Chlamydia Antigen

Chlamydia Ag ELISA Antigen ELISA is based on the principle of a one step enzyme immunoassay. Urogenital (female endocervical or male urethral) or ophthalmic swabs are obtained from the specimen sample and placed into the Chlamydia Ag ELISA Transport Medium. Urine samples are vortexed for 20 seconds then centrifuged for 20 minutes at 2500 x g and the resulting pellet is resuspended in 1 vial of Chlamydia Ag ELISA Transport Medium. Specimens and controls must be thoroughly mixed, boiled for 10 minutes then cooled before use. Into a pre-treated microwell an aliquot of each of the following are dispensed in the order given – (1) anti-mouse IgG HRP antibody conjugated to horseradish peroxidase enzyme, (2) preboiled specimen or control sample, and (3) mouse IgG monoclonal antibody specific for Chlamydia lipopolysaccharide (LPS). After a 60 min incubation at 37 °C, the wells are washed to remove any unbound material and complexed enzyme is detected by the addition of the chromogenic substrate 3,3′, 5,5′ tetramethylbenzidine (TMB). During incubation, the enzyme reaction produces a blue colour and this reaction is terminated after a specific time with acid which converts any colour produced to an intense yellow. The wells are then read spectrophotometrically. The intensity of the colour reaction is therefore proportional to the amount of Chlamydia antigen in the specimen sample. The mouse monoclonal antibody is genus specific and does not differentiate between C. trachomatis, C. psittaci and C. pneumoniae.