Catalog # 789071 - 40 tests

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

Features
- Based on european strains of Borrelia burgdorferi : B. afzelii "PKO", and B. garinii 22 kD Osp C
- 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, IgM Positive control, IgM Weak Positive Control, Negative control, Anti-human IgM 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 IgM positive control, 20 µl of IgM 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 IgM 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 22 kD band of the IgM 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 IgM positive control and the IgM weak positive control, and register the results.

Interpretation of results
Intensity of bands:
Bands intensity should be compared to the 22 kD band of the IgM weakly reactive positive control.
The bands intensity should be equal or greater than the intensity of the 22 kD band of the weakly reactive positive control.
Interpretative criteria:
- IgM Positive : reaction with 2 or more of the following bands : p17, p39, p41, B. afzelii (PKO) 22 kD Osp C or B. garinii 22 kD OspC, and reaction with sample and conjugate controls.
- IgM Indeterminate : reaction with 1 of the following bands : p17, p39, p41, B. afzelii (PKO) 22 kD Osp C or B. garinii 22 kD OspC, and reaction with sample and conjugate controls.
- IgM Negative : No significant bands detected.
- 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).