DRG Treponema Pallidum IgM Elisa Kit

PRINCIPLE OF THE TEST 
The DRG Treponema pallidum IgM ELISA Kit is a solid phase enzyme-linked immunosorbent assay (ELISA) This ELISA is using a RF-Sorbent. 
The Rheumatoid factor (RF) is a special autoantibody form. These are autoantibodies, which are directed against the Fc fragment of human IgG. 
The RF autoantibodies are mostly class IgM, but may also be class IgA, IgG or IgE. 
Rheumatoid factors are associated with rheumatoid arthritis. But they can also be detectable in other diseases (e.g. tuberculosis, salmonellosis, syphilis, etc.) and even in healthy individuals. In about 5% of all healthy people, elevated RF values can be found; the titer increases with increasing age. 
The use of anti-human IgG antibodies in the RF-sorbent prevents false positive or false negative results.

Patient samples are diluted with Sample Diluent and additionally incubated with IgG-RF-Sorbent, Microtiter wells as a solid phase are coated with Treponema pallidum antigen. 
Pretreated patient specimens and ready-for-use controls are pipetted into these wells. During incubation Treponema pallidum-specific antibodies of positive specimens and controls are bound to the immobilized antigens. 
After a washing step to remove unbound sample and control material horseradish peroxidase conjugated anti-human IgM antibodies are dispensed into the wells. During a second incubation this anti-IgM conjugate binds specifically to IgM antibodies resulting in the formation of enzyme-linked immune complexes. 
After a second washing step to remove unbound conjugate the immune complexes formed (in case of positive results) are detected by incubation with TMB substrate and development of a blue color. The blue color turns into yellow by stopping the enzymatic indicator reaction with sulfuric acid. 
The intensity of this color is directly proportional to the amount of Treponema pallidum-specific IgM antibody in the patient specimen. Optical density at 450 nm is read using an ELISA microtiter plate reader.

QUALITY CONTROL 
It is recommended to use control samples according to state and federal regulations. The use of control samples is advised to assure the day to day validity of results. 
Use controls at both normal and pathological levels. It is also recommended to make use of national or international Quality Assessment programs in order to ensure the accuracy of the results. 
If the results of the assay do not fit to the established acceptable ranges of control materials patient results should be considered invalid. 
In this case, please check the following technical areas: Pipetting and timing devices; photometer, expiration dates of reagents, storage and incubation conditions, aspiration and washing methods. 
After checking the above mentioned items without finding any error contact your distributor or DRG directly.

LIMITATIONS OF USE 
Bacterial contamination or repeated freeze-thaw cycles of the specimen may affect the optical density. In immunocompromised patients and newborns serological data only have restricted value.

Summary and Explanation 
Spirochetes are motile bacteria with a periplasmatic axial filament. All pathogenic species belong to the family Treponemataceae, which includes the three genera: Treponema, Borrelia, and Leptospira. The Treponema are motile bacteria, 5-15 µm in length and 0.2 µm in width, containing about 10 flexible, undulating, spiral shaped rods. Treponema pallidum, the causative agent of Syphilis, is transmitted by direct contact, usually through sexual intercourse. Syphilis along with Gonorrhoea, Chancroid and Lymphogranuloma venereum, designated as a venereal disease, or VD, is an acute and chronic infectious disease. After an incubation period of 12-30 days, the first symptoms to appear are chancres, soon followed by syphilitic ulcers which then spontaneously disappear in a few weeks. During this first stage (primary syphilis) the Treponema pallidum propagates in related lymph nodes to be distributed to the whole body stream. Three further stages of disease follow which are classified as secondary, tertiary, and quaternary syphilis. 
Treatment with antibiotics at the earliest disease stage and prophylactic measures are ways to prevent epidemics. For this purpose, antenatal and donor blood screenings are mandatory in most of countries around the world.