Group 1 BSMT 3A Act. 5.2

LEADER David, Alyssa Deanne DATE April 24, 2021
MEMBERS Almoite, Kimberly

Landingin, Kyla Andrea
Munoz, Antonnette
Pineda, Tracy
Sicat, Trisha Lynne
SECTION BSMT- 3A GROUP 1
Activity 5.2
C-Reactive Protein
OBJECTIVE: To be able to perform the qualitative and semi quantitative measurement
of the antibodies to CRP - a serum protein which is synthesized in the liver. Its rate of
synthesis and secretion increases within hours of an acute injury or the onset of
inflammation.
EQUIPMENTS/ REAGENTS NEEDED:

● Rhelax CRP reagent a suspension of polystyrene particles coated with
● anti CRP antibodies (monoclonal IgG)
● NSS
● CRP positive control - a stabilized human serum reactive in test system
● CRP negative control - a stabilized human serum non-reactive in test system
● Glass slide
● Stirring rod
● 1 serological pipette (1ml)
● 7 test tube for dilutions
● 1 test tube rack
PRINCIPLE:
The Rhelax CRP slide test for detection of CRP is based on the principle of agglutination.
The serum is mixed with CRR reagent and allowed to react. If CRP concentration is
greater than 0.6 mg/dl, a visible agglutination is observed.
PROCEDURE:
QUALITATIVE METHOD:
1. Place one drop of serum on the glass slide using a disposable pipette provided in
the kit
2. Add one drop of Relax CRP latex reagent to the drop of specimen on the slide. Do
not let the dropper tip touch the liquid on the slide.
3. Using a mixing stick, mix the serum and the reagent uniformly over the entire
circle
4. Immediately start a stopwatch. Rock the slide gently back and forth, observing for
agglutination macroscopically at two minutes.
Interpretation of Results:
(+) = presence of agglutination and indicates the presence of CRP in the serum
(-) = no agglutination
SEMI - QUANTITATIVE METHOD:

1. Using isotonic saline (NSS) prepare serial dilutions of the serum sample.
TUBE NUMBER VOLUME OF VOLUME OF NSS DILUTION

SERUM
1 0.1 ml 0.1 ml 1:2
2 0.1 mL from slide 1 0.1 ml 1:4
3 0.1 mL from slide 2 0.1 ml 1:8
4 0.1 mL from slide 3 0.1 ml 1:16
5 0.1 mL from slide 4 0.1 ml 1:32
6 0.1 mL from slide 5 0.1 ml 1:64
2. Perform qualitative test for each tube on the slide

3. Observe for agglutination in the highest serum dilution corresponds to
approximate amount of CRP in mg/dL [D]
4. Concentration of CRP can be calculated as follows: CRP = 0.6 x D
DRAW AND ILLUSTRATIONS OF PROCEDURE
QUESTIONS FOR RESEARCH:
1. Why is plasma not satisfactory for the determination of CRP?
CRP is phylogenetically a highly conserved plasma protein, with homolog
in vertebrates and many invertebrates that participates in the systemic
response to inflammation. Its plasma concentration increases during
inflammatory states, a character that has long been employed for clinical
purposes. CRP is a pattern recognition molecule, binding to specific molecular
configurations that are typically exposed during cell death or found on the
surfaces of pathogens. Its rapid increase in synthesis within hours after tissue
injury or infection suggests that it contributes to host defense and that it is part
of the innate immune response.
2. Explain why CRP is never diagnostic? Give medical significance of CRP

CRP can never be diagnostic on its own and can only be interpreted at the
bedside, in full knowledge of all other clinical and pathologic results.
However, it can then contribute powerfully to management, just as recording
of the patient’s temperature, an equally nonspecific parameter, is of great
clinical utility. The CRP test is not diagnostic, but it provides information to
your healthcare practitioner as to whether inflammation is present, without
identifying the source of the inflammation.
- Medical significance:
A c-reactive protein test measures the level of c-reactive protein (CRP) in
your blood. CRP is a protein made by your liver. It's sent into your
bloodstream in response to inflammation. Inflammation is your body's way of
protecting your tissues if you've been injured or have an infection. In
Transplant cases, Elevated levels were seen in majority of kidney or heart
transplant – Highly elevated in GVHD – Changes in levels are not organ
specific, like other inflammatory conditions
3. What is the advantage of CRP over ESR determination?
Here are the following advantages of C-reactive protein (CRP) over ESR
determination:
❖ C-reactive protein has been found to be a better marker of the acute phase
reaction than the erythrocyte sedimentation rate
❖ C-reactive protein was considered more sensitive test and rapidly detects
changes in the acute phase reaction.
❖ In terms of Differentiation of borderline cases from positive and negative
cases CRP has true differentiation while ESR has no true differentiation
and has an Unspecific screening.
❖ CRP is suitable for therapeutic monitoring with given, fast reaction, while
ESR is limited and slow.
❖ Contamination risk of CRP is minimized while ESR it is dependent on test
tube.
4. Describe physical and chemical nature of CRP
CRP plays an essential role in the recognition of self and foreign

molecules. This interaction leads to an activation of the adaptive immune
system early in the course of an inflammatory or infectious process. CRP is
considered the prototypical acute phase reactant in man. Acute phase reactants
are proteins that are produced by the liver in response to a variety of clinical
conditions including infection, inflammation and trauma. Clinically, CRP is
used as a monitor of the inflammatory response in diseases such as
rheumatoid arthritis and vasculitides. The production of CRP has also been
used to monitor for the presence of infection and its successful treatment. The
major stimulus to its secretion by the liver is the pro inflammatory cytokine,
interleukin (1L)-6. CRP is a member of the pentraxin family of proteins.
CRP was first identified by its reactivity with the C polysaccharide of the
pneumococcus and was thus named the C-reactive protein. The binding of
CRP to the C-polysaccharide was found to be calcium dependent. This
characteristic has been used to determine the specificity of its binding to other
ligands. One of the first recognized natural ligands of CRP was the
phosphatidylcholine present on damaged cell membranes. Later it was
determined that CRP could bind to chromatin when it was exposed or
denatured, as might occur during apoptosis or necrosis. CRP also binds to the
small nuclear ribonucleoprotein (snRNP) particles. Although the nature of the
ligand bound is not known, CRP is localized to inflammatory sites in vivo
where there is acute inflammation, often with polymorphonuclear leucocytes
(PMN) colocalized.
5. Name other methods of CRP determination
❖ Rate Nephelometry
- Nephelometry provides accurate and precise quantitation of serum

proteins, and because of automation the cost per test is typically lower
than other methods. Additionally, very small samples can be analyzed.
❖ Reverse passive agglutination testing

- has also been used to measure levels of certain therapeutic drugs,
hormones, and plasma proteins such as haptoglobin and C-reactive
protein.
References:
Du Clos, T. W. (2000). Function of C-reactive protein. Retrieved April 18, 2021, from
https://www.tandfonline.com/doi/pdf/10.3109/07853890009011772
GmbH, D. D. S. (n.d.). Advantages of CRP. CRP IS. Retrieved April 18, 2021,
from https://www.crp- is.com/advantages-of-crp/
Harrison, M. (n.d.). Erythrocyte sedimentation rate and C-reactive protein. NPS

MedicineWise.
Retrieved April 18, 2021, from https://www.nps.org.au/australian-
prescriber/articles/erythrocyte-sedimentation-rate-and-c-reactive
Faraj, M. and Salem, N. (September 21st 2012). C-Reactive Protein, Blood Cell - An
Overview of Studies in Hematology, Terry E. Moschandreou, IntechOpen, DOI:
10.5772/47735. Available from: https://www.intechopen.com/books/blood-cell-
an-overview-of-studies-in-hematology/c-reactive-
protein?fbclid=IwAR2_4RHLtFyDgAwl24pM3UW8fWpU5d-
ubCH1iQrqRnALCnIewGXGrt2bZiU
Turgeon, M.L. (2013). Agglutination Methods. Immunology & Serology in Laboratory

(5th ed., Vol. 5). Elsevier Inc.
Miller, L. E. (2017). Precipitation and Agglutination Reactions. Clinical Immunology and

Serology, Fourth Edition (4th ed.) Pg. 143-149. F.A Davis Company.
LabTestOnline (2019) C-Reactive Protein (CRP) retrieved from:

https://labtestsonline.org/tests/c-reactive-protein-crp
Hirschfield G., Pepys M. (2015) C-reactive protein: a critical update retrieved from
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC161431/

Group 1 BSMT 3A Act. 5.2

Uploaded by

Copyright:

Available Formats

Group 1 BSMT 3A Act. 5.2

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Group 1 BSMT 3A Act. 5.2

Uploaded by

Copyright:

Available Formats

LEADER David, Alyssa Deanne DATE April 24, 2021

MEMBERS Almoite, Kimberly

EQUIPMENTS/ REAGENTS NEEDED:

SEMI - QUANTITATIVE METHOD:

TUBE NUMBER VOLUME OF VOLUME OF NSS DILUTION

2. Perform qualitative test for each tube on the slide

2. Explain why CRP is never diagnostic? Give medical significance of CRP

3. What is the advantage of CRP over ESR determination?

4. Describe physical and chemical nature of CRP

CRP plays an essential role in the recognition of self and foreign

5. Name other methods of CRP determination

- Nephelometry provides accurate and precise quantitation of serum

❖ Reverse passive agglutination testing

Harrison, M. (n.d.). Erythrocyte sedimentation rate and C-reactive protein. NPS

Turgeon, M.L. (2013). Agglutination Methods. Immunology & Serology in Laboratory

Miller, L. E. (2017). Precipitation and Agglutination Reactions. Clinical Immunology and

LabTestOnline (2019) C-Reactive Protein (CRP) retrieved from:

You might also like