· Antigen used: combination of selected parts of the specific antigens of Bordetella (B. pertussis - pertussis toxin (PT), filamentous hemagglutinin (FHA), adenylate cyclase toxin (ACT), tracheal colonization factor (TCF), B. parapertussis – outer membrane protein (Pertactin), adhesion (FimN), entericidin A (EntA).
· Recombinant antigens are transferred to the nitrocellulose membrane (i.e. to the respective BLOT‑LINE (BL) strips).
· 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 20 tests.
· Short incubation periods, total assay time: approximately 1.5 h.
· High sensitivity and specificity.
· All reagents supplied are ready to use.
· The kit may be used sequentially for smaller batches of samples (reagents are provided in sufficient quantities).
· Positive and Negative controls included enabling validation of the tests.
· BL strips equipped with conjugate control band and a control band indicating kits functionality and sensitivity.
· Colour identification in compliance with antibody classes.
· Identical (i.e. interchangeable) reagents (except for Conjugate) in all BLOT-LINE kits.
· Evaluation of results – the intensity of the bands can be evaluated visually or by means of software (Immunoblot Software).
· Laboratory test for the detection of B. pertusis and B. parapertussis, confirmatory test to ELISA.
· Laboratory test for the detection of B. pertusis and B. parapertussis.
Brief assay procedure:
1. Dilute serum/plasma samples (1:51).
2. Pipette diluted samples and controls into the channels of the tray and insert the BL strips.
3. Incubate for 30 min at room temperature using a shaking apparatus.
4. Wash 3 times for 5 min.
5. Add Conjugate.
6. Incubate for 30 min at room temperature using a shaking apparatus.
7. Wash 3 times for 5 min.
8. Add substrate (BCIP/NBT).
9. Incubate for 15 min at room temperature using a shaking apparatus.
10. Wash 2 times for 5 min in distilled water.
11. Dry the strips and evaluate results using the validation strip enclosed or using Immunoblot Software.