Create a 5 page essay paper that discusses INFECTION & IMMUNITY PRACTICAL : DETECTION OF ANTIVIRAL ANTIBODIES IN SERUM USING AN ELISA TECHNIQUE.Download file to see previous pages... Antibody is produ

Create a 5 page essay paper that discusses INFECTION & IMMUNITY PRACTICAL : DETECTION OF ANTIVIRAL ANTIBODIES IN SERUM USING AN ELISA TECHNIQUE.

Antibody is produced by the body in response to the stimulus. This antibody is specific for a particular antigen and will react with the antigens and neutralize them. The antibodies are a large group of protein molecules present in the serum and tissue fluids. They are also called immunoglobulins (Edwards 1999). Clinical immunologists use the specific technique for the identification of the antibodies present in the human body. Rapid diagnosis of the antigen is performed using the technique called ELISA (Enzyme linked Immune sorbent assay). In ELISA technique, the antiviral antibodies are immobilized on a membrane or on a plastic plate. The test sample is then added to it (Sheehan 1997). If the viral antigen is present in the sample, then the antigen – antibody interaction occurs through immobilization technique. The unbound side of the antigen is then bound with another antiviral antibody. The binding of the second antiviral antibody is measured by using the calorimetric reading. The concentration of the antigen is detected by the intensity of the color formed (Kemeny 1991). Measles, Mumps, Rubella and Cytomegalovirus are serious viral infections. Measles and Mumps belong to the Paramyxoviruses group and they cause infection at the upper respiratory tract (Kemeny 1991). Cytomegalovirus belongs to the herpes viruses’ family and infects the lymphatic system. The last infection is Rubella from the Togaviridae family which causes infection in children. These viruses have the great danger of becoming fatal (Wild 2005). In this experiment, Measles, Mumps, Rubella and Cytomegalovirus viral antigens were immobilized on the micro titer plate and the obtained patient’s sera was added to the wells for immobilization. The plate was then checked for the presence of the particular disease by using the conjugate. The intensity of the color produced by the wells indicates the presence of the disease in the patient. If no color change is observed, it indicates the absence of the disease in the patient (Crowther 2001). Materials: Micro titer plate previously coated with viral antigens 2 x 5 ml Incubation buffer (IB) for diluting patient's serum 0.5 ml 2 patient’s serum samples labeled P1 and P2 8.0 ml Incubation buffer labeled 'conj' for diluting conjugate (made to volume). 2 ml Positive control serum for all 4 viruses labeled '+ve' 2 ml Negative control serum for all 4 viruses labeled '-ve' Blocking buffer (bovine serum albumin) in wash bottle (Buffer) Anti-human IgG-alkaline phosphates’ conjugate (Sigma) 8 ml 1 mg/ ml p-nitrophenyl phosphate in glycine buffer (pH 10.4) labeled 'Substrate' 5 ml 3M NaOH labeled 'Stop' P200 and P20 Automatic pipettes + tips Wad of paper towels positioned next to sink Method: At the outset, the plates were washed well and initials were marked. Following it, 200 µl of diluted viral antigens were added to the wells of the micro titer plates and the plates were incubated at room temperature for thirty minutes and at 4oC overnight. Since only 32 wells were used for the experiment, the remaining wells were blocked by using the blocking buffer. The wells are emptied again and then refilled with the blocking buffer, except 32 wells, and were incubated for three minutes and tipped off. After this process, the plates were drained.