C-reactive protein: cards test
c-reactive protein is a serum protein which is synthesized in the liver
it's rate of synthesis and secretion increases within hours of an acute injury or the onset of
inflammation normally it is present only in trace amounts in serum but it can increase as much
as thousandfold in response to injury or infection
the clinical measurement of CRP in serum therefore is a sensitive index of disease activity in
inflammatory
infective
ischemic conditions
the CRP reagent kit is based on the principle of latex agglutination assay
Principle
the CRP reagent kit is based on an immunological reaction between antibodies against CRP bound to
biologically inert latex particles and CRP in the test specimen. When Sivan containing CRP greater
than 0.6 milligrams per deciliter(mg/dL) is mixed with the latex reagent visible agglutination occurs.
Reagents
1. CRP reagent, it is a uniform suspension of polystyrene latex particles coat it with anti CRP
antibodies
2. positive control
3. negative control
reagent storage and stability
store the reagent at 2 to 8 degree centigrate
do not freeze
do not use reagents after the expiry date
additional material required
stopwatch
test tubes
isotonic saline
Pippets and tips
mixing sticks
specimen collection and preparation
no special preparation of the patient is required prior to specimen collection by approved
techniques
only serum must be used for testing if a delay in testing occurs
store the sample at 2 to 8 degree centigrate
Procedure
Bring reagent and samples to room temperature before testing
Qualitative method
Place one drop of undiluted test sample on field two
place one drop or approximately forty microliters of CRP positive control on field one of the
reaction slide
place one drop of the CRP negative control on to field three
add one drop of CRP latex reagent to all the fields
do not let the dropper tip touch the liquid on the slide
using a mixing stick mix the test specimen and latex reagent uniformly over the entire circle
immediately start a stopwatch
rub the slide gently back and forth observing for agglutination macroscopically at two minutes
Interpretation off results
Agglutination is a positive test result and indicates presence of detectable levels of CRP in the
test specimen
No agglutination is a negative test result and indicates absence of detectable levels of CRP in
the test specimen
Semi-quantitative method
1. Using isotonic saline prepare serial dilutions of the test specimen positive in the qualitative
method
2. take a reaction slide and label it with patient identification
3. mark reaction circle 1 as positive control (PC) and the remaining circles from 2 to 6 as 1:2 , 1:4 ,
1:8 1:16 , 1:32 for the respective dilutions
4. place 50 microliters of 0.9% saline in circles numbered two through six
5. place one drop of positive control on reaction circle 1
6. place 50 microliters of serum in circle two
7. mix thus a line and the serum in circle 1 by drawing the mixture up and down this gives 1 into
dilution
8. transfer 50 microliters from circle 2 to circle 3 and mix
9. transfer 50 microliters from circle 3 to circle 4 and mix
10. transfer 50 microliters from circle 4 to circle 5 mix
11. transfer 50 microliters from circle 5 to circle 6 and mix
12. discard the last 50 microliters
13. add one drop of CRP latex reagent to all the reaction circles
14. using a mixing stick mix the test specimen and the latex reagent uniformly over the entire Circle
immediately start a stopwatch
15. Rock the slide gently back and forth observing for agglutination macroscopically at 2 minutes
Interpretation of results
agglutination in the highest serum dilution corresponds to the approximate amount of CRP in
milligrams per deciliter present in the test specimen
concentration of CRP can be calculated as follows
CRP (mg/dl) = S x D
where
S = sensitivity of the reagent ………. 0.6 mg/dl
D= highest dilution of the serum showing agglutination
Precautions
all standard precautions should be adhered to while handling the test samples and performing the
tests
do not use markedly
Lipemic
hemolyzed
contaminated serum samples
use of plasma rather than serum can lead to false positive results
do not reach results beyond indicated testing time limits
the reagents contain 0.1% sodium azide as preservative
avoid contact with skin and mucosa on disposal flush with large quantities of water
store the reagents according to manufacturer's instructions
performance of the reagent must be verified with the positive and negative controls provided
with the kit
shake the latex reagent well before use
do not use damaged or leaking reagents
