|
ID-
M. CAT. Cat # 60037
INTENDED
USE
The
Gibson ID- M. CAT. is designed for the detection
of butyrate esterase activity from clinical isolates grown
on culture media as an aid in the presumptive identification
of Moraxella (Branhamella) catanhalls.
SUMMARY
AND EXPLANATION
Indoxyl
butyrate is a substrate which is hydrolyzed by the butyrate
esterase produced by Moraxella catarrhalis.1
Moraxella
catarralis cause infections such as otitis media, sinusitis,
bronchopneumoniae, systemic diseases including endocarditis
and meningitis.2
Different
tests have been reported for the rapid identification of
Moraxella catanhalis, including Tn butynn hydrolysis and
indoxyl butyrate hydrolysis.
PRINCIPLES
The
Gibson ID- M. CAT. utilizes filter paper stnps impregnated
with a substrate for butyrate esterase. This test is based
on the ability of Moraxella catanhalis to hydrolyze the
substance indoxyl butyrate. The enzyme butyrate esterase
breaks the ester linkage between butyryl groups liberating indoxyl. The indoxyl molecules react to form indigo in the
presence of oxygen yielding a distinct bluejireen color
on test cards.
MATERIALS
PROVIDED
20 Test
Cards
Deionized
Water
PRECAUTIONS:
This test is intended for use by those trained in
approprlate laboratory and bacteriological procedures.
This
test is for IN VITRO DIAGNOSTIC USE only. Precautions should
be taken against the dangers of microbiological
hazards. Specimens,
containers, media, and test cards should be sterilized after
use.
STORAGE:
The ID- M. Cat kit must be stored at 2-8C to prevent breakdown
of the substrates and reagents. The kit must be at room
temperature before use, and may be left at room temperature
for up to one hour.
EVIDENCE
OF DETERIORATION:
Test should not be used if the expiration date has
passed. Discard the product if the white filter
paper within the test circle shows signs of
discoloration. If any deficiencies are noted with
this product notify the manufacturer.
SPECIMEN
COLLECTION
Clinical
specimens should be protected from excessive heat and cold
and should be delivered to the laboratory without delay.
The specimen should be collected prior to the initiation
of therapy. If the specimen is collected after the initiation
of therapy, the microbiologist should be notified on the
data form. Additional information on the collection of clinical
specimens may be found in standard reference texts. Fresh
cultures 24-72 hours old give best results. Use pure cultures
of oxidase positive, gram negative dipplococci exhibiting
typical colonial morphology.
PROCEDURE
MATERIALS
REQUIRED BUT NOT PROVIDED:
The standard clinical microbiological
equipment such as loop, burner,
and
incubator are needed for procedures involving the use of
this product. Other materials required include the following:
Oxidase
reagents, Gram Stain Reagents, Microscope slides, Culture
Media, and American Type Culture Collection
(ATCC)
strains- Moraxella catanhalis ATCC 25240 and Neisseria gononhoeae
ATCC 43069.
Use
one card for each specimen tested. Before performing the
test make sure to use a pure culture of oxidase positive,
gram negative, diplococci exhibiting typical colonial morphology.
1.
Remove one test card and add ONE DROP of deionized water
to the test circle. ALLOW 90 SECONDS FOR TEST CARD TO
DRY BEFORE INOCULATION.
2.
Using an applicator stick or Icop, pick up 34 colonies
equal to or greater than 4 mm from a
24-72
hours pure culture and mb on the moistened circle.
3.
Incubate at rcom temperature for up to 5 minutes.
4.
Read color change within the 5 minutes. The formation
of a blue~reen color on the test card is a positive reaction.
The organisms can be presumptively identified as
Moraxella
(Branhamella) catarrhalis.
No color change is a negative test.
INTERPRETATION
USER
QUALITY CONTROL
Daily
quality control should be performed in accordance with
proper laboratory procedures using organisms that will
produce known positive and negative reactions. Follow
steps 14 under PROCEDURE section. If quality control
results are false clinical results should not be recorded.
Notify the manufacturer if proper results are not obtained.
The following American Type Culture Collection strains
are recommended:
Organism
Moraxella
catanhalis ATCC
25240
Neisseda
gononhoeae ATCC
43069
Test
Circle Results
Bluegreen
color change (Positive)
No
color change (Negative)
LIMITATIONS
OF PROCEDURE
Longer
incubation times and excessive moisture may give
false positives, allow test card to dry for 90 seconds
and read test within 5 minutes. Some species of
Staphylococcus (gram positive), Pseudomonas (rods),
and Candida (gram positive) may give a positive
test. Use of only gram negative, oxidase positive,
diplococci should prevent the misidentification
of the above organisms. Animal species, Branhamella
caviae and Branhamella avis, give a positive
result but are rarely encountered in pathological
specimens. False negatives may result from using
tco small an inoculum. The Gibson ID- M.Cat. is
designed as an aid in the presumptive identification
of Moraxella catanhalis. Definitive identification
should be done using established test procedures.2
PERFORMANCE
STUDIES
The
Gibson ID- M CAT. test was tested on 55 dinical
isolates. The isolates consisted of 20 Maraxella
species, 15 Neisseria species, 7 Candida species,
5 Staphylococcus species, and 5 Pseudomonas species.
The Gibson ID- M. CAT. correctly identified all
Moraxella species showing positive results. All
Neisserla strains produced negative results. All
Staphylococcus, Pseudomonas, and two Candida strains
gave positive results. However, it should be noted
that examination of colony morphology, gram-stains,
and oxidase test results should prevent the misidentification
of these isolates.
REFERENCES
1.
Dealer, S.F., et al. 1989. Identification of Branhamella
catarthalis in 2.5 mm with an Indoxyl Butyrate
Strlp Test. J. Clin. Microbial. 27:1390-1391.
2.
Knapp, J.S. and R.J. Rice. 1995. Neisserla and
Branhamella. Pages 324-340 in P.R. Murray, E.J.
Baron, M.A. Pfaller, F.C. Tenaver, and R.H. Yalken,
eds. Manual of Clinical Microbiology. 6th ed.
Amer. Soc. for Microbial., Washington, D.C.
3.
Janda, W.M. and P. Ruther. 1989. B.CAT-CONFIRM,
A Rapid Test for Confirmation of Branhamella catanhalis.
J. Clin. Microbial. 27:1130-1131.
4.
Riou, Y.J. et al. 1981. Hydrolyse De La Trlbutyrlne
Par Les Neisserla et Les Branhamella. Ann. Microbial.
(Inst. Pasteur). 13~:159-169.
5.
Vaneechoutte, M., et al. 1988. Rapid Identification
of Branhamella catarrhalis with 4-Methylumbelliferyl
Butyrate. J. Clin. Microbial. 26:1227:1228.
|
