Manual
Description
C-Reactive Protein (CRP) is an acute-phase protein, produced exclusively in the liver. Interleukin-6 is the mediator for the synthesis by the hepatocytes of CRP, a pentamer of approximately 120.000 Daltons. CRP is present in the serum of normal persons at concentrations ranging up to 5mg/l. The protein is produced by the fetus and the neonate and it does not pas the placental barrier, as such it can be used for the early detection of neonatal sepsis.
Because febrile phenoena, leukocyte count and erythrocyte sedimentation rate (ESR) are often misleading, investigators now prefer a quantitative CRP determination as a marker for acute inflammation and tissue necrosis. Within 6 hours of an acute inflammatory challenge the CRP level starts to rise.
Serum concentration of CRP increases significantly in cases of both infectious and non-infectious inflammation, of tissue damage and necrosis and in the presence of malignant tumours. CRP is present in the active stages of inflammatory disorders like rheumatoid arthritis, ankylosing spondylitis, Reiter's syndrome, psoriatric arthropathy, systemic lupus erythematosus, polyarteritis, ulcerative colitis and Crohn's disease.
Widespread malignant disease with carcinoma of the lung, stomach, colon, breast, prostate and pancreas, Hodgkin's disease, non-Hodgkin's lymphoma and lymphosarcoma will give rise to high levels of CRP resulting from tissue damage by invading tumor cells. CRP, therefore may be used to monitor malignancy.
The CRP-level increases dramatically following microbial infections, and this may be particularly helpful for the diagnosis and monitoring of bacterial septicemia in neonates and other immunocompromised patients at risk. In children, CRP is useful for differential diagnosis of bacterial and viral meningitis.
Because the biological half-life of this protein is only 24 hours, CRP accurately parallels the activity of the inflammation process and the CRP concentration decreases much faster than ESR or any other acute phase parameter, which is particularly useful in monitoring appropriate treatment of bacterial diseases with antibiotics.
C-Reactive Protein measurements during the early and late post transplant period of bone marrow and organ transplantations is particularly useful in the management of interfering infections in these immunosuppressed patients.
Typical standard curve
| Standards (µg/ml) |
O.D.value |
| 10 |
2.018 |
| 5 |
1.301 |
| 2.5 |
0.821 |
| 0.5 |
0.240 |
| 0 |
0.019 |
Principle
The human CRP ELISA is a ready-to-use solid-phase enzyme-linked immunosorbent assay based on the sandwich principle with a working time of 1 hour and 10 minutes.
The efficient format of 1 plate with twelve disposable 8-well strips allows free choice of batch size for the assay.
Samples and standards are incubated in microtiter wells coated with antibodies recognizing human CRP.
Peroxidase conjugate antibody will bind to the captured human CRP.
Peroxidase conjugate antibody will react with the substrate, tetramethylbenzidine (TMB).
The enzyme reaction is stopped by the addition of Sulfuric acid.
The absorbance at 450 nm is measured with a spectrophotometer. A standard curve is obtained by plotting the absorbance (linear) versus the corresponding concentrations of the human CRP standards (log).
The human CRP concentration of samples, which are run concurrently with the standards, can be determined from the standard curve.
Use
The human CRP ELISA kit is to be used for the in vitro quantitative determination of human CRP in serum and plasma. This kit is intended for laboratory research use only and is not for use in diagnostic or therapeutic procedures.
Storage and stability
Upon receipt, store individual components at 2 - 8°C. Do not freeze.
Do not use components beyond the expiration date printed on the kit label.
Never use any kit components beyond the expiration date.
Upon receipt, foil pouch around the plate should be vacuum-sealed and unpunctured. Any irregularities to aforementioned conditions may influence plate performance in the assay.
Return unused strips immediately to the foil pouch containing the desiccant pack and reseal along the entire edge of the zip-seal. Quality guaranteed until expiration date if stored at 2 - 8°C.