Elisa report

1. E nzyme- L inkedI mmuno s orbentA ssay

Applications

Principles

Biochemical technique used mainly in immunology.

first and most basic test to determine if an individual is positive for a selected pathogen, such as HIV.

8 cm x 12 cm plastic plate which contains an 8 x 12 matrix of 96 wells, each of which are about 1 cm high and 0.7 cm in diameter.

Qualitative ELISA

Postive or Negative results

Quantitative ELISA

optical density or fluorescent units of the sample is interpolated into a standard curve, which is typically a serial dilution of the target.

Serum Antibody Concentrations

Detecting potential food allergens

(milk, peanuts, walnuts, almonds and eggs)

Disease outbreaks- tracking the spread

of disease

e.g. HIV, bird flu, common, colds, cholera,

STD etc

Detections of antigens

e.g. pregnancy hormones, drug allergen,

GMO, mad cow disease

Detection of antibodies in blood sample for

past exposure to disease

e.g. Lyme Disease, trichinosis, HIV, bird flu

the antibodies fixed to a solid surface, such as the inner surface of a test tube;

a preparation of the same antibodies coupled to an enzyme. This is one (e.g.,-galactosidase ) that produces a colored product from a colorless substrate.

To be able to determine if a serum sample is positive or negativewhich may help in the diagnosis and treatment of a particular diseaselike HIV and leptospirosis and alike.

To be able to apply the proper methods in achieving an accurateresults.

To be able to know the different reagents, their properties andfunctions.

ELISA for Tracking Disease Outbreaks

A yellow tube and plastic transfer was labeled.

The bodily fluid was then transferred into the tube of another group.The samples were then mixed and after which, the half of the shared sample (750 l)was placed in the group s tube.

The sharing protocol was repeated twice.

The 12-well strip was then labeled.On the first 3 wells, it was labeled as + while for the next 3 it was labeled as - .

The remaining wells were labeled with two of the member s initials.

A fresh pipette tip was used to transfer 50 l of the positive control into the + wells (the same was done for the - wells).

50l of the group s sample was transferred into the the appropriately initialed three wells.

There was a 5 minute waiting period so as to allow the proteins in the samples to bind to the plastic wells.

The microplate strip was then tapped onto the paper towels provided.

The well was then filled with wash buffer.

A 50 l of primary antibody was then transferred into the 12 wells of the microplate strip.

The fluids was then allowed to stand for another five minutes so as to allow the antibody to bind.

Washing of the wells was again performed.

A 50 l of secondary antibody was then transferred into the 12 wells of the microplate strip.

A 5 minute waiting period and washing of the microplate wells was again performed (2x).

A 50l of enzyme substrate was then transferred into the 12 wells of the microplate strip.

The enzyme substrate was let to stand for five minutes.

The results were then observed and recorded.

Hwang Jini

Elisa report

Health & Medicine

Transcript of Elisa report