In Vitro Method For the Simultaneous Detection and Identification of Antibiotics of Different Classes and Corresponding Diagnostic Kit

Method for the simultaneous measurement of antibiotics of different classes, comprising:

• (1) bringing a single reactive mixture into contact with a sample to be classified, wherein said reactive mixture is selected from the group consisting of:
• (a) a reactive mixture comprising at least a first marked receptor specific for the recognition of β-lactames, a second marked receptor to specifically and competitively recognize a tetracycline and a biotinylated fragment of a nucleic acid,
• (b) a reactive mixture comprising at least a first marked receptor specific for the recognition of β-lactames, a biotinylated antisulfamide antibody and an analogue of a sulfamide marked either directly or indirectly with colloidal gold,
• (c) a reactive mixture comprising at least one marked receptor that specifically and competitively recognizes a tetracycline and a biotinylated fragment of nucleic acid, as well as an antisulfamide antibody and a marked analogue of a sulfamide, and
• (d) a reactive mixture comprising at least a first marked receptor specific for the recognition of β-lactames, a second marked receptor that specifically and competitively recognizes a tetracycline and a biotinylated fragment of nucleic acid, as well as an antisulfamide antibody and an analogue of a free sulfamide marked either directly or indirectly with colloidal gold;
• (2) incubating the solution obtained in step (1) at a temperature between 30° C. and 50° C. for 3 to 15 minutes;
• (3) immersing a dipstick bearing a recovery system in the solution obtained in step (2) and incubating for 3 to 15 minutes, wherein said recovery system is in the form of:
• a solid support comprising a nitrocellulose membrane to which are attached at two known and distinct sections, called test zones, an antibiotic with a b-lactame core and an avidine, respectively, if said reactive mixture is (a);
• in the form of a solid support comprising a nitrocellulose membrane on which are attached at two known and distinct sections, called test zones, an antibiotic with a β-lactame core and an avidine respectively, if said reactive mixture is (b);
• in the form of a solid support comprising a nitrocellulose membrane to which are attached at two known and distinct sections, called test zones, an antibody specifically targeting either the tetracycline receptor or the specific antibody of the tetracycline receptor and an antibody specifically targeting the antisulfamide antibody, respectively, if said reactive mixture is (c);
• or
• in the form of a solid support comprising a nitrocellulose membrane to which are attached at three known and distinct positions an antibiotic with a β-lactame core, an avidine and an antibody targeting the antisulfamide antibody, respectively, if said reactive mixture is (d);
• and
• (4) interpreting the result on the dipstick either visually by the naked eye or by means of an optical dipstick reader.