ELISPOT

ELISPOT (Enzyme-linked immunosorbent spot)-a method for detecting cells (T-lymphocytes) that secrete certain products (cytokines, mediators, etc.).

In the diagnosis of borreliosis, along with the determination of specific humoral immunity (antibody tests), the Elispot assay is used to determine the cellular immune response (T-lymphocytes) to Borrelia antigens.

ELISPOT gives both qualitative (type of immune protein) and quantitative information (number of cells activated by a particular antigen).

Antibody tests for Borrelia burgdorferi in the early period (on average up to 3-6 weeks after infection) are not very informative. A sufficiently long serologic window prevents proper diagnosis and treatment, especially in sterile manifestations of stage 1 borreliosis. The production of antibodies in the amounts necessary for diagnosis by existing tests can be disrupted after prolonged use of glucocorticosteroids, cytostatics. Often by antibody tests it is not possible to control the effectiveness of treatment of borreliosis, as after antibiotic treatment antibody titers may persist for a long time, sometimes years. In the case of chronic persistent infection, the low informativeness of antibody tests is due to the small number of the pathogen, the predominance of l-forms of bacteria and disorders in the immune response.

The ELISPOT assay will help eliminate the shortcomings of serologic tests.

The unique high sensitivity of the ELISPOT assay makes it possible to identify even small cell populations in antigen-specific responses. This high sensitivity is largely due to the fact that the detectable product is on the surface of the secreting cell at the time of analysis, bound to its receptors before it dissolves in the liquid phase after centrifugation or degrades. This makes the ELISPOT assay significantly more sensitive than ELISA. Its sensitivity limits are below 1/100,000, making this method very useful for monitoring antigen-specific responses in various areas of immunologic research, including infectious diseases.

Benefits of the Borrelia ELISPOT assay:

• is highly sensitive and can detect even a single T-cell reacting to an infection
• 20 to 200 times more sensitive than conventional ELISA (ELISA)
• suitable for the diagnosis of both fresh infection and chronic borreliosis
• can be useful in monitoring treatment. ELISPOT usually becomes negative about 4 to 8 weeks after completion of effective therapy.

Antigens in the Borrelia ELISPOT assay :

• Borrelia burgdorferi antigen in full: Borrelia burgdorferi B31 reference strain Borrelia burgdorferi sensu stricto
• Borrelia burgdorferi protein combination: OspA Borrelia burgdorferi sensu stricto , Borrelia afzelii, Borrelia garinii + native OspC + recombinant DBPA
• Borrelia burgdorferi LFA-1 (lymphocyte functional protein): the body's own protein + Borrelia burgdorferi sensu stricto (epitope). The indicator is often associated with autoimmune diseases: collagenoses, rheumatoid arthritis, vasculitis.

In the lab. ArminLabs The ELISPOT assay is available for the diagnosis of infections caused by pathogens:

• Borrelia Burgdorferi
• Borrelia Miyamoto
• Anaplasma/Ehrlichia
• Yersinia
• Chlamydia pneumoniae
• Chlamydia trachomatis
• Epstein-Barr virus (EBV)
• Herpes simplex virus types 1 and 2 (HSV 1/2)
• Cytomegalovirus (CMV)