Method for the multiplex serological diagnosis in vitro of spirochete infections

A method for the serological diagnosis, in vitro, of an infection with a spirochete bacterium which is pathogenic in humans, chosen from bacteria of the Borrelia, Leptospira and Treponema genera, by indirect immunodetection, characterized in that the following steps are carried out, comprising:

• 1/bringing a same patient serum sample into contact with
• a/ —a first detection substance and/or respectively second detection substance constituted of an antibody which reacts only with an immunoglobulin of the species of the patient of IgG type and/or respectively IgM type and
• b/ —a plurality of bacterial antigens deposited on several zones of a same solid support, comprising:
• a plurality of bacterial antigens of bacteria, of the Borrelia genus, of different species comprising at least the bacteria Borrelia burgdorferi, Borrelia recurrentis and a tick-borne Borrelia species other than Borrelia burgdorferi, chosen from Borrelia duttonii and Borrelia crocidurae, said bacterial antigens being constituted respectively of at least fractions of bacteria of said species, each said fraction of said bacterium comprising at least one specific antigen and one nonspecific antigen of said species of bacterium, preferably whole bacteria, and
• a plurality of bacterial antigens of bacteria, of the Leptospira genus, of different species comprising at least one commensal species which is not pathogenic in humans and one pathogenic bacterium of the Leptospira interrogans species, said bacterial antigens being constituted respectively of at least fractions of bacteria of said species, each said fraction of said bacterium comprising at least one specific antigen and one nonspecific antigen of said bacterium, preferably whole bacteria, and
• at least one specific bacterial antigen of the bacterium Treponema pallidum, and
• 2/detecting and preferably quantifying the serological reactions of IgG-type and/or IgM-type immunoglobulins present, as appropriate, in said patient serum, which react with at least one of said bacterial antigens, by detecting and preferably quantifying a signal emitted by a ternary complex of an immunological reaction between (a) said bacterial antigen(s), (b) a said immunoglobulin which reacts with at least one of said bacterial antigens, said immunoglobulin being an IgG-type immunoglobulin for detecting and assaying IgG, and/or respectively a said IgM-type immunoglobulin for detecting and assaying IgM, and (c) a said first detection substance for detecting and assaying IgG and/or respectively second detection substance for detecting and assaying IgM.