Medium for the Specific Detection of Resistant Microorganisms

Abstract

The invention relates to the use of a combination of two culture media for distinguishing at least three groups of microorganisms in a biological sample, comprising: a first group of microorganisms, belonging to a first taxon of microorganisms and comprising at least a first mechanism of resistance to a first treatment; a second group of microorganisms, belonging to a second taxon of microorganisms and comprising at least a second mechanism of resistance to a second treatment; a third group of microorganisms, that are not resistant to said first and second treatments, said combination of two culture media comprising: a. at least a first substrate for detecting at least a first enzymatic or metabolic activity of said first group of microorganisms; b. at least two markers for differentiating the first group of microorganisms and the second group of microorganisms; c. at least one antimicrobial that is active on said third group of microorganisms.

1 . The use of a combination of two culture media for distinguishing at least three groups of microorganisms in a biological sample, comprising:
 a first group of microorganisms, belonging to a first taxon of microorganisms and comprising at least a first mechanism of resistance to a first treatment;   a second group of microorganisms, belonging to a second taxon of microorganisms and comprising at least a second mechanism of resistance to a second treatment;   a third group of microorganisms, that are not resistant to said first and second treatments,   said combination of two culture media comprising:   a. at least a first substrate for detecting at least a first enzymatic or metabolic activity of said first group of microorganisms;   b. at least two markers for differentiating the first group of microorganisms and the second group of microorganisms;   c. at least one antimicrobial that is active on said third group of microorganisms.   
     
     
         2 . The use of a combination of two culture media as claimed in  claim 1 , said combination comprising at least two markers for differentiating the first group of microorganisms and the second group of microorganisms, at least one of which is an inhibitor of said first or said second mechanism of resistance. 
     
     
         3 . The use of a combination of two culture media as claimed in  claim 1 , said combination comprising at least two markers for differentiating the first group of microorganisms and the second group of microorganisms, which are each an inhibitor of said first or said second mechanism of resistance. 
     
     
         4 . A biplate comprising a combination of two culture media, said combination comprising:
 a) at least a first substrate for detecting at least a first enzymatic or metabolic activity of said first group of microorganisms;   b) at least two markers for differentiating the first group of microorganisms and the second group of microorganisms;   c) at least one antimicrobial that is active on said third group of microorganisms, said antimicrobial being present on each side of the biplate.   
     
     
         5 . A biplate comprising a combination of two culture media, said combination comprising a first medium which comprises:
 a first substrate which makes it possible to identify  E. coli,  chosen from a glucuronidase substrate or a galactosidase substrate;   a first identification marker which is a glucosidase substrate;   a second identification marker which is a resistance inhibitor;   at least one antibiotic;   and a second medium comprising:   a first substrate which makes it possible to identify enterococci, which is a glucosidase substrate;   a first identification marker which is another glucosidase substrate;   at least one antibiotic.   
     
     
         6 . The use of a culture medium for distinguishing at least 3 groups of microorganisms in a biological sample, comprising:
 a first group of microorganisms, belonging to a first taxon of microorganisms and comprising at least one mechanism of resistance to a treatment;   a second group of microorganisms, belonging to a second taxon of microorganisms, different than said first taxon, but comprising at least one mechanism of resistance to a treatment, identical to that of the first group;   a third group of microorganisms, that are not resistant to said treatment,   
       said culture medium comprising:
 at least a first substrate for detecting at least a first enzymatic or metabolic activity of said first group of microorganisms; 
 at least one marker for differentiating the first group of microorganisms and the second group of microorganisms, said marker being a substrate for detecting at least one enzymatic or metabolic activity of said second group of microorganisms; 
 at least one antimicrobial that is active on said third group of microorganisms. 
 
     
     
         7 . A culture medium comprising:
 a first substrate for detecting a beta-glucosidase enzymatic activity,   a second substrate for detecting methyl-alpha-glucoside metabolism, in the presence of a colored indicator,   an antibiotic which is vancomycin.   
     
     
         8 . The use of a medium as claimed in  claim 7 , for distinguishing:
 a first group of enterococcal bacteria developing an acquired resistance to vancomycin;   a second group of enterococcal bacteria developing a natural resistance to vancomycin;   a third group of enterococcal bacteria that are not resistant to vancomycin.   
     
     
         9 . A culture medium comprising:
 a first substrate for detecting a hexosaminidase enzymatic activity,   a second substrate for detecting a beta-glucosidase activity,   an antifungal which is amphotericin B.   
     
     
         10 . The use of the medium as claimed in  claim 9 , for distinguishing:
 a first group of yeasts comprising  Candida albicans  developing a resistance to amphoB;   a second group of yeasts comprising  Candida tropicalis  and/or  C. lusitaniae  and/or  C. kefyr,  developing a resistance to amphoB;   a third group of yeasts that are not resistant to amphoB.   
     
     
         11 . A culture medium comprising:
 a first substrate for detecting a hexosaminidase enzymatic activity,   a second substrate for detecting a phosphatase activity,   an antifungal which is amphoB.   
     
     
         12 . The use of a culture medium as claimed in  claim 11 , for distinguishing:
 a first group of yeasts comprising  Candida albicans  developing a resistance to amphoB;   a second group of yeasts comprising  Candida tropicalis  and/or  C. glabrata  and/or  C. krusei,  developing a resistance to amphoB;   a third group of yeasts that are not resistant to amphoB.   
     
     
         13 . A culture medium comprising:
 a first substrate for detecting a hexosaminidase enzymatic activity;   a second substrate for detecting a beta-glucosidase activity;   an antifungal which is fluconazole.   
     
     
         14 . The use of a culture medium as claimed in  claim 13 , for distinguishing:
 a first group of yeasts comprising  Candida albicans  developing a resistance to fluconazole;   a second group of yeasts, comprising  Candida tropicalis  and/or  C. lusitaniae  and/or  C. kefyr,  developing a resistance to fluconazole;   a third group of yeasts that are not resistant to fluconazole.   
     
     
         15 . A culture medium comprising:
 a first substrate for detecting a hexosaminidase enzymatic activity;   a second substrate for detecting a phosphatase activity;   an antifungal which is fluconazole.   
     
     
         16 . The use of culture media as claimed in  claim 15 , for distinguishing:
 a first group of yeasts comprising  Candida albicans  developing a resistance to fluconazole;   a second group of yeasts, comprising  Candida tropicalis  and/or  C. glabrata  and/or  C. krusei,  developing a resistance to fluconazole;   a third group of yeasts that are not resistant to fluconazole.   
     
     
         17 . A culture medium comprising:
 a first substrate for detecting a beta-glucuronidase or beta-galactosidase enzymatic activity;   a second substrate for detecting a beta-glucosidase or tryptophanase or desaminase activity;   an antibiotic which is ceftazidime.   
     
     
         18 . The use of the medium as claimed in  claim 17 , for distinguishing:
 a first group of  E. coli  ESBL or HL Case bacteria;   a second group of KESC ESBL or HL Case bacteria;   a third group of bacteria that are not resistant to beta-lactamines and/or to cephalosporins.   
     
     
         19 . A culture medium comprising:
 a first substrate for detecting a beta-glucuronidase or beta-galactosidase enzymatic activity;   a second substrate for detecting a beta-glucosidase or tryptophanase or desaminase activity;   an antibiotic which is cefpodoxime;   an antibiotic which is cloxacillin.   
     
     
         20 . The use of a medium as claimed in  claim 19 , for distinguishing:
 a first group of  E. coli  ESBL bacteria;   a second group of KESC ESBL bacteria;   a third group of bacteria that are not resistant to beta-lactamines.   
     
     
         21 . A culture medium comprising:
 a first substrate for detecting a beta-glucuronidase or beta-galactosidase enzymatic activity;   a second substrate for detecting a beta-glucosidase or desaminase or tryptophanase activity;   a combination comprising ceftriaxone and clavulanic acid.   
     
     
         22 . The use of a medium as claimed in  claim 21 , for distinguishing:
 a first group of  E. coli  HL Case bacteria;   a second group of KESC HL Case bacteria;   a third group of bacteria that are not resistant to cephalosporins.   
     
     
         23 . A culture medium comprising:
 a first substrate for detecting a beta-glucuronidase or beta-galactosidase enzymatic activity;   a second substrate for detecting a beta-glucosidase or desaminase or tryptophanase activity;   two antibiotics, which are cefpodoxime and aztreonam.   
     
     
         24 . The use of a culture medium as claimed in  claim 23 , for distinguishing:
 a first group of  E. coli  ESBL or HL Case bacteria;   a second group of KESC ESBL or HL Case bacteria;   a third group of bacteria that are not resistant to beta-lactamines and/or to cephalosporins.   
     
     
         25 . A culture medium comprising:
 at least a first substrate for detecting alpha-glucoside metabolism;   at least a second substrate for detecting a second activity different than alpha-glucoside metabolism;   at least one antibiotic, which is vancomycin.   
     
     
         26 . The culture medium as claimed in  claim 25 , wherein the at least second substrate makes it possible to detect a beta-glucosidase or beta-galactosidase activity. 
     
     
         27 . The culture medium as claimed in  claim 25 , wherein the first substrate is preferably methyl—α glucoside, the second substrate is preferably 5 bromo-4 chloro-3 indolyl-β D glucoside or 6-chloro-3 indolyl-β D glucoside, and the antimicrobial is preferably vancomycin. 
     
     
         28 . The use of a culture medium as claimed in  claim 25 , for distinguishing:
 a first group of vancomycin-resistant microorganisms, comprising  Enterococcus faecalis  and  Enterococcus faecium;      a second group of vancomycin-resistant microorganisms, comprising  Enterococcus casseliflavus  and  Enterococcus gallinarum;      a third group of microorganisms that are not resistant to vancomycin.   
     
     
         29 . The culture medium as claimed in  claim 25 , wherein the first substrate is 5 bromo-4chloro-3 indolyl-N-methyl-a D glucoside or 5 bromo-4 chloro-3 indolyl-a D glucoside, the second substrate is 6-chloro-3 indolyl-β D glucoside or alizarine-β D galactoside or 5 bromo-6 chloro-3 indolyl-β D glucoside or 5 bromo-6 chloro-3 indolyl-β D galactoside or 6 chloro-3 indolyl-β D galactoside, and the antimicrobial is vancomycin. 
     
     
         30 . The use of a culture medium as claimed in  claim 25 , for distinguishing:
 a first group of vancomycin-resistant microorganisms, comprising  Enterococcus faecium;      a second group of vancomycin-resistant microorganisms, comprising  Enterococcus faecalis;      a third group of microorganisms that are not resistant to vancomycin or that express a natural resistance.   
     
     
         31 . The culture medium as claimed in  claim 25 , wherein the first substrate is 5 bromo-4 chloro-3 indolyl-N-methyl-a D glucoside or 5 bromo-4 chloro-3 indolyl-a D glucoside, the second substrate is 6-chloro-3 indolyl-β D glucoside or 5 bromo-6 chloro-3 indolyl-β D glucoside, and the antimicrobial is vancomycin. 
     
     
         32 . The use of a culture medium as claimed in  claim 25 , for distinguishing:
 a first group of vancomycin-resistant microorganisms, comprising  Enterococcus faecalis  and  Enterococcus faecium;      a second group of microorganisms, comprising  Staphylococcus aureus,  that are intermediately resistant or resistant to vancomycin;   a third group of microorganisms that are not resistant to vancomycin.   
     
     
         33 . A culture medium comprising:
 an antibiotic which is cloxacillin;   a resistance mechanism inhibitor, which is a 3rd-generation cephalosporin.   
     
     
         34 . The use of a culture medium as claimed in  claim 28 , for detecting ESBL bacteria. 
     
     
         35 . A culture medium comprising:
 a first substrate for detecting a beta-glucuronidase or beta-galactosidase enzymatic activity;   a second substrate for detecting a beta-glucosidase;   a combination of antibiotics comprising:   cefpodoxime,   cloxacillin,   vancomycin, and   amphoB;   a third substrate for detecting a desaminase activity.   
     
     
         36 . The medium as claimed in  claim 35 , wherein it also comprises a fifth antibiotic which is cefsulodine. 
     
     
         37 . The use of a medium as claimed in  claim 35 , for distinguishing:
 a first group of  E. coli  ESBL bacteria;   a second group of KESC ESBL bacteria;   a third group of bacteria that are not resistant to beta-lactamines and/or to cephalosporins;   a fourth group of Proteeae ESBL bacteria.   
     
     
         38 . The use of a combination of two culture media for distinguishing at least 3 groups of microorganisms in a biological sample, comprising:
 a first group of microorganisms, belonging to a first species of microorganisms and comprising a first mechanism of resistance to a first treatment;   a second group of microorganisms, belonging to a species of microorganisms that is identical to that of said first group of microorganisms, but comprising a second mechanism of resistance to a second treatment, different than that of said first group;   a third group of microorganisms that are not resistant to said first and second treatment;   said combination of two culture media comprising:   a. at least a first substrate for detecting at least a first enzymatic or metabolic activity of said first group of microorganisms;   b. at least one marker for differentiating the first group of microorganisms and the second group of microorganisms, said marker being an inhibitor of the mechanism of resistance to the first treatment and/or to the second treatment;   C. at least one antimicrobial that is active on said third group of microorganisms.   
     
     
         39 . A biplate comprising a combination of two culture media, said combination comprising:
 a. at least a first substrate detecting at least a first enzymatic or metabolic activity of said first group of microorganisms;   b. at least one marker for differentiating the first group of microorganisms and the second group of microorganisms, said marker being an inhibitor of the mechanism of resistance to the first treatment and/or to the second treatment;   C. at least one antimicrobial that is active on said third group of microorganism.