REF 43 871 15010 B - en - 2011/03

chromID™ C. difficile agar (CDIF)

Chromogenic medium for the detection and identification of Clostridium difficile
.

SUMMARY AND EXPLANATION WARNINGS AND PRECAUTIONS

chromID C. difficile agar is a selective chromogenic • For in vitro diagnostic use only.
medium for the detection and identification of Clostridium • For professional use only.
difficile in human specimens (stools of symptomatic • This kit contains products of animal origin. Certified
patients) (1, 2). knowledge of the origin and/or sanitary state of the
The medium contributes to the diagnosis and animals does not totally guarantee the absence of
epidemiological monitoring of Clostridium difficile transmissible pathogenic agents. It is therefore
infections (3). recommended that these products be treated as
Clostridium difficile is a causative agent of potentially infectious and handled observing the usual
pseudomembranous colitis and more generally of safety precautions (do not ingest or inhale).
nosocomial or antibiotic-associated diarrhea (4, 5, 6, 7, 8). • All specimens, microbial cultures and inoculated
products should be considered infectious and handled
PRINCIPLE appropriately. Aseptic technique and usual precautions
chromID C. difficile agar consists of a rich nutritive base for handling the bacterial group studied should be
®
observed throughout this procedure. Refer to “CLSI
combining different peptones and taurocholate which
M29-A, Protection of Laboratory Workers from
favors the germination of spores (9).
Occupationally Acquired Infections; Approved Guideline
It contains a chromogenic substrate (10) (patent pending)
– Current Revision". For additional information on
and a mixture of antibiotics which enable:
handling precautions, refer to "Biosafety in
• the detection and identification of ß-glucosidase- Microbiological and Biomedical Laboratories – CDC/NIH
producing Clostridium difficile strains based on the – Latest edition", or the current regulations in the
typical grey to black color of colonies. country of use.
• the inhibition of most Gram-positive and Gram-negative • Culture media should not be used as manufacturing
bacteria, yeasts and molds. material or components.
• Do not use reagents after the expiry date.
CONTENT OF THE KIT • Do not use reagents if the packaging is damaged.
• The use of this medium may be difficult for people who
Ready-to-use medium have problems recognizing colors.
REF 43 871 Pack of 2 x10 plates (90 mm) • Do not use contaminated plates or plates that exude
moisture.
CDIF *
• Some precipitates and/or light halos may be observed in
* printed on each plate the agar but these do not affect the performance of the
medium.
COMPOSITION • The reading and interpretation of the medium should be
performed using isolated colonies.
Theoretical formula.
• Interpretation of test results should be made taking into
This medium can be adjusted and/or supplemented according to the
performance criteria required:
consideration colonial and microscopic morphology and,
Meat peptone (porcine).....................................................................8.0 g
if necessary, the results of any other tests performed.
Taurocholate (bovine)..........................................................................1 g • Performance data presented were obtained using the
Yeast extract.....................................................................................3.5 g procedure indicated in this package insert. Any change
Sodium chloride................................................................................6.0 g or modification in the procedure may affect the results.
Selective mixture ............................................................................0.27 g
Chromogenic mixture........................................................................0.3 g
Agar................................................................................................13.0 g STORAGE CONDITIONS
Purified water........................................................................................ 1 l
• Store the plates at 2-8°C in their box until the expiry
pH 7.3 date.
• If not in the box, plates can be stored in the cellophane
MATERIAL REQUIRED BUT NOT PROVIDED sachet for 2 weeks at 2-8°C in the dark.
• Bacteriology incubator. SPECIMENS
• GENbox anaer (Ref. 96124) with jar.
• GENbag anaer (Ref. 45534). The medium should be directly inoculated with liquid
or diarrheal stools or soft stools.
• Thermoregulated chambers with a controlled The specimen can be treated or not with ethanol (11).
atmosphere. Good laboratory practices for collection and transport
should be respected and adapted to anaerobic bacteria.

bioMérieux SA English - 1
chromID™ C. difficile agar (CDIF) 15010 B - en - 2011/03

INSTRUCTIONS FOR USE PERFORMANCE

1. Allow plates to come to room temperature. A study was performed at 2 sites (Spain and Germany)
2. Inoculate the specimen immediately after reception in using clinical specimens.
the laboratory. 737 specimens (liquid diarrheal stools or soft stools) were
TM
inoculated on chromID C. difficile agar and on a
3. Put the plate in a suitable atmosphere (anaerobiosis) if
conventional medium for which reading is recommended
necessary using a controlled atmosphere generator.
after 48 hours.
4. Incubate the inverted plates at 37°C.
The fecal specimens were inoculated either directly
The cultures are examined after 24 hours of
(n=474), or after treatment with ethanol (n=263).
incubation.
Specimens inoculated directly (without ethanol treatment):
READING AND INTERPRETATION Out of the 474 specimens tested, 78 were confirmed to be
• After incubation, observe the bacterial growth and the positive for C. difficile by PCR.
presence of typical colonies: Clostridium difficile
colonies are grey to black with an irregular or smooth Recovery %
Sensitivity (%)
border. (*)
• Appropriate tests should be performed on the specimen chromID
to confirm Clostridium difficile infection (8) C. difficile 99 (72/73*) 92 (72/78)
• Additional biochemical or molecular tests can be 24h
performed to confirm identification if the specimen has Other culture
not been treated with ethanol. medium 55 (40/73*) 51 (40/78)
48h
QUALITY CONTROL * Number of specimens found to be positive / Number of
specimens found to be positive at 24 hours on chromID C.
Protocol:
difficile and/or at 48 hours on the other medium.
The medium can be tested using the following strains :
®
• Clostridium difficile ATCC BAA2155 The specificities obtained for chromID C. difficile agar
(equivalent to the strain CCUG 60276) (24h) and the other medium (48h) are 88% and 90%
Range of expected results: respectively.

Strain Results at 37°C Specimens inoculated after treatment with ethanol

Out of the 263 specimens tested, 41 were confirmed to be
Clostridium difficile
® positive for C. difficile by PCR.
ATCC BAA2155 Growth and grey-black color
incubation in anaerobic after 24 hours
Recovery %
atmosphere Sensitivity (%)
(*)

Note: chromID™
C. difficile 89 (33/37*) 80 (33/41)
It is the responsibility of the user to perform Quality 24h
Control taking into consideration the intended use of the Other culture
medium, and in accordance with any local applicable medium 81 (30/37*) 73 (30/41)
regulations (frequency, number of strains, incubation 48h
temperature, etc.).
* Number of specimens found to be positive / Number of
specimens found to be positive at 24 hours on chromID C.
LIMITATIONS OF THE METHOD
difficile and/or at 48 hours on the other medium.
• Some micro-organisms other than C. difficile may
develop weakly on the medium producing colonies with The specificities obtained for chromID C. difficile agar
a typical color: C. tertium, C. clostridioforme, (24h) and the other medium (48h) are 98% et and 93%
Bacteroïdes, Lactobacillus. respectively.
• Colonies produced on chromID™ C. difficile agar must
®
not be used with the VITEK 2 ANC identification card
and the ATB™ antimicrobial susceptibility test strips
dedicated to anaerobes (tendency to globally
overestimate resistance). It is recommended to
subculture on a non-selective medium.
• Growth depends on the requirements of each individual
microorganism. It is therefore possible that certain
Clostridium difficile strains which have specific
requirements may not develop.

bioMérieux SA English - 2
chromID™ C. difficile agar (CDIF) 15010 B - en - 2011/03

WASTE DISPOSAL INDEX OF SYMBOLS

Unused reagents may be considered as non hazardous Symbol Meaning
waste and disposed of accordingly.
Dispose of used reagents as well as any other
Catalogue number
contaminated disposable material following procedures for
infectious or potentially infectious products.
It is the responsibility of each laboratory to handle waste In vitro Diagnostic Medical Device
and effluents produced according to their nature and
degree of hazardousness and to treat and dispose of Manufacturer
them (or have them treated and disposed of) in
accordance with any applicable regulations.
Temperature limitation
LITERATURE REFERENCES
1. CROBACH MJT. et al. - European Society of Clinical Use by
Microbiology and Infectious Diseases (ESCMID): Data review
and recommendations for diagnosing Clostridium difficile-
infection (CDI) - Clin Microbiol Infect., 2009; vol. 15, p.1053- Batch code
1066.
2. DELMEE M. et al. - Laboratory diagnosis of Clostridium Consult Instructions for Use
difficile-associated diarrhea: a plea for culture. - J Med
Microbiology - 2005; vol. 54, p.187-191 Microbiology unit,
Louvain University, Brussels, Belgium. Contains sufficient for <n> tests
3. STUART H. et al. – Clinical Practice Guidelines for
Clostridium difficile Infection in Adults: 2010 Update by the
Society fo Health care Epidemiology of America (SHEA) and Protect from light
the Infection Diseases Society of America (IDSA). – Infect
Control Hosp Epidemiol – 2010, vol. 31, p. 431-455.
4. BARTLETT JG., GERDING D. - Clinical Recognition and
Diagnosis of Clostridium difficile Infection - Clin Infect Dis. -
2008;vol. 46, p.S12-S18.
5. DELMEE M., WAUTERS G. - Rôle de Clostridium difficile
dans les diarrhées survenants après antibiothérapie: étude
de 87 cas. - Acta Clin. Belg., 1981, vol. 36, n°4, p. 178-184.
6. GERDING D.N., OLSON M.M., PETERSON L.R. and al. -
Clostridium difficile - associated diarrhea and colitis in adults
- Arch. Intern. Med., 1986, vol. 146, n° 1, p.95-100.
7. Mac GOWAN K.L., KADRE H.A. – Clostridium difficile
infection in children – Clinical Microbiology newsletter, 1999,
vol. 21, n° 7, p. 49-53.
8. RILEY T.V., BOWMAN A., CARROLL S.M. - Diarrhoea
associated with Clostridium difficile in a hospital population -
Med. J. Aust., 1983, vol. 1, n° 4, p. 166-169.
9. ROUSSEAU C. et al. - Comparaison de trois milieux pour la
culture de Clostridium difficile : intérêt des milieux favorisant
la germination des spores ? – Pathol Biol (Paris), 2009,
doi :10.1016/j.patbio.2009.07.001.
10. PERRY D. J. et al. – Evaluation of a Chromogenic Culture
Medium for isolation of Clostridium difficile within 24 hours. –
Journal of clinical microbiology - 2010, p. 3852-3858.
11. RILEY T. V. et al. – Comparison of alcohol shock enrichment
and selective enrichment for the isolation of Clostridium
difficile. – Epidemiol. Infect. – 1987, vol. 99, p. 355-359.

BIOMERIEUX, the blue logo, chromID, VITEK and ATB are used, pending, and/or registered trademarks belonging to bioMérieux SA or one of its
subsidiaries.
ATCC is a trademark belonging to American Type Culture Collection.
CLSI is a trademark belonging to Clinical and Laboratory Standards Institute Inc.
Any other name or trademark is the property of its respective owner.

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