EXPERIMENT 8 Flashcards
(45 cards)
is a protein found in blood plasma that shows increased levels in response to inflammation
C- reactive protein (CRP)
CRP is synthesized in the liver and has a half – life of (?).
20 to 30 hours
CRP belongs to a group of proteins known as acute phase reactants whose concentrations increase during the course of a general, nonspecific response to (?).
infectious and inflammatory processes, cellular necrosis, and malignant neoplasia
CRP contributes to non – specific immunity by (?).
activating complement and stimulating phagocytosis
Comparing to other acute phase reactants, CRP has the (?).
fastest response time
PRINCIPLE:
Mainly, agglutination.
The C – reactive protein test reagent contains white latex particles coated with (?) and agglutinates in the presence of C – reactive protein in the patient’s serum.
specific antihuman C – reactive protein antibodies
This is a (?) for the qualitative and semi – quantitative detection of C – reactive protein.
rapid slide agglutination test
Example, patient A has CRP present (serve as the antigen) in the serum, if we are going to test the presence of CRP in a patient’s serum, use the reagent (?) with antibodies.
white latex particles that is coated with specific antihuman C- Reactive protein
Since they are specific with each other, (?) will happen.
agglutination
Another example, in Patient B’s serum there is (?).
no CRP
Test the presence of CRP in a patient’s serum, use the reagent (?) with antibodies.
white latex particles that is coated with specific antihuman C-Reactive protein
Since there is no antigen, only the antibody, (?) will happen.
no reaction/ agglutination
Qualitative Test: report if
negative or positive (w/ agglutination)
- Qualitative Test
CRP is found in (?) that is increased in significant levels once there is inflammation or infection
Prepare Reagent (?) and slide
Label the slide as (?)
Bring the reagents and sample to (?) because they are stored at refrigerator temperature
Follow the procedure below
plasma
white latex
positive control, negative control and patient’s sample (serum)
Follow the procedure below
a. Bring the reagents and samples to (?)
b. Place (?) of the control (positive and negative) and (?) of the sample into separate circles on the test card or agglutination slide
c. Resuspend the (?) gently
d. Add (?) to each circle containing the controls and sample/s
e. Mix with disposable (?) and spread over the entire area enclosed by the ring on the test card or agglutination slide. Use a new stirrer for each sample.
f. Rotate the test cards or agglutination slides at (?) (using a mechanical rotator with a tape to hold the slide)
g. Read and interpret results
room temperature
one (1) drop; 50 μL
latex reagent
one (1) drop latex reagent
pipette or stirrer
100 rpm for 2 minutes
Note: if it’s negative, results may be released. But if it’s positive, proceed to the (?) of CRP. Report the titer in semi-quantitative test (actual value of CRP)
semi-quantitative test
Positive sera may be titrated. To titrate, make two – fold serial dilution in normal (?) using the Quantitative Test procedure
saline solution (9g/L saline)
Results:
Positive control has
Negative control and patient’s sample has
clumps
no clumps (homogenous mixture)= no agglutination/ negative
Quantitative Test:
a. Using a semi – automatic pipette, add (?) of normal saline solution to circles (?). Do not spread the saline.
b. Add (?) of patient sample to circle (?)
c. Add (?) of patient sample to circle (?). Mix the sample and saline in circle 2 by drawing the mixture up and down being careful to avoid the formation of any bubbles.
d. Transfer (?) of mixture from circle (?) to the saline in circle (?).
50 μL; 2, 3, 4 and 5
50 μL; 1
50 μL; 2
50 μL; 2; 3
Quantitative Test:
e. Perform serial dilution in the same manner until the last circle, discarding (?) at the end
f. Using the disposable pipette or stirrer, spread the diluted sample over the entire area of each circle starting at circle (?) and moving backwards to the next, reaching until circle (?)
g. Resuspend the latex reagent gently
h. Add (?) latex reagent to each circle containing the sample
i. Mix with disposable pipette or stirrer and spread over the entire area enclosed by the ring on the test card or agglutination slide. Use a new stirrer for each sample.
j. Rotate the test cards or agglutination slides at (?)
k. Read and interpret results.
50 μL
5; 1
one (1) drop
100 rpm for 2 minutes
Guide in the Interpretation of Results:
Examine macroscopically for the presence or absence of clumps or agglutination within (?) after removing the test card or agglutination slide from the rotator
Normal Levels of CRP in Adults: (?)
Negative result: (?)
Positive result: (?)
1 minute
> 6 mg/L
No agglutination would indicate a level of < 6 mg/L
The presence of agglutination indicates a level of ≥ 6 mg/L
CRP concentration (mg/dL) =
limit of sensitivity x titer Limit of sensitivity = 6 mg/L
The (?) is defined as the highest dilution showing macroscopic agglutination
serum titer
