· Used Antigen: combination of selected parts of the specific antigens of Bordetela pertussis (PT, FHA, ACT, TCF), and Bordetella parapertussis (Pertactin, FimN EntA).
· Recombinant antigens are applied in the form of microdots (spots) in triplets to a nitrocellulose membrane in a microtiter well format.
· If specific antibodies are present in the sample, they will bind to the respective antigens.
· The complex is labelled with Conjugate and detected through a colour reaction with substrate (BCIP/NBT).
· The kit enables 48 tests.
· Short incubation periods, total assay time: approximately 1.5 h.
· Each individual test includes control spots, which serves to check functionality and sensitivity of the sets, and also the spots for checking the presence of the conjugate.
· Positive controls included enabling validation of the tests.
· For quantitative evaluation the wells contain calibration spots.
· All reagents supplied are ready to use.
· The kit may be used sequentially for smaller batches of samples (reagents are provided in sufficient quantities).
· Evaluation using Microblot-Array reader and the appropriate software.
· Differential diagnostics of Bordetella pertussis and parapertussis.
· Confirmatory method for ELISA and/or agglutination reaction.
Brief assay procedure:
1. Dilute samples serum/plasma (1:51).
2. Pipette diluted samples.
3. Incubate at room temperature for 30 min.
4. Wash 3 times for 5 min.
5. Add Conjugate.
6. Incubate at room temperature for 30 min.
7. Wash 3 times for 5 min.
8. Add Substrate (BCIP/NPT).
9. Incubate at room temperature for 15 min.
10. Wash 2 times for 5 min in distilled water.
11. Dry and evaluate of test.