Part of Thermo Fisher Scientific
Introduction
Clostridium difficile is recognised as a major cause of infectious
diarrhoea in hospital patients. Infection with the organism is commonly responsible
for pseudomembranous colitis and antibiotic associated diarrhoea, as well
as post-operative diarrhoea1. The isolation and identification of Clostridium
difficile is important in establishing the aetiology of these pathologies.
Methods for the selective isolation of Clostridium difficile have been described 2
and the rate of isolation is increased by culturing on selective media7. However,
subsequent identification of the organism has required the use of gas liquid
chromatography since conventional biochemical tests are often of little value.
Although detection of Clostridium difficile cytotoxin is widely used in diagnosis, the
association between the presence of cytotoxin and the symptoms of infection
is inconsistent 3,4.
The Oxoid C. difficile Test Kit is a rapid and
simple latex agglutination test for the early identification of Clostridium difficile and
is ideally suited to screening selective and enrichment broth cultures. The test
can also be used
for the identification of Clostridium difficile colonies from selective solid
media 2,6. The recommended procedure for the identification of Clostridium
difficile using this
kit is compared with a typical procedure below:
A comparison of methods for Isolation and identification of Clostridium difficile
Time |
Oxoid Suggested Procedure |
Conventional Laboratory Procedure |
||
Faeces |
Faeces |
|||
24 hours |
Selective enrichment broth |
Alcohol shock Procedure Enrichment broth 37°C, 18-24 hours |
||
Test with Oxoid C.difficile latex (2 mins) |
Report |
|||
Negative Presumptive Report |
Positive Report |
|||
48 hours |
Subculture onto selective Medium (Clostridium Difficle
agar base + selective supplements - SR0096 or SR0173) 37°C Anaerobically, 24-48 hours |
Subculture
onto selective medium (CCFA) |
||
Test with Oxoid C.difficile latex (2 mins) |
Incubate at 37°C Anaerobically 48-72 hours |
|||
Negative |
Confirmed Positive Report |
|||
72 hours |
Examine
morphologically forClostridium difficile |
|||
Negative |
Positive Report |
|||
Gram Stain |
Long Wave UV light |
G.L.C. (Approx 30 minutes) Set up biochemical Identification (24-48 Hours) |
||
Read Results |
The advantages of using the Oxoid C. difficile Test Kit are:
1. Earlier identification of Clostridium difficile in primary selective cultures
allowing presumptive positive identification overnight, and confirmed positive
results
within two days. Identification by anaerobic culture and confirmatory tests
requires 3-6 days.
2. The high predictive value of a negative result allows early elimination
of negative selective cultures.
3. Reduction in the required number of subcultures.
4. No requirement for extended anaerobic subcultures of 48-72 hours.
5. No requirement for expensive equipment.
Principle of the Test
Latex particles are coated with IgG antibodes specific for Clostridium difficile cell
wall antigens. When mixed on a reaction card with selective or enrichment
broth containing the organism, or with a suspension of Clostridium difficile colonies
from solid media, the latex particles agglutinate in large visible clumps
within 2 minutes.
Components of the Kit
C.difficile Reagent:
Latex particles coated with rabbit IgG against Clostridium difficile; preserved
with 0.02% merthiolate (2.5 ml)
Positive Control: Inactivated C. difficile (0.5 ml)
0.85% isotonic saline, preserved with 0. 1 % sodium azide (5 ml)
Disposable reaction cards
Disposable mixing sticks
Materials required but not provided
Pasteur pipettes
For full procedure please see product insert.
Storage
The Oxoid C. difficile Test Kit should be stored at 2- 8°C. Do not freeze.
The kit should not be used after the expiry date printed on the outside of
the
carton.
Precautions
The Positive Control contains inactivated Clostridium difficile but should
be treated as a potentially infectious agent.
The isotonic saline is preserved with sodium azide which can form potentially
explosive compounds on contact with copper and lead plumbing. On disposal of
reagent, flush with copious quantities of water to prevent azide build up.
Reference
1. Larson, H.E., Price, A.B., Honour, P. and Borriello, S.P. (1978) Lancet 8,
1063 1066
2. George, W.L., Sutter, VL, Citron, D. and Finegold, S.M. (1979) J.Clin.
Microbiol. 9, 214 219
3. Riley, TV., Bowman, R.A. and Carroll, S.M. (1983) Med. J. Aust i, 166 169
4. Haslam, S.C., Ketley, J.M., Mitchell, T.J., Stephen, J, Burdon, D.W. and
Candy, D.C.A. (1986) J. Med. Microbiol. 21, 293 297
5. Mollby, R, Nord, C.E. and Aronsson, B. (1980) Scand. J. Infect. Dis. 22
(Suppl), 30 36
6. Carroll, S.M., Bowman, R.A. and Riley, TV. (1982) Pathol. 15, 165 167
7. Bowman, R.A., Arrow, S.A. and Riley, T.V. (1986) J. Clin. Pathol. 39, 212
214
8. Chang, T.W. and Gorbach, S. L. (1982) J. Clin. Microbiol. 15, 465 467