Test principle

• > FASTest® / Rapid test

  Lateral Flow Test

  The FASTest® are based on latest rapid immunochromatographic technique. Positive samples contain extracellular antigens or antibodies forming specific antigen-antibody complexes in the conjugate pad area. Migrating (“Lateral Flow”) along the membrane, these antigen-antibody complexes are bound to membrane fixed antibodies or recombinant antigens forming a TEST line.

  A correct test procedure will be indicated by a CONTROL line.

  

   

  

• > MegaFLUO® / IFT

  Indirect Immunofluorescence test

  

  In the indirect immunofluorescence test (IFT), specific antibodies present in the prediluted sera bind to the antigens present on the slide. Non-bound unspecific serum proteins are washed off. Fluorescein-marked FLUO FITC antibodies in the conjugate bind to the antigen-antibody complexes. Evaluation is done with a fluorescence microscope (filter system for FITC) with 400× magnification.

• > MegaELISA® / ELISA

  The Enzyme-linked immunosorbent assay (ELISA) is a common method basing on an enzymatic colour reaction. It is used to detect proteins (e.g. antibodies), viruses or low-molecular compounds (e.g. hormones, inflammation markers, toxins) in the sample.

  For the test, the highly specific antibody-antigen interaction is utilised.

  

  Most used diagnostic formats are the indirect ELISA for the detection of antibodies in the sample and the Sandwich ELISA.

  In the indirect ELISA, the antigen is bound to the plate. This antigen binds the specific antibodies in the sample. In the Sandwich ELISA, an analyte-specific antibody is bound to the plate which binds the analyte in the sample.

  The detection is done in both cases by a specific enzyme marked detection antibody (conjugate). The coupled enzyme changes the colour of the specific substrate into a coloured product.

• > MegaLINE® / Blot

  The LINE immunoassay is a simplified form of the Western Blot. Here, purified antigens/proteins are applied on a carrier membrane. Antibodies of the sample bind to these antigens and can be visualised with an enzyme marked detection antibody (conjugate).

   

  

• > Coagulation

  Coagulation is medicinally understood as the clotting of blood or lymph, but also the coagulation of protein.

  1. Blood clotting
  Blood clotting is an essential process during which the bleedings occurring during injury are stopped. Therefore, the excessive leakage of blood from the blood circulation is prevented and the prerequisites for wound healing are made.

  Coagulation disorders are based on primary and secondary defects or causes. The primary hemostasis is based on cellular factors and can be measured directly in the animal (e.g. SURGICUTT® Vet.), whereas the secondary hemostasis is the “real” blood clotting. Here, the clotting factors play an important role. That’s why the secondary hemostasis can also be determined in vitro, for example with the help of ACT-VETube.

  

  2. Protein coagulation
  “Coagulation” is also known for protein coagulation. This principle is used for example in the examination of punctate from abdominal or pleural effusion of the cat for the detection or exclusion of an ongoing FIP infection by means of the RIVALTA FIP-VETube test.

• > Agglutination

  Agglutination means a visible clumping of antigens and antibodies. The binding of specific antibodies (agglutinins) to defined antigens (agglutinogens) is called active agglutination.

  

  During passive agglutination, at least a third component (normally a second antibody) is participating.

• > Cultivation

  The cultivation of microorganisms on adequate nutrient media is a simple and uncomplicated method for the detection of microorganism as far as growing conditions (e.g. temperature, oxygen, pH value etc.) are known.

• > PCR

  The PCR (polymerase chain reaction) allows the selective multiplication of determined genes from the highly molecular DNA. Thus, minimal amounts of pathogen DNA can be detected even in the initial stage of infection.

  In the first step of PCR, the purified DNA is denaturated by heat. After decrease of the temperature, the specific primers can bind to the DNA (hybridisation). Starting from the primer and in presence of nucleotides, the DNA polymerase synthesises a new DNA strand. This cycle is repeated multiple times to multiply the DNA sequence.

  The detection of the multiplied DNA is done classically by separation on an agarose gel or already in the thermocycler using optical detection methods.

  The iiPCR (insulated isothermal PCR) is a newly developed variation of PCR with simplified extraction and preparation of DNA/RNA and easy-to-read results.