C
REACTIVE
PROTEIN
BY: M S RAHMAN
MODERATOR: DR. VARSHA A SINGH
Introduction
2
CRP
An acute phase protein
In response to different inflammatory stimuli
Infection or tissue damage
Produced, mainly in the liver
Extra hepatic - neurons, atherosclerotic plaques,
monocytes, and lymphocytes.(The mechanisms
regulating synthesis - unknown)
Phylogenetically high- plasma protein
Acute-phase protein
Plasma concentration- increases (positive )
- decreases (negative)
- least 25 percent
- inflammatory
disorders.
Positive APP: - CRP,mannose binding protein,
Complement factors,ferritin,
Ceruloplasmin, serum
amyloid A etc.
Negative APP:- Albumin, transferrin, retinol-binding
protein, antithrombin,
transcortin
History
First acute-phase protein
It was discovered by Tillett and Francis in
1930 in the plasma of patients during
the acute phase of pneumococcal
infection.
CRP, named for its capacity to precipitate
the somatic C-polysaccharide of
Streptococcus pneumoniae.
STRUCTURE OF CRP
An annular (ring-shaped),pentameric
protein.
composed of five identical
nonglycosylated polypeptide subunits.
Half-life - 19 hours , constant , health
and disease.
Molecular weight -25106 Da
STRUCTURE OF CRP
Belongs to the pentraxin family of calcium dependent ligandbinding plasma proteins
Production
This is the early and rapid host response to
tissue injury.
Local expansion of pathogen number
Direct activation of compliment in tissues
Degranulation of mast cells
Release of inflammatory mediators
Systemically active mediators:
(IL-1, IL-6, TNF-)
Initiate production of CRP in liver
Activation of C/EBP gene at
Release of CRP in circulation
transcription level
Binding of CRP with phosphocholine
Receptors
Bacterial
cellwall
Apoptotic
Cell
Inflammated
tissue
Phagocytosis of bacteria with the production
of CRP
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CRP PRODUCTION AND KILLING OF
APOPTOTIC CELLS
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CRP PRODUCTION AND KILLING OF
INFLAMMATED TISSUE
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CLINICAL APPLICATION
Hepatic synthesis
Very
rapid, single stimulus.
Healthy young adultMedian
mg/l.
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Serum concentrations rise -
concentration 3.0
6 hr (above 5 mg/l )
-peaks around 48 hours.
RANGE OF CRP LEVELS
VIRAL INFECTIONS: <40mg/l
BACTERIAL INFECTIONS: 40-200 mg/l
SEVERE BACTERIAL INFECTIONS/ TRAUMA/
BURNS: >200 mg/l
Following an acute-phase stimulus-Increase up to 10000-fold
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Acute Inflammation
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A. Acute
inflammation:
Bacterial infection
Pneumococcal pneumonia
Acute rheumatic fever
Bacterial endocarditis
Staphylococcal osteomyelitis
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B. Chronic inflammation:
Systemic lupus erythematosis
Rheumatic arthritis
Reiters syndrome, psoriatic
arthriopathy, arthritis following jejunoileal bypass
Polyarteritis nodosa, disseminated
systemic vasculitis, coetaneous
vasculitis
Polymyalgia rheumatica
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Crohns disease
Ulcerative colitis
Dermomyositis
Osteoarthritis
Neoplastic diseases
Smokers
Obesity
Diabetes
C. Tissue injury:
Tissue injury and surgery
Acute myocardial ischemia
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CLINICAL USES OF CRP
Diagnostic
Prognostic
1. Screening
Infection
Inflammation/ Tissue
damage
Obesity/Hypertensionrisk of CHD
DM type II
Atherosclerosis.
2.Diagnosis of
MeningitisBacterial/ viral
Monitoring of the
response to treatment
of inflammation and
infection.
e.g. : acute pancreatitis
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Lab
Diagnosis
Quantitative
Semiquantitat
ive rapid latex
Agglutination
ELISA
Chemiluminescent
Immunoassay
Laser nephlometry
BNA nephelometer
Quantum dots and
immunochromatogr
aphic test
Immunoturbidimetr
y
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1
Qualitative
Latex
Agglutination
Latex agglutination test
Principle
CRP antigen + mono specific anti-human CRP
Agglutination
Detect greater then 6/ml CRP.
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Semiquantitave
Rapid latex slide test
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Quantitative Method
ELISA
Sandwich Assay
(Peroxidase)
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Quantum dots and immunochromatographic test
Quantum dots (QDs) are introduced as fluorescent probes
and immunochromatographic strips to develop
quantitative fluorescence point-of-care tests (QF-POCT) to
analyze C-reactive protein (CRP) levels
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Rabbit IgG
Human CRP
QD/anti-CRP
conjugate
sample pad
QD-labeled
goatanti-rabbit
Ab
Test line Control line Absorbent pad
CRP Ab2
Fluorescence intensity
Quantum dots and
immunochromatographic test
Distance from sample pad
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Quantitative Method
Chemiluminescent Immunoassay Kits
Use Streptavidin-HRP as
enzyme and enhanced
ECL system as substrate
reagent.
Relative luminosity values
(RLU) is scanned by
photon counter reader
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Quantitative Method
Laser Nephlometry
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Principle
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Light Source: polychromatic tungsten filament
lamp
Monochromator : captures light of multiple
wavelength and changes
to singlewavelength.
(
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Sample: Ag-Ab Complex
interaction of the incident light and
the sample volume
scattered light produced
Photodetector
an electronic signal
converted to a turbidity value.
Immunoturbidimetry-
Ab complex
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Light Source: polychromatic tungsten filament
lamp
Monochromator : captures light of multiple
wavelength and changes
to singlewavelength.
(
3
4
Sample: Ag-Ab Complex
interaction of the incident light and
the sample volume
Absorbed light produced
Photodetector
an electronic signal
converted to a turbidity value.
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5
THANK YOU
