Catalog # 789061 - 40 tests

Description
Western-blot assay for the qualitative detection of IgG antibodies directed against Borrelia burgdorferi (european strains) in human serum or plasma.

Features
- Based on european strains of Borrelia burgdorferi : B. afzelii "PKO", B. garinii 22 kD Osp C and B.burgdorferi VlsE protein.
- Easy and rapid procedure, realised in 1h30
- Easy reading of results, with the bands location guide provided within the kit
- 2 Positive Controls : strong and weak controls
- 2 Controls of the procedure : sample and conjugate controls

Specimens
Serum or plasma.

Kit Components
Nitrocellulose strips, IgG Positive control, IgG Weak Positive Control, Negative control, Anti-human IgG Enzyme conjugate, Substrate, Sample diluent / Wash buffer, Blocking powder, Band Reading Guide, Record sheets.

Assay Protocol:
1. Wash the appropriate number of strips with 2 ml of diluted Sample diluent / Wash buffer for 5 minutes.
2. Add 80 µl of IgG positive control, 20 µl of IgG weakly reactive control and 20 µl of negative control to the appropriate strips.
3. Add 20 µl of patient samples to the appropriate strips, and incubate at room temperature under agitation for 30 minutes.
4. Eliminate the solution and wash the strips 3 times for 5 minutes with 2 ml of diluted wash buffer.
5. Add 2 ml of freshly diluted IgG enzyme conjugate to each strip, and incubate at room temperature under agitation for 15 minutes.
6. Eliminate the solution and wash the strips 3 times for 5 minutes with 2 ml of diluted wash buffer.
7. Eliminate the solution and wash the strips 1 time for 1 minute with 2 ml of deionised water.
8. Add 2 ml substrate to all strips and incubate untill the 41 kD band of the IgG weakly positive control band appear clearly (4-12 minutes).
9. Stop the reaction by eliminating the substrate, and washing strips 3 times with 2 ml of deionised water.
10. Take the strips out of the plate, and put them on paper to air dry.
11. Compare each band with the IgG positive control and the IgG weak positive control, and register the results.

Interpretation of results
Intensity of bands:
Bands intensity should be compared to the 41 kD band of the IgG weakly reactive positive control.
The bands intensity should be equal or greater than the intensity of the 41 kD band of the weakly reactive positive control.
Interpretative criteria:
- IgG Positive : reaction with 3 or more of the following bands : p14, p17, 22 kD OspC, p30, p39, p43, p58, p100, B. garinii OspC, and reaction with sample and conjugate controls.
- IgG Indeterminate : reaction with 2 of the following bands : p14, p17, 22 kD OspC, p30, p39, p43, p58, p100, B. garinii OspC, and reaction with sample and conjugate controls.-
IgG Negative : reaction with less than 2 of the following bands : p14, p17, 22 kD OspC, p30, p39, p43, p58, p100, B. garinii OspC, and reaction with sample and conjugate controls.
- Unreadable : Strips which exhibit dark staining, spots, streaks or blotches which hinder interpretation.

Storage
The kit must be stored at 2-8°C.

Additional Information

Click here for more information (complete procedure, safety data sheet, literature).