(Z1…See I)- {{{ CAMP test was first identified by Christie, Atkins, and Munch-Peterson in 1944 and CAMP

test is an acronym of three

researchers.

It is used for the presumptive identification of Group B Streptococci (Streptococcus agalactiae). It is the only beta-hemolytic Streptococcus
which secretes a protein called CAMP factor or “protein B”. CAMP test rarely give false positive with other Streptococci. }}}

(Z2) Principle

CAMP test detects the production of diffusible, thermostable, extracellular protein known as CAMP factor, produced by Group B
Streptococcus.

The CAMP factor acts synergistically with the beta lysin produced by Staphylococcus aureus to produce a zone of enhanced lysis of sheep
or bovine erythrocytes.

The standard CAMP test depend on the elaboration of two toxins during growth to form a typical arrowhead or flame-shaped clearing at
the junction of the two organisms when they are placed perpendicular to each other.

(Z3) Purpose of CAMP test

This test is useful in the identification of both S. agalactiae and many gram-positive rods, including Listeria monocytogenes.

(Z4) Procedure of CAMP Test

A. Standard CAMP test

1. Using an inoculating loop, streak a beta-lysin-producing Staphylococcus aureus (ATCC25923) in a straight line across the center of a
sheep blood agar plate.
2. Streak test organism in a straight line perpendicular to the S. aureus leaving 1cm space between the two streaks. (Multiple organisms
can be tested on a single plate if they are 3 to 4mm apart).
3. Incubate the plate at 37 degree Celsius in ambient air for 18-24 hours.

B. CAMP Spot test

1. Remove test kit 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 blood agar plate.
3. Incubate the plate aerobically or in 5-10% CO2 at 37°C for 20-30 minutes.
4. Observe, using transmitted light, for an arc or circle of enhanced hemolysis next to the colony. If reaction is negative, re-incubate for an
additional 30 minutes.

Result Interpretation of CAMP Test

Positive: Enhanced hemolysis is indicated by an arrow head-shaped zone of beta-hemolysis at the junction of the two organisms.
In CAMP Spot test positive result is indicated by the presence of clear zone (arc or circle) of enhanced hemolysis.

Negative: No enhancement of hemolysis.

In CAMP Spot test negative result shows no area of enhanced hemolysis near the colony in the presence of test reagent.

NOTE: A similar test has been described for Listeria ivanovii, where an “arrowhead” hemolysis occurs appear between streaks of Listeria
ivanovii and Rhodococcus equi.

Quality control

Positive: Streptococcus agalactiae ATCC 13813

Negative: Streptococcus pyogenes ATCC19615

CAMP Test for the identification of Listeria monocytogenes

A version of the CAMP test was first used by Groves to identify Listeria monocytogenes. He found that the pathogenic Listeria
monocytogenes was also positive for CAMP test. Listeria monocytogenes is a biosafety level 2 organism and should be handled with
precautions.

To perform CAMP test for the identification of Listeria monocytogenes, Listeria monocytogenes is streaked at right angle to the streak of
beta-hemolytic Staphylococcus aureus on sheep blood agar plate. Then incubate the palte in ambient air at 37 degree Celsius for 24 hours.
Positive CAMP: Enhanced zone of beta-hemolysis and a smaller less obvious rectangular zone of hemolysis.

Limitation of CAMP test

1. Some Group A Streptococcal will be CAMP test positive if the plate is incubated in a candle jar in an atmosphere or under anaerobic
conditions. Therefore, ambient air incubation should be done.
2. Extended incubation times or elevated incubation temperatures may give false-positive results.
3. Sheep blood agar plates are only used. Human, horse, rabbit, or guinea pig blood plates will not give a proper reaction.
4. L. ivanovii only shows a positive CAMP reaction when using an alternative CAMP test method, in which Rhodococcus equi replaces S.
aureus.
5. Colonies of Listeria monocytogenes have a narrow zone of beta-hemolysis on sheep blood agar and may be confused with group B
beta-hemolytic streptococci, if catalase and gram stain are not performed.