Experiment Number 8 C – REACTIVE PROTEIN (CRP)

Question 1

is a protein found in blood plasma that shows increased levels in response to inflammation.

Answer 1

C- reactive protein (CRP)

Question 2

It is synthesized in the [?] and has a half – life of[?].

Answer 2

liver

20 to 30 hours

Question 3

It belongs to a group of proteins known as [?] whose concentrations increase during the course of a [?] to infectious and inflammatory processes, cellular necrosis, and malignant neoplasia.

Answer 3

acute phase reactants

general, nonspecific response

Question 4

CRP contributes to[?] by activating complement and stimulating phagocytosis

Answer 4

non – specific immunity

Question 5

MATERIALS NEEDED:

Answer 5

Venipuncture set
Test cards or agglutination slides
Micropipette
Pasteur pipette

Question 6

REAGENTS:

Answer 6

CRP latex reagent
Positive control
Negative control
Normal saline solution (0.85 – 0.90% NaCl)

Question 7

SAMPLE:

Answer 7

Serum (should be free from contamination, hemolysis and lipemia)

Question 8

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.

Answer 8

specific antihuman C – reactive protein antibodies

Question 9

This is a rapid slide agglutination test for the [?] of C – reactive protein.

Answer 9

qualitative and semi – quantitative detection

Question 10

Qualitative Test:
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 [?] latex reagent to each circle containing the controls and sample/s
e. Mix with [?] 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 [?]
g. Read and interpret results

Answer 10

room temperature

one (1) drop; 50 μL

latex reagent

one (1) drop

disposable pipette or stirrer

100 rpm for 2 minutes

Question 11

Quantitative Test:
a. Using a semi – automatic pipette, add [?] of normal saline solution to [?]. Do not spread the saline.
b. Add [?] of patient sample to [?]
c. Add also [?] of patient sample to [?]. Mix the [?] in circle 2 by drawing the mixture up and down being careful to avoid the formation of any bubbles.
d. Transfer 50 μL of mixture from circle 2 to the saline in [?].
e. Perform serial dilution in the same manner until the last circle, discarding 50 μL at the end.
f. Using the disposable pipette or stirrer, spread the diluted sample over the entire area of each circle starting at [?] and moving backwards to the next, reaching until [?]
g. Resuspend the latex reagent gently
h. Add one (1) drop 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 100 rpm for 2 minutes
k. Read and interpret results.

Answer 11

50 μL; circles 2, 3, 4 and 5

50 μL; circles 1

50 μL; circles 2

sample and saline

circle 3

circle 5; circle 1

Question 12

Positive sera may be titrated. To titrate, make two – fold serial dilution in [?] using the Quantitative Test procedure

Answer 12

normal saline solution (9g/L saline)

Question 13

Examine macroscopically for the presence or absence of clumps or agglutination within [?] after removing the test card or agglutination slide from the rotator

Answer 13

1 minute

Question 14

Normal Levels of CRP in Adults:

Answer 14

> 6 mg/L

Question 15

✓ Negative result:

Answer 15

No agglutination would indicate a level of < 6 mg/L

Question 16

✓ Positive result:

Answer 16

The presence of agglutination indicates a level of ≥ 6 mg/L

Question 17

CRP concentration (mg/dL) =

Answer 17

limit of sensitivity x titer

Question 18

Limit of sensitivity =

Answer 18

6 mg/L

Question 19

The [?] is defined as the highest dilution showing macroscopic agglutination

Answer 19

serum titer

Question 20

CRP has been detected in serum obtained from apparently

Answer 20

healthy adults and children

Question 21

The reported mean value ranged from

Answer 21

0.1 mg/L in newborns and 0.5 mg/L in male adults

Question 22

The CRP level can increase significantly above the normal levels with the onset of

Answer 22

substantial inflammatory stimulus