficile GDH result alone should
not be used to diagnose C. difficile-associated colitis
or diarrhea. VIDAS C. difficile GDH assay results
should always be evaluated along with clinical signs
and patient history when diagnosing C. difficile-related
disease.
4. Guidelines define the best strategy for C. difficile
testing as testing performed only on diarrheal
(unformed) stool, unless ileus due to C. difficile is
suspected. The proper specimen for the diagnosis of
C. difficile infection is a watery, loose or unformed
stool. Evaluating a formed stool can impact the
specificity of diagnosis of CDAD (12, 15, 16 and 17).
EPIDEMIOLOGICAL DATA
In a European study (21) conducted by the ESGCD
(European Study Group on Clostridium difficile) the
average frequency of stools with positive toxin results
collected among 136 hospitals was 9.5%. In North
America, from a Canadian survey (22) including 380
hospitals, average test positivity rates varied from 13.2 to
17.2% depending on the hospital size (<300 to >500
beds), with an average (Clostridium difficile-Associated
Diseases) incidence between 23.5 and 40.3 cases per
100 000 patient days. In the Society for Healthcare
Epidemiology of America (SHEA) position paper, the
reported rate was 17 to 60 cases per 100 000 bed days
(23).
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VIDAS C. difficile GDH (GDH) 9302513 D - en - 2015/01

CLINICAL PERFORMANCE

Assay sensitivity and specificity

1904 stool samples collected from patients suspected of having C. difficile infection (CDI), were tested at three sites
(Europe and USA). The age groups of the patients range from 1 year to 100 years. A single replicate of each sample was
tested using VIDAS C. difficile GDH on a VIDAS instrument. A bacterial culture test was performed for each sample on a
®
CCFA medium and a chromogenic medium (chromID C. difficile ref 43871). The typical colonies were confirmed using
biochemical or immunological methods.
Data from the study are summarized in the following tables:
Performance of the VIDAS C. difficile GDH assay versus bacterial culture CCFA
CCFA bacterial culture test
Positive Negative Total
VIDAS Positive 293 160* 453
C. difficile GDH
Negative 13** 1438 1451
Total 306 1598 1904

Performance % [95%CI]
Sensitivity 95.8% [92.8 – 97.7]%
Specificity 90.0% [88.4 - 91.4]%
Negative Predictive Value 99.1% [98.5 - 99.5]%
(NPV)
* 160 samples were found positive with the VIDAS C. difficile GDH assay and negative with the CCFA bacterial culture
test, 75 of which were found positive and 85 negative with the commercially available C. difficile GDH assay.
** 13 samples were found negative with the VIDAS C. difficile GDH assay and positive with the CCFA bacterial culture
test, 9 of which were found negative and 4 positive with the commercially available C. difficile GDH assay.
Performance of the VIDAS C. difficile GDH assay versus chrom® ID C. difficile
chromID® C. difficile bacterial
culture test
Positive Negative Total
VIDAS Positive 325 128* 453
C. difficile
25** 1426 1451
GDH Negative

Total 350 1554 1904

Performance % [95%CI]
Sensitivity 92.9% [89.6 - 95.3]%
Specificity 91.8% [90.3 - 93.1]%
Negative Predictive 98.3% [97.5 - 98.9]%
Value (NPV)
* 128 samples were found positive with the VIDAS C. difficile GDH assay and negative with the chromID® C. difficile
bacterial culture test, 49 of which were found positive and 79 negative with the commercially available C. difficile GDH
assay.
®
** 25 samples were found negative with the VIDAS C. difficile GDH assay and positive with the chromID C. difficile
bacterial culture test, 18 of which were found negative and 7 positive with the commercially available C. difficile GDH
assay.

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Sensitivity & Specificity performances versus CCFA medium by age group

N N
Age Group Sensitivity [CI95] % Specificity [CI95] %
(Positive samples) (Negative samples)
< 2 years 1 100.0% [2.5 - 100]% 2 100.0% [15.8 - 100]%
2-12 years 22 100.0% [84.6 - 100]% 47 85.1% [71.7 - 93.8]%
13-21 years 13 100.0% [75.3 - 100]% 45 88.9% [75.9 - 96.3]%
22-59 years 125 97.6% [93.1 - 99.5]% 632 88.9% [86.2 - 91.3]%
≥ 60 years 145 93.1% [87.7 - 96.6]% 872 91.1% [89.0 - 92.9]%

Method comparison with a commercially available C. difficile GDH assay

1904 stool samples collected from patients suspected of having C. difficile infection (CDI) were tested at 3 sites (Europe
and USA). A single replicate of each sample was tested using VIDAS C. difficile GDH on a VIDAS instrument and a
commercially available C. difficile GDH assay. Data from the study are summarized in the following table:
Comparison between the VIDAS C. difficile GDH assay and the commercially available C. difficile GDH assay
Commercially available
C. difficile GDH assay (all sites)
Positive Negative Total
VIDAS 367 86 453
Positive
C. difficile
10 1441 1451
GDH Negative

Total 377 1527 1904

Performance % [95%CI]
Positive percent agreement 97.3% [95.2 - 98.7]%

Negative percent agreement 94.4% [93.1 - 95.5]%

Overall percent agreement 95.0% [93.9 – 95.9]%

ANALYTICAL PERFORMANCES
Precision
The within-laboratory precision was estimated at one site based on the recommendations of the CLSI® EP5-A2
Three human samples, including 2 close to the clinical cut-off (1 high negative and 1 low positive) were tested in triplicate
in 2 runs per day with 2 different operators, with 2 reagent lots for a total of 12 testing days (6 days of test per lot) on 1
VIDAS instrument (N=72 test values for each sample). Two calibrations were used for each reagent lot (3 days of test
per calibration and lot) over the whole period of the study. Data from the study are summarized in the following table:

Total within-laboratory
precision (total within-
Repeatability
instrument, between-lot,
Sample N Mean test value between-calibration)
Standard Standard
CV (%) CV (%)
deviation deviation
Sample 1
72 0.07 0.00 6.0 0.01 14.1
High negative
Sample 2
72 0.12 0.01 5.2 0.01 11.9
Low positive
Sample 3
Moderate 72 0.27 0.02 5.7 0.03 11.2
positive

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The total between-instrument between-lot reproducibility was estimated at three sites based on the recommendations of
®
the CLSI EP5-A2.
Three human samples, including 2 close to the clinical cut-off (1 high negative and 1 low positive), were tested in
triplicate in 2 runs per day with 2 different operators, using 2 reagent lots for a total of 6 testing days (3 days of test for
each lot) on 3 VIDAS instruments at 3 different sites (N=108 test values for each sample). One calibration was used for
each reagent lot over the whole period of the study. Data from the study are summarized in the following table:

Reproducibility
(total between sample
Sample N Mean test value preparation/operator/run/day/lot/
instrument)
Standard deviation CV (%)

Sample 1
108 0.06 0.01 19.1
High negative
Sample 2
108 0.12 0.02 12.9
Low positive
Sample 3
Moderate 108 0.26 0.03 13.0
positive

Analytical reactivity
The VIDAS C. difficile GDH assay was also evaluated using several strains of C. difficile. Strains were grown on
Columbia agar + 5% sheep blood (bioMérieux ref. 43041) and tested, using the VIDAS C. difficile GDH assay, at a
8
concentration of 9x10 CFU/mL (3 McFarland).
The VIDAS C. difficile GDH detects the following C.difficile strains:
Toxinogenic C.difficile: ATCC 43255TM; ATCC 9689TM; ATCC 700792TM; ATCC17858TM; ATCC BAA–1805TM;
ATCC BAA-1382TM; ATCC 51695TM; ATCC 43600TM; ATCC 43599TM; ATCC 43596TM; ATCC 43594TM; ATCC
17857TM; ATCC 43598TM; CCUG 20309.
Non toxinogic C.difficile strains: ATCC 700057TM; ATCC 43593TM; X1a IS58; X1b R1 1402; ATCC 43601TM
8
(3x10 CFU/mL).
The Cardiff ECDC collection including the following ribotypes:
001(7 strains); 002; 003; 012; 014; 015; 017; 020; 023; 027; 029; 046; 053; 056; 070; 075; 077; 078; 081; 087; 095; 106;
126; 131; VPI 10463; 005; 010; 045; 048; 156; 174.
The bioMerieux collection including the following ribotypes:
001 (6 strains); 002 (9 strains); 005 (2 strains); 010 (1 strain); 012 (4 strains); 014 (10 strains) ;015 (1 strain); 017 (20
strains); 020 (5 strains); 023 (1 strain); 027 (24 strains); 047 (1 strain); 050 (1 strain); 053 (4 strains); 054 (2 strains); 056
(2 strains);057 (1 strain); 058 (1 strain); 075 (1 strain); 078 (3 strains); 096 (1 strain); 097 (1 strain); 103 (2 strains); 106
(16 strains); 110 (2 strains); 118 (1 strain); 153 (1 strain); 177 (1 strain).
Analytical sensitivity
Sensitivity at clinical cut-off
The sensitivity at the clinical cut-off (test value = 0.10) was evaluated using a range of dilutions of purified native C.difficile
GDH and recombinant C. difficile GDH in a pool of C. difficile-negative stool samples.
The VIDAS C. difficile GDH assay detects purified native GDH at a level of 2.13 ng/mL and recombinant GDH at a
level of 0.70 ng/mL at the clinical cut-off (test value = 0.10).
Limit of detection
The limit of detection was evaluated using a range of dilutions of purified native C.difficile GDH and recombinant
C. difficile GDH in a pool of C. difficile-negative stool samples based on the recommendations of the CLSI EP17-A.
The limit of detection of the VIDAS C. difficile GDH assay (95% detection rate for positive samples) is 3.00 ng/mL for
purified native GDH and 0.75 ng/mL for recombinant GDH.
Hook effect
No hook effect was observed up to purified native GDH concentrations of 2 µg/mL, and recombinant GDH
concentrations of 600 ng /mL.

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Interferences

Study of drug interferences and other potentially interfering substances

Potential interferences by commonly used drugs and other substances was determined based on the recommendations
®
of the CLSI EP7-A2, at 2 levels of GDH (a low positive close to the clinical cut-off and a high positive).
No significant interference No significant
observed up to a interference observed
Tested compound Tested compound
concentration of: up to concentrations
of:
Hemoglobin 3.2 mg/mL Hydrocortisone 0.39 mg/mL
Lipids 20 mg/mL Aluminium hydroxide 15.3 mg/mL
Mucine 3.33 mg/mL Magnesium hydroxide 6.2 mg/mL
Amoxicillin 196.49 µmol/L Lidocaine 0.12 mg/mL
Bismuth salicylate 8.2 mg/mL Loperamide 0.08 mg/mL
Calcium carbonate 13.06 mg/mL Mesalazine 19.2 mg/mL
Ceftriaxone 1.46 mmol/L Metronidazole 2 mg/mL
Benzalkonium chloride 2 µg/mL Naproxen 2170 µmol/L
Ciprofloxacin 30.2 µmol/L Nystatin 600 UI/mL
Erythromycin 81.6 µmol/L Phenylephrine 0.16 mg/mL
Ethanol 86.8 mmol/L Sennosides 0.24 mg/mL
Fidaxomicin 4 mg/mL Tergitol 0.125 v/v
Gentamicin 21 µmol/L Tetracycline 34 µmol/L
Mineral oil 0.27 v/v Vancomycin 5 mg/mL

Cross-reactivity and interference:

To test for cross-reactivity, each micro-organism was diluted in a pool of C. difficile-negative stool samples, pretreated
and a single replicate was tested using the VIDAS C. difficile GDH assay.
To test for interference, each micro-organism was diluted in a pool of C. difficile-positive stool samples, pretreated and a
single replicate was tested using the VIDAS C. difficile GDH assay. The micro-organisms were tested at a concentration
8 5
of 3x10 CFU/mL (1 McFarland) for bacteria and 1x10 PFU/mL for viruses.
None of the following micro-organisms, present in the stool samples, reacted with the VIDAS C. difficile GDH assay:
Abiotrophia defectiva, Acinetobacter baumannii, Acinetobacter lwoffii, Aeromonas hydrophila ssp hydrophila, Alcaligenes
faecalis ssp faecalis, Anaerococcus tetradius, Bacillus cereus, Bacteroides caccae, Bacteroides merdae, Bacteroides
stercoris, Bifidobacterium adolescentis, Bifidobacterium longum, Campylobacter jejuni ssp jejuni, Candida albicans,
Candida catenulata, Cedecea davisae, Chlamydia trachomatis, Citrobacter amalonaticus, Citrobacter freundii,
Citrobacter koseri, Citrobacter sedlakii, Clostridiuim nexile, Clostridium beijerinckii, Clostridium bifermentans, Clostridium
bolteae, Clostridium butyricum, Clostridium chauvoei, Clostridium fallax, Clostridium haemolyticum, Clostridium
histolyticum, Clostridium innocuum, Clostridium novyi, Clostridium paraputrificum, Clostridium perfringens, Clostridium
ramosum, Clostridium scindens, Clostridium septicum, Clostridium sordellii, Clostridium sphenoides, Clostridium
spiroforme, Clostridium sporogenes, Clostridium symbosum, Clostridium tertium, Clostridium tetani, Collinsella
aerofaciens, Corynebacterium genitalium, Desulfovibrio piger, Edwardsiella tarda, Eggerthella lenta, Enterobacter
aerogenes, Enterobacter cloacae, Enterococcus casseliflavus, Enterococcus cecorum, Enterococcus dispar,
Enterococcus faecalis, Enterococcus faecium, Enterococcus gallinarum, Enterococcus hirae, Enterococcus raffinosus,
Escherichia coli, Escherichia fergusonii, Escherichia hermannii, Flavonifractor plautii, Fusobacterium varium, Gardnerella
vaginalis, Gemella morbillorum, Hafnia alvei, Helicobacter fenneliae, Helicobacter pylori, Klebsiella oxytoca, Klebsiella
pneumoniae ssp pneumoniae, Lactobacillus acidophilus, Lactobacillus reuteri, Lactococcus lactis ssp lactis, Leminorella
grimontii, Listeria grayi, Listeria innocua, Listeria monocytogenes, Peptoniphilus asaccharolyticus, Peptostreptococcus
anaerobius, Plesiomonas shigelloides, Porphyromonas asaccharolytica, Prevotella melaninogenica, Proteus mirabilis,
Proteus penneri, Providencia alcalifaciens, Providencia rettgeri, Providencia stuartii, Pseudomonas aeruginosa,
Pseudomonas putida, Salmonella enterica ssp arizonae, Salmonella ser.Choleraesuis, Salmonella ser.Typhimurium
Serratia liquefaciens, Serratia marcescens, Shigella boydii, Shigella dysenteriae, Shigella sonnei, Staphylococcus
aureus ssp aureus, Staphylococcus epidermidis, Stenotrophomonas maltophilia, Streptococcus agalactiae,
Streptococcus dysgalactiae ssp dysgalactiae,Streptococcus intermedius, Streptococcus uberis, Trabulsiella guamensis,
Veillonella parvula, Vibrio cholerae, Vibrio parahaemolyticus,Yersinia bercovieri, Yersinia rohdei, Adenovirus 40 et 41,
Rotavirus RF, Norovirus, Enterovirus 70, Echovirus 12, Coxsackie virus, Cytomegalovirus AD169.

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WASTE DISPOSAL 16. COHEN SH, et al. 2010. Clinical practice guidelines for
Clostridium difficile infection in adults: 2010 update by the
Dispose of used or unused reagents as well as any other Society for Healthcare Epidemiology of America (SHEA)
contaminated disposable materials following procedures and the Infectious Diseases Society of America (IDSA).
for infectious or potentially infectious products. Infect Control Hosp Epidemiol;31(5):431–455.
It is the responsibility of each laboratory to handle waste 17. CHENG A.C. et al. 2011, Australasian Society for
and effluents produced according to their nature and Infectious Diseases guidelines for the diagnosis and
degree of hazardousness and to treat and dispose of treatment of Clostridium difficile infection. Medical Journal
them (or have them treated and disposed of) in of Australia 194 (7) 353–358.
accordance with any applicable regulations. 18. RUPNIK M., WILCOX MH. and GERDING DN., 2009.
Clostridium difficile infection: new developments in
epidemiology and pathogenesis, Nat Rev Microbiol 7,
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WARRANTY
bioMérieux disclaims all warranties, express or implied, including any implied warranties of MERCHANTABILITY AND
FITNESS FOR A PARTICULAR USE. bioMérieux shall not be liable for any incidental or consequential damages. IN NO
EVENT SHALL BIOMERIEUX’S LIABLITY TO CUSTOMER UNDER ANY CLAIM EXCEED A REFUND OF THE
AMOUNT PAID TO BIOMERIEUX FOR THE PRODUCT OR SERVICE WHICH IS THE SUBJECT OF THE CLAIM.

REVISION HISTORY
Change type categories :
N/A Not applicable (First publication)
Correction Correction of documentation anomalies
Technical change Addition, revision and/or removal of information related to the product
Administrative Implementation of non-technical changes noticeable to the user
Note: Minor typographical, grammar, and formatting changes are not included in the
revision history.
Release
Part Number Change Type Change Summary
date
INDEX OF SYMBOLS
Administrative
REVISION HISTORY
2015/01 9302513D CONTENT OF THE KIT (60 TESTS) –
Technical RECONSTITUTION OF REAGENTS
WARNINGS AND PRECAUTIONS

BIOMERIEUX, the blue logo, chromID, SPR and VIDAS are used, pending and/or registered trademarks belonging to bioMérieux or one
of its subsidiaries, or one of its companies.
CLSI is a trademark belonging to Clinical and Laboratory Standards Institute Inc.
The ATCC trademark and trade name and any and all ATCC catalog numbers are trademarks of the American Type Culture Collection.
Any other name or trademark is the property of its respective owner.

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