· Used Antigen: combination of selected parts of the specific antigens of Borrelia sp. (VlsE B. afzelii,B. garinii VlsE, VlsE B. b. Sensu stricto, P83, p58, p41 B. afzelii, p41 B. b. Sensu stricto, p39, OspB, OspA B.afzelii, OspA B.garinii, OspA Bb sensu stricto, OspC B.afzelii, OspC B.garinii, OspCBb sensu stricto, OspC B.spielmanii, OspE Napa and p17), a highly specific antigens Anaplasma phagocytophilum (p44, OmpA and Asp62) and recombinant antigen VCA-p18 for exclusion cross-reactivity with EBV.
· 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 96 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.
· Laboratory test for the detection of Lyme borreliosis, confirmatory test to ELISA.
· Diagnostics of anaplasmosis.
Brief assay procedure:
1. Dilute samples serum/plasma (1:51), cerebrospinal fluid (1:3) and synoval fluid (1:17,5).
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.