Description:

·         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.

Applications:

·         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.