Latex Agglutination Test C-Reactive Protein Latex Test

Introduction Agglutination is the clumping of antigen molecules by specific antibody to the point that the clumping is visible to the human eye. Agglutination assay usually depend on attaching the reagent (eg. antibody) to a larger inert particle such as Latex. Latex particles are inexpensive and appear as a milky solution before agglutination and appear as curdled milk after agglutination. Latex immunoassay provides inexpensive rapid detection methods. C-Reactive Protein or CRP is an acute phase protein produced by the liver during inflammation. CRP binds to pathogens and facilitates phagocytosis. High level of CRP (> 6.0 mg/L) in the serum suggests an acute inflammation in tissue damages, infections and diseases. Progression of a disease or the effectiveness of treatments can be determined by measuring CRP concentration.

Assay Principle Anti-CRP antibodies that are coated with latex particles react with CRP in serum and thus results in agglutination. Latex particle enables the agglutination to be visible. Control samples have two functions; to ensure the test reagents are valid and also to ensure the assay techniques are correct.

Objective 1. To detect the presence of C-reactive protein (CRP) in the serum.

Materials 1. 2. 3. 4. 5. 6. 7. CRP kit Black slide Pipette & dropper Positive control Negative control Latex reagent Serum sample

Method 1. 50 L of serum sample, positive control and negative control was pipetted on the black slide. 2. 1 drop of CRP latex reagent was added to each sample. 3. The slide was mixed and then rotated for 30-60 seconds. 4. Agglutination is checked for all samples. 5. Experiment is repeated using Rheumatoid factor (Rh)

Results Sample Patient A Patient B Patient C Patient D Patient E Discussion Agglutination can be recognized in the positive control well and no agglutination is observed in the negative control well. This suggests that the test reagent is valid and the assay techniques are correct. The test has been set to detect only high levels of CRP (>6.0 mg/L). Lower levels are considered normal. When the patient serum has high CRP level, the Latex will agglutinate and the test will produce a positive result. For a negative result, no agglutination occurs because the CRP level in the sample is lower than 6.0 mg/L. Therefore, from the result obtained, Patient B and Patient D have a high CRP level while Patient A, Patient C and Patient E has low level of CRP in their serum. However, all the results are suggestive and this requires further investigation. RF CRP Test Negative Positive Negative Positive Negative RF Test Positive Negative Positive Negative Positive