CarboFerm™ NEISSERIA KIT

Cat. no. Z98 CarboFerm™ Neisseria Kit 24 tests/kit
Each kit contains:
8-Well Reaction Strips 24 strips
Color Reaction Reader 1 reader
Reaction Worksheets 24 sheets
Inoculation Buffer 24 tubes

INTENDED USE

Hardy Diagnostics CarboFerm™ Neisseria Kit is a rapid test (four hour) for the identification of Neisseria species (including N. gonorrhoeae and N. meningitidis ) and Moraxella ( Branhamella ) catarrhalis . CarboFerm™ uses acid production from carbohydrates and the presence of butyrate esterase to differentiate and identify Neisseria species and M. catarrhalis .

SUMMARY

The majority of Neisseria species are considered normal flora of mucous membranes on humans. Neisseria gonorrhoeae is a sexually transmitted pathogen and Neisseria meningitidis is often associated with meningitis as well as colonization of the nasopharynx. M. catarrhalis causes respiratory tract and other infections.

Neisseria species have traditionally been identified by acid production from carbohydrates. CarboFerm™ Neisseria Kit reduces the time required to determine acid production to four hours (from the usual 48 hours). CarboFerm™ Neisseria Kit rapidly identifies N. gonorrhoeae , N. meningitidis , and M. catarrhalis , as well as other Neisseria species which may be normal flora ( N. lactamica, N. sicca ).

Clinical Laboratory Standards Institute (CLSI - formerly NCCLS) recommends using the butyrate esterase test to identify M. catarrhalis. (9)

FORMULA

Ingredients per liter of deionized water:*

CarboFerm™ Medium Base (well A)
Peptone 2.0gm
Sodium Chloride 1.0gm
Beef Extract 0.6gm
Phenol Red 28.0mg
In addition, the following wells are supplemented with a specific carbohydrate:
Glucose (wells in row C)
Maltose (wells in row D)
Lactose (wells in row E)
Sucrose (wells in row F)



Final pH 6.9 +/- 0.1 at 25ºC.

Well H contains Bromo-Chloro-Indolyl-Butyrate, in an organic solvent, impregnated on a paper disk.


CarboFerm™ Inoculation Buffer
pH Buffers 6.0gm

Final pH 6.9 +/- 0.1 at 25ºC.

* Adjusted and/or supplemented as required to meet performance criteria.

STORAGE AND SHELF LIFE

Storage: Upon receipt store at 2 to 8ºC. away from direct light. The kit should not be used if there is any discoloration or if the expiration date has passed. Do not use if desiccant is missing or if lids have been removed. Product is light and temperature sensitive; protect from light, excessive heat, moisture, and freezing.

PRECAUTIONS

PROCEDURE

Specimen Collection: This product is not intended for primary isolation of patient specimens.

The appropriate organisms for performing the CarboFerm™ Neisseria test are oxidase-positive, gram-negative diplococci.

Method of use:

1. Remove caps from the test strips to be used. Leave strips in the base. Unused strips may be removed from the base and stored in the bag (with caps on) until needed.

2. Use a sterile swab or loop to transfer a pure isolate (from Chocolate, Thayer Martin, or Martin Lewis agar) into a tube of Inoculation Buffer. The culture should be no older than 24 hours. Prepare a heavy suspension (equivalent to a 4.0 McFarland turbidity standard or higher). Vortex and/or aspirate repeatedly until density of the solution is uniform in appearance. A low density may result in false-negative reactions.

3. Aseptically transfer 4 to 5 drops (or approximately 0.18ml) of the suspension into each well of the strip. Do not add any suspension to the second (B) or seventh (G) wells. These wells are intentionally left empty. Do not remove the test strips from the base holder.

4. Incubate strips uncovered at 35ºC. aerobically. Do not incubate in a CO 2 atmosphere. Read the butyrate well (row H) after 10 to 15 minutes. Read the carbohydrate wells (row A, C, D, E, and F) after four hours and before seven hours (some positives will appear in as little as two hours).

INTERPRETATION OF RESULTS

The reaction in row H (butyrate) should be read after 10 to 15 minutes at 35ºC. A blue coloration is considered positive.
Note: Color changes that take place in the butyrate well (H) after 15 minutes of incubation time should be disregarded.

Positive reactions can be read in as little as two hours. Negative carbohydrate wells should be held for a minimum of four hours. A change of color from red to orange or yellow is considered positive for the carbohydrate wells (wells C through F). Well A is used as a reference and should remain negative (red) throughout incubation, since it contains no carbohydrate. Read final results at four hours.
Note: Color changes in the carbohydrate wells that take place after seven hours of incubation time should be disregarded.

A carbohydrate reaction which is more yellow or orange than the control well (A) is considered positive.

Use the chart below to identify the listed organisms.

Organism Negative
Control
(well A)



Glucose
(well C)
Maltose
(well D)
Lactose
(well E)


Sucrose
(well F)


Butyrate
(well H)


N. gonorrhoeae / cinerea* -
red
+
orange or
yellow

-
red
-
red
-
red
-
white
N. meningitidis -
red
+
orange or
yellow

+
orange or
yellow

-
red
-
red
-
white
N. lactamica -
red
+
orange or
yellow

+
orange or
yellow

+
orange or
yellow

-
red
-
white
N. sicca / N. mucosa** -
red
+
orange or
yellow

+
orange or
yellow

-
red
+
orange or
yellow

-
white
M. catarrhalis -
red
-
red
-
red
-
red
-
red
+
blue

* Some strains of N. cinerea can be weakly positive for glucose. This has not been observed with CarboFerm™. Note that N. cinerea will not usually grow on Thayer Martin Agar (since it is usually sensitive to colistin) but will grow on Nutrient Agar. N. gonorrhoeae will grow on Thayer Martin Agar (Cat. no. E30) but will not grow on Nutrient Agar (Cat. no. L20).

** To distinguish between N. sicca and N. mucosa , perform a nitrate reduction test (Cat. no. K42). N. mucosa is positive for nitrate reduction and N. sicca is negative.

LIMITATIONS

This product is intended for pure cultures of oxidase-positive, gram-negative diplococci.

Some strains may give a weak positive reaction which is seen as an orange color in the wells containing carbohydrates or a pale blue in the butyrate well. Carbohydrate reactions should be considered positive if and only if the well containing the carbohydrate is more orange/yellow than the negative control well (row A).

Butyrate reactions should be read within 15 minutes or erroneous results may occur.

False-negatives may result from using too small of an inoculum, or old cultures (greater than 48 hours).

MATERIALS REQUIRED BUT NOT PROVIDED

Standard microbiological supplies and equipment such as loops, culture media, transfer pipettes, swabs, applicator sticks, incinerators, and incubators etc., as well as serological and biochemical reagents, are not provided.

QUALITY CONTROL

Test Organisms Well Incubation Results
Time Temperature Atmosphere
N. gonorrhoeae
ATCC ® 43069
Control
Glucose
Maltose
Lactose
Sucrose
Butyrate




4hr
4hr
4hr
4hr
4hr
10-15min




35°C Aerobic Red
Positive, yellow/orange
Negative, red
Negative, red
Negative, red
Negative, white




N. meningitidis
ATCC ® 13090
Control
Glucose
Maltose
Lactose
Sucrose
Butyrate




4hr
4hr
4hr
4hr
4hr
10-15min




35°C Aerobic Red
Positive, yellow/orange
Positive, yellow/orange
Negative, red
Negative, red
Negative, white




N. lactamica
ATCC ® 23970
Control
Glucose
Maltose
Lactose
Sucrose
Butyrate




4hr
4hr
4hr
4hr
4hr
10-15min




35°C Aerobic Red
Positive, yellow/orange
Positive, yellow/orange
Positive, yellow/orange
Negative, red
Negative, white




N. sicca
ATCC ® 9913
Control
Glucose
Maltose
Lactose
Sucrose
Butyrate




4hr
4hr
4hr
4hr
4hr
10-15min




35°C Aerobic Red
Positive, yellow/orange
Positive, yellow/orange
Negative, red
Positive, yellow/orange
Negative, white




M. catarrhalis
ATCC ® 25240
Control
Glucose
Maltose
Lactose
Sucrose
Butyrate




4hr
4hr
4hr
4hr
4hr
10-15min




35°C Aerobic Red
Negative, red
Negative, red
Negative, red
Negative, red
Positive, blue




Check for signs of deterioration. It is recommended that users perform quality control testing with known organisms to demonstrate a positive reaction and a negative reaction.

Physical Appearance

CarboFerm™ Neisseria Kit

CarboFerm™ Neisseria Kit (Cat. no. Z98).

CarboFerm™ Neisseria Kit

Showing all results for the CarboFerm™ Neisseria Kit (Cat. no. Z98).

N. gonorrhoeae

Showing results for Neisseria gonorrhoeae (ATCC ® 43069) with the CarboFerm™ Neisseria Kit (Cat. no. Z98).

N. meningitidis

Showing results for Neisseria meningitidis (ATCC ® 13090) with the CarboFerm™ Neisseria Kit (Cat. no. Z98).

N. lactamica

Showing results for Neisseria lactamica (ATCC ® 23970) with the CarboFerm™ Neisseria Kit (Cat. no. Z98).

N. sicca

Showing results for Neisseria sicca (ATCC ® 9913) with the CarboFerm™ Neisseria Kit (Cat. no. Z98).

M. catarrhalis

Showing results for Branhamella (Moraxella) catarrhalis (ATCC ® 25240) with the CarboFerm™ Neisseria Kit (Cat. no. Z98). Note that the first well is blue.

REFERENCES

1. Anderson, N.L., et al. Cumitech 3B; Quality Systems in the Clinical Microbiology Laboratory, Coordinating ed., A.S. Weissfeld. American Society for Microbiology, Washington, D.C.

2. Tille, P., et al. Bailey and Scott's Diagnostic Microbiology, C.V. Mosby Company, St. Louis, MO.

3. Berger, U. 1962. Uber die spaltung von tributyrin durch Neisseria. Arch Hyg. Bakteriol.; 146:388-391.

4. Isenberg, H.D. Clinical Microbiology Procedures Handbook, Vol. I, II & III. American Society for Microbiology, Washington, D.C.

5. Janda, W.M. and P. Ruther. 1989. B. CAT CONFIRM; A rapid test for confirmation of Branhamella catarrhalis. J. Clin. Microbiol.; 27:1390-1391.

6. Koneman, E.W., et al. Color Atlas and Textbook of Diagnostic Microbiology, J.B. Lippincott Company, Philadelphia, PA.

7. Jorgensen., et al. Manual of Clinical Microbiology, American Society for Microbiology, Washington, D.C.

8. Perez, J.L., et al. 1990. Butyrate esterase (tributyrin) spot test, a simple method for immediate identification of Moraxella (Branhamella) catarrhalis. J. Clin. Microbiol.; 28: 2347-2348.

9. Abbreviated Identification of Bacteria and Yeast; Approved Guideline, M35A, Vol. 22, No. 18. 2002. Clinical Laboratory Standards Institute (CLSI - formerly NCCLS), Villanova, PA.


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