CAMP Spot Test Reagent
Hardy Diagnostics CAMP Spot Test Reagent is used to rapidly determine an organism's ability to produce the protein CAMP factor. CAMP factor is a diffusible, extracellular protein which acts synergistically with staphylococcal beta-hemolysin (beta-lysin). The test is particularly useful as part of the identification of Streptococcus agalactiae (group B streptococci), and Listeria monocytogenes.
The CAMP test (named for the original authors: Christie, Atkins, and Munch-Petersen) was first used in the identification of group B streptococci (GBS). Group B streptococci secrete a protein called CAMP factor or "protein B" that interacts with the beta-hemolysin produced and secreted by Staphylococcus aureus, this results in enhanced or synergistic hemolysis.A variety of methods are currently available to identify GBS isolated from clinical specimens. These methods can be time consuming and/or expensive compared to the CAMP spot test. Hardy Diagnostics CAMP Spot Test Reagent is used as a rapid CAMP test method. The reagent, containing staphylococcal beta-lysin (also called beta-toxin, beta-hemolysin, or beta-staphylolysin), acts directly with the CAMP factor that is diffused into the medium around the suspect colony. The beta-lysin has a synergistic effect in the presence of CAMP factor, producing enhanced hemolysis of sheep erythrocytes. Enhanced hemolysis is visible within 30 minutes to one hour of placing a drop of CAMP Spot Test Reagent next to an isolated beta-hemolytic colony.Group B streptococci (GBS) are associated with a broad spectrum of clinical syndromes, including neonatal sepsis and meningitis. It is the current CDC recommendation that laboratories culture for the organism during the latter part of the prenatal period, as part of an effort to prevent neonatal GBS infections.Of the Listeria species, L. monocytogenes is the only one that has been clearly documented as a human pathogen and is one of the few species of Listeria that is CAMP factor positive. Listeria monocytogenes has been associated with sepsis and meningitis in elderly and other immunocompromised persons. It can cause a flu-like illness in pregnant females, and, if untreated, can infect the fetus with serious sequellae. Contaminated foods, such as dairy products, coleslaw, meat, etc. are the primary vehicle of infections.Production of the CAMP factor is indicative of group B streptococci and Listeria monocytogenes, when combined with the appropriate colony morphology, gram reaction and catalase test result. In addition to performing the CAMP spot tests on hemolytic colonies on sheep blood agar, the test can be performed on non-hemolytic strains of GBS and on GBS growing on Columbia CNA with sheep blood agar. The CAMP spot test can also be performed on the same blood agar plate with the Bacitracin and SXT disk tests. Since the GBS colony is not disturbed during the spot test, the colony can be used for additional testing.
CAMP Spot Test Reagent contains a portion of filtrate from a culture of beta-lysin producing Staphylococcus species.
The CAMP spot test can be performed on a single colony from a primary isolation sheep blood agar plate or Columbia CNA with sheep blood. The test can be done as soon as a colony becomes visible on the plate, and as long as the colony has been growing for at least 18 hours.
1. Remove one vial from the freezer and allow it to thaw.
2. Using a pipette, place one drop of CAMP Spot Test Reagent next to a characteristic colony grown for 18-24 hours on a Blood Agar plate (Cat. no. A10) or Columbia CNA plate (Cat. no. A50). The liquid may touch or even engulf the colony.
Note: Manufacturers of commercially prepared culture media are not required to verify that Columbia CNA with sheep blood will provide a positive CAMP reaction. Users should test each lot of Columbia CNA with a positive and negative control prior to using the media for the CAMP Spot Test.
3. Incubate the plate aerobically or in 5-10% CO2 at 35 degrees C. (right side up so that the reagent won't run over the plate's surface) for 20 to 30 minutes.
4. Observe, using transmitted light, for an arc or circle of enhanced hemolysis next to the colony. If reaction is negative, reincubate for an additional 30 minutes.
Note: Refrigeration after the initial incubation period may enhance the reaction.
INTERPRETATION OF RESULTS
A positive result is indicated by the presence of a clear zone (arc or circle) of enhanced hemolysis. Enhanced hemolysis only where the diffused, slight hemolysis overlaps is considered a positive reaction. A negative result will show no areas of enhanced hemolysis near the colony in the presence of the CAMP Spot Test Reagent.CAMP test reactions for Listeria species:
* Listeria monocytogenes, ATCC® 15313, is atypical with respect to its CAMP reaction compared to other strains and does not give a positive CAMP reaction with S. aureus.
Cat. no. Z206
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