Syphilis (TPA) IgM ELISA Kit

Sample

serum, plasma

Species Reactivity

Human

Detection Method

sELISA

Intended Use

The Syphilis (TPA) IgM Enzyme Immunoassay Kit provides materials for the qualitative and semiquantitative determination of IgM-class antibodies to Treponema pallidum in human serum and plasma. This assay is intended for research use only.

Contents of Kit

1. Microtiterwells: 12×8 (break apart) strips, 96 wells; Wells coated with Treponema pallidum antigen. (incl. 1 strip holder and 1 cover foil).
2. Sample Diluent: 1 vial, 100 mL, ready to use, colored yellow; pH 7.2 ± 0.2.
3. IgG-RF-Sorbent: 1 vial, 6.5 mL, ready to use, colored yellow; Contains antihuman IgG-class antibody.
4. Pos. Control: 1 vial, 2.0 mL, ready to use; colored yellow, red cap.
5. Neg. Control: 1 vial, 2.0 mL, ready to use; colored yellow, yellow cap.
6. Cut-off Control: 1 vial, 2.0 mL, ready to use; colored yellow, black cap.
7. Enzyme Conjugate: 1 vial, 20 mL, ready to use, colored red, antibody to human IgM conjugated to horseradish peroxidase.
8. Substrate Solution: 1 vial, 14 mL, ready to use, Tetramethylbenzidine (TMB).
9. Stop Solution: 1 vial, 14 mL, ready to use, contains 0.2 mol/l H₂SO₄, Avoid contact with the stop solution. It may cause skin irritations and burns
10. Wash Solution: 1 vial, 30 mL (20× concentrated for 600 mL), pH 6.5 ± 0.1 see "Reagents Preparation".
* Contains non-mercury preservatives

Storage

When stored at 2°C to 8°C unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.
Opened reagents must be stored at 2°C to 8°C. Microtiter wells must be stored at 2°C to 8°C. Once the foil bag has been opened, care should be taken to close it tightly again. Opened kits retain activity for two months if stored as described above.

Precision

Intra-assay
The intra-assay (within-run) precision of the CD Syphilis (TPA) IgM ELISA was determined by 20× measurements of 12 serum samples covering the whole measuring range.

Inter-assay
The inter-assay variation of the CD Syphilis (TPA) IgM ELISA was determined with 3 samples with 2 production kits in 10 independent runs with 2 replicates per run.

Detection Range

The range of the assay is between 0.52 - 60 CD Units/mL.

Sensitivity

The analytical sensitivity of the CD ELISA was calculated by adding 2 standard deviations from the mean of 20 replicate analyses of the negative control and was found to be 0.52 CD Units/mL (OD450 = 0.025).

General Description

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, 5-15μ in length and 0.2μ 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.