EP2111462A2 - Medium for detecting and/or identifying bacteria - Google Patents
Medium for detecting and/or identifying bacteriaInfo
- Publication number
- EP2111462A2 EP2111462A2 EP08762042A EP08762042A EP2111462A2 EP 2111462 A2 EP2111462 A2 EP 2111462A2 EP 08762042 A EP08762042 A EP 08762042A EP 08762042 A EP08762042 A EP 08762042A EP 2111462 A2 EP2111462 A2 EP 2111462A2
- Authority
- EP
- European Patent Office
- Prior art keywords
- beta
- enzyme
- coli
- galactosidase
- chloro
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
- 241000894006 Bacteria Species 0.000 title claims abstract description 7
- 102000004190 Enzymes Human genes 0.000 claims abstract description 89
- 108090000790 Enzymes Proteins 0.000 claims abstract description 89
- 239000000758 substrate Substances 0.000 claims abstract description 60
- 241000588724 Escherichia coli Species 0.000 claims abstract description 52
- 238000001514 detection method Methods 0.000 claims abstract description 41
- 230000000694 effects Effects 0.000 claims abstract description 28
- 238000009654 indole test Methods 0.000 claims abstract description 19
- 238000000034 method Methods 0.000 claims abstract description 19
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 claims abstract description 18
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 claims abstract description 18
- 239000012472 biological sample Substances 0.000 claims abstract description 7
- 102000005936 beta-Galactosidase Human genes 0.000 claims description 22
- 108010005774 beta-Galactosidase Proteins 0.000 claims description 22
- 102000053187 Glucuronidase Human genes 0.000 claims description 20
- 108010060309 Glucuronidase Proteins 0.000 claims description 20
- 239000000411 inducer Substances 0.000 claims description 18
- 102000006995 beta-Glucosidase Human genes 0.000 claims description 17
- 108010047754 beta-Glucosidase Proteins 0.000 claims description 17
- 102000005840 alpha-Galactosidase Human genes 0.000 claims description 16
- 108010030291 alpha-Galactosidase Proteins 0.000 claims description 16
- -1 beta-ribosidase Proteins 0.000 claims description 16
- GUBGYTABKSRVRQ-CUHNMECISA-N D-Cellobiose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-CUHNMECISA-N 0.000 claims description 12
- 108090000915 Aminopeptidases Proteins 0.000 claims description 11
- 102000004400 Aminopeptidases Human genes 0.000 claims description 11
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 claims description 10
- 108090000371 Esterases Proteins 0.000 claims description 7
- QNAYBMKLOCPYGJ-UHFFFAOYSA-N D-alpha-Ala Natural products CC([NH3+])C([O-])=O QNAYBMKLOCPYGJ-UHFFFAOYSA-N 0.000 claims description 5
- QNAYBMKLOCPYGJ-UWTATZPHSA-N L-Alanine Natural products C[C@@H](N)C(O)=O QNAYBMKLOCPYGJ-UWTATZPHSA-N 0.000 claims description 5
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 claims description 5
- 239000004395 L-leucine Substances 0.000 claims description 5
- 235000019454 L-leucine Nutrition 0.000 claims description 5
- 108700019535 Phosphoprotein Phosphatases Proteins 0.000 claims description 5
- 102000045595 Phosphoprotein Phosphatases Human genes 0.000 claims description 5
- 229960003767 alanine Drugs 0.000 claims description 5
- 229960003136 leucine Drugs 0.000 claims description 5
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 claims description 4
- 239000008101 lactose Substances 0.000 claims description 4
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 claims description 3
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 claims description 3
- AEMOLEFTQBMNLQ-AQKNRBDQSA-N D-glucopyranuronic acid Chemical compound OC1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O AEMOLEFTQBMNLQ-AQKNRBDQSA-N 0.000 claims description 3
- 108010000540 Hexosaminidases Proteins 0.000 claims description 3
- 102000002268 Hexosaminidases Human genes 0.000 claims description 3
- 102100024295 Maltase-glucoamylase Human genes 0.000 claims description 3
- 125000003047 N-acetyl group Chemical group 0.000 claims description 3
- 102000004316 Oxidoreductases Human genes 0.000 claims description 3
- 108090000854 Oxidoreductases Proteins 0.000 claims description 3
- 102000016387 Pancreatic elastase Human genes 0.000 claims description 3
- 108010067372 Pancreatic elastase Proteins 0.000 claims description 3
- 229920002472 Starch Polymers 0.000 claims description 3
- 102000005262 Sulfatase Human genes 0.000 claims description 3
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 claims description 3
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 claims description 3
- 108010028144 alpha-Glucosidases Proteins 0.000 claims description 3
- 230000015572 biosynthetic process Effects 0.000 claims description 3
- 239000001913 cellulose Substances 0.000 claims description 3
- 229920002678 cellulose Polymers 0.000 claims description 3
- 229940097042 glucuronate Drugs 0.000 claims description 3
- 229930182480 glucuronide Natural products 0.000 claims description 3
- 150000008134 glucuronides Chemical class 0.000 claims description 3
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 claims description 3
- BOFXVYGDIRCHEQ-GHQVIJFQSA-N methyl beta-D-glucuronoside Chemical compound CO[C@@H]1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O BOFXVYGDIRCHEQ-GHQVIJFQSA-N 0.000 claims description 3
- 239000000049 pigment Substances 0.000 claims description 3
- 239000008107 starch Substances 0.000 claims description 3
- 235000019698 starch Nutrition 0.000 claims description 3
- 108060007951 sulfatase Proteins 0.000 claims description 3
- 238000003786 synthesis reaction Methods 0.000 claims description 3
- 150000008495 β-glucosides Chemical group 0.000 claims description 3
- YBLJYSTXRAWBBH-XUUWZHRGSA-N (2r,3r,4s,5s,6r)-6-(hydroxymethyl)-2-methyloxane-2,3,4,5-tetrol Chemical compound C[C@@]1(O)O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O YBLJYSTXRAWBBH-XUUWZHRGSA-N 0.000 claims description 2
- ZIIUUSVHCHPIQD-UHFFFAOYSA-N 2,4,6-trimethyl-N-[3-(trifluoromethyl)phenyl]benzenesulfonamide Chemical compound CC1=CC(C)=CC(C)=C1S(=O)(=O)NC1=CC=CC(C(F)(F)F)=C1 ZIIUUSVHCHPIQD-UHFFFAOYSA-N 0.000 claims description 2
- 241000644323 Escherichia coli C Species 0.000 claims description 2
- 108010064785 Phospholipases Proteins 0.000 claims description 2
- 102000015439 Phospholipases Human genes 0.000 claims description 2
- 125000001547 cellobiose group Chemical group 0.000 claims description 2
- FYGDTMLNYKFZSV-ZWSAEMDYSA-N cellotriose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@@H](O[C@@H]2[C@H](OC(O)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O FYGDTMLNYKFZSV-ZWSAEMDYSA-N 0.000 claims description 2
- 101710099631 Beta-glucosidase 9 Proteins 0.000 claims 1
- 102100032487 Beta-mannosidase Human genes 0.000 claims 1
- 108010012864 alpha-Mannosidase Proteins 0.000 claims 1
- 102000019199 alpha-Mannosidase Human genes 0.000 claims 1
- 108010055059 beta-Mannosidase Proteins 0.000 claims 1
- 239000006035 Tryptophane Substances 0.000 abstract 1
- 229960004799 tryptophan Drugs 0.000 abstract 1
- 239000002609 medium Substances 0.000 description 31
- SIKJAQJRHWYJAI-UHFFFAOYSA-N Indole Chemical compound C1=CC=C2NC=CC2=C1 SIKJAQJRHWYJAI-UHFFFAOYSA-N 0.000 description 12
- OPIFSICVWOWJMJ-LNNRFACYSA-N 5-bromo-4-chloro-3-indolyl beta-D-glucoside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CNC2=CC=C(Br)C(Cl)=C12 OPIFSICVWOWJMJ-LNNRFACYSA-N 0.000 description 11
- 239000000523 sample Substances 0.000 description 11
- 244000005700 microbiome Species 0.000 description 10
- 150000003839 salts Chemical class 0.000 description 10
- 239000003153 chemical reaction reagent Substances 0.000 description 7
- 230000002255 enzymatic effect Effects 0.000 description 7
- 230000002503 metabolic effect Effects 0.000 description 7
- PZOUSPYUWWUPPK-UHFFFAOYSA-N indole Natural products CC1=CC=CC2=C1C=CN2 PZOUSPYUWWUPPK-UHFFFAOYSA-N 0.000 description 6
- RKJUIXBNRJVNHR-UHFFFAOYSA-N indolenine Natural products C1=CC=C2CC=NC2=C1 RKJUIXBNRJVNHR-UHFFFAOYSA-N 0.000 description 6
- 230000035945 sensitivity Effects 0.000 description 6
- WQKHERPPDYPMNX-UHFFFAOYSA-N 6-chloro-3,4-dihydro-2h-naphthalen-1-one Chemical compound O=C1CCCC2=CC(Cl)=CC=C21 WQKHERPPDYPMNX-UHFFFAOYSA-N 0.000 description 5
- IQUPABOKLQSFBK-UHFFFAOYSA-N 2-nitrophenol Chemical compound OC1=CC=CC=C1[N+]([O-])=O IQUPABOKLQSFBK-UHFFFAOYSA-N 0.000 description 4
- ARQXEQLMMNGFDU-JHZZJYKESA-N 4-methylumbelliferone beta-D-glucuronide Chemical compound C1=CC=2C(C)=CC(=O)OC=2C=C1O[C@@H]1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O ARQXEQLMMNGFDU-JHZZJYKESA-N 0.000 description 4
- OPIFSICVWOWJMJ-AEOCFKNESA-N 5-bromo-4-chloro-3-indolyl beta-D-galactoside Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1OC1=CNC2=CC=C(Br)C(Cl)=C12 OPIFSICVWOWJMJ-AEOCFKNESA-N 0.000 description 4
- UCMIRNVEIXFBKS-UHFFFAOYSA-N beta-alanine Chemical compound NCCC(O)=O UCMIRNVEIXFBKS-UHFFFAOYSA-N 0.000 description 4
- 150000001720 carbohydrates Chemical class 0.000 description 4
- 235000014633 carbohydrates Nutrition 0.000 description 4
- 235000013305 food Nutrition 0.000 description 4
- PCKPVGOLPKLUHR-UHFFFAOYSA-N indoxyl Chemical group C1=CC=C2C(O)=CNC2=C1 PCKPVGOLPKLUHR-UHFFFAOYSA-N 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 238000010899 nucleation Methods 0.000 description 4
- YUDPTGPSBJVHCN-DZQJYWQESA-N 4-methylumbelliferyl beta-D-galactoside Chemical compound C1=CC=2C(C)=CC(=O)OC=2C=C1O[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O YUDPTGPSBJVHCN-DZQJYWQESA-N 0.000 description 3
- 102000035195 Peptidases Human genes 0.000 description 3
- 108091005804 Peptidases Proteins 0.000 description 3
- 102100040653 Tryptophan 2,3-dioxygenase Human genes 0.000 description 3
- 101710136122 Tryptophan 2,3-dioxygenase Proteins 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 229940023064 escherichia coli Drugs 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 238000011534 incubation Methods 0.000 description 3
- 238000002955 isolation Methods 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000003550 marker Substances 0.000 description 3
- 235000019833 protease Nutrition 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- KJCVRFUGPWSIIH-UHFFFAOYSA-N 1-naphthol Chemical compound C1=CC=C2C(O)=CC=CC2=C1 KJCVRFUGPWSIIH-UHFFFAOYSA-N 0.000 description 2
- RUFPHBVGCFYCNW-UHFFFAOYSA-N 1-naphthylamine Chemical compound C1=CC=C2C(N)=CC=CC2=C1 RUFPHBVGCFYCNW-UHFFFAOYSA-N 0.000 description 2
- QHYWQIVTVQAKQF-UHFFFAOYSA-N 3,5-dihydroxy-2-phenylchromen-4-one Chemical compound OC=1C(=O)C=2C(O)=CC=CC=2OC=1C1=CC=CC=C1 QHYWQIVTVQAKQF-UHFFFAOYSA-N 0.000 description 2
- PLIKAWJENQZMHA-UHFFFAOYSA-N 4-aminophenol Chemical compound NC1=CC=C(O)C=C1 PLIKAWJENQZMHA-UHFFFAOYSA-N 0.000 description 2
- RUKJCCIJLIMGEP-ONEGZZNKSA-N 4-dimethylaminocinnamaldehyde Chemical compound CN(C)C1=CC=C(\C=C\C=O)C=C1 RUKJCCIJLIMGEP-ONEGZZNKSA-N 0.000 description 2
- HSHNITRMYYLLCV-UHFFFAOYSA-N 4-methylumbelliferone Chemical compound C1=C(O)C=CC2=C1OC(=O)C=C2C HSHNITRMYYLLCV-UHFFFAOYSA-N 0.000 description 2
- YUDPTGPSBJVHCN-YMILTQATSA-N 4-methylumbelliferyl beta-D-glucoside Chemical compound C1=CC=2C(C)=CC(=O)OC=2C=C1O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O YUDPTGPSBJVHCN-YMILTQATSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000588923 Citrobacter Species 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- DZBUGLKDJFMEHC-UHFFFAOYSA-N acridine Chemical compound C1=CC=CC2=CC3=CC=CC=C3N=C21 DZBUGLKDJFMEHC-UHFFFAOYSA-N 0.000 description 2
- 244000052616 bacterial pathogen Species 0.000 description 2
- 229940000635 beta-alanine Drugs 0.000 description 2
- DLRVVLDZNNYCBX-ZZFZYMBESA-N beta-melibiose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@H](O)O1 DLRVVLDZNNYCBX-ZZFZYMBESA-N 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- HOVAGTYPODGVJG-PZRMXXKTSA-N methyl alpha-D-galactoside Chemical compound CO[C@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O HOVAGTYPODGVJG-PZRMXXKTSA-N 0.000 description 2
- VBEGHXKAFSLLGE-UHFFFAOYSA-N n-phenylnitramide Chemical compound [O-][N+](=O)NC1=CC=CC=C1 VBEGHXKAFSLLGE-UHFFFAOYSA-N 0.000 description 2
- 239000006225 natural substrate Substances 0.000 description 2
- MCJGNVYPOGVAJF-UHFFFAOYSA-N quinolin-8-ol Chemical compound C1=CN=C2C(O)=CC=CC2=C1 MCJGNVYPOGVAJF-UHFFFAOYSA-N 0.000 description 2
- 210000002700 urine Anatomy 0.000 description 2
- ZOTDSZHCQZJZPI-UHFFFAOYSA-N 1-phenylacridin-2-amine Chemical compound NC1=CC=C2N=C3C=CC=CC3=CC2=C1C1=CC=CC=C1 ZOTDSZHCQZJZPI-UHFFFAOYSA-N 0.000 description 1
- JMDJHHPCLNGILP-UHFFFAOYSA-N 10h-phenoxazin-1-amine Chemical compound O1C2=CC=CC=C2NC2=C1C=CC=C2N JMDJHHPCLNGILP-UHFFFAOYSA-N 0.000 description 1
- ZZLQHXCRRMUGQJ-UHFFFAOYSA-N 2'-Hydroxyflavone Natural products OC1=CC=CC=C1C1=CC(=O)C2=CC=CC=C2O1 ZZLQHXCRRMUGQJ-UHFFFAOYSA-N 0.000 description 1
- WZSDWSACAGBYQU-UHFFFAOYSA-N 2-(dimethylamino)-3-phenylprop-2-enal Chemical compound CN(C)C(C=O)=CC1=CC=CC=C1 WZSDWSACAGBYQU-UHFFFAOYSA-N 0.000 description 1
- NZRNYAJFDPBMQD-UHFFFAOYSA-N 2-amino-3,4-dichlorophenol Chemical compound NC1=C(O)C=CC(Cl)=C1Cl NZRNYAJFDPBMQD-UHFFFAOYSA-N 0.000 description 1
- CDAWCLOXVUBKRW-UHFFFAOYSA-N 2-aminophenol Chemical compound NC1=CC=CC=C1O CDAWCLOXVUBKRW-UHFFFAOYSA-N 0.000 description 1
- QWZHDKGQKYEBKK-UHFFFAOYSA-N 3-aminochromen-2-one Chemical compound C1=CC=C2OC(=O)C(N)=CC2=C1 QWZHDKGQKYEBKK-UHFFFAOYSA-N 0.000 description 1
- BTJIUGUIPKRLHP-UHFFFAOYSA-N 4-nitrophenol Chemical compound OC1=CC=C([N+]([O-])=O)C=C1 BTJIUGUIPKRLHP-UHFFFAOYSA-N 0.000 description 1
- CJIJXIFQYOPWTF-UHFFFAOYSA-N 7-hydroxycoumarin Natural products O1C(=O)C=CC2=CC(O)=CC=C21 CJIJXIFQYOPWTF-UHFFFAOYSA-N 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- RGCKGOZRHPZPFP-UHFFFAOYSA-N Alizarin Natural products C1=CC=C2C(=O)C3=C(O)C(O)=CC=C3C(=O)C2=C1 RGCKGOZRHPZPFP-UHFFFAOYSA-N 0.000 description 1
- 108010049990 CD13 Antigens Proteins 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 241000588921 Enterobacteriaceae Species 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- HNDVDQJCIGZPNO-YFKPBYRVSA-N L-histidine Chemical compound OC(=O)[C@@H](N)CC1=CN=CN1 HNDVDQJCIGZPNO-YFKPBYRVSA-N 0.000 description 1
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 1
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 208000033809 Suppuration Diseases 0.000 description 1
- 206010052428 Wound Diseases 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 239000008186 active pharmaceutical agent Substances 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- QNHQEUFMIKRNTB-UHFFFAOYSA-N aesculetin Natural products C1CC(=O)OC2=C1C=C(O)C(O)=C2 QNHQEUFMIKRNTB-UHFFFAOYSA-N 0.000 description 1
- GUAFOGOEJLSQBT-UHFFFAOYSA-N aesculetin dimethyl ether Natural products C1=CC(=O)OC2=C1C=C(OC)C(OC)=C2 GUAFOGOEJLSQBT-UHFFFAOYSA-N 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- HFVAFDPGUJEFBQ-UHFFFAOYSA-M alizarin red S Chemical compound [Na+].O=C1C2=CC=CC=C2C(=O)C2=C1C=C(S([O-])(=O)=O)C(O)=C2O HFVAFDPGUJEFBQ-UHFFFAOYSA-M 0.000 description 1
- 229940024606 amino acid Drugs 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 238000012742 biochemical analysis Methods 0.000 description 1
- 238000010876 biochemical test Methods 0.000 description 1
- 239000013060 biological fluid Substances 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 210000001175 cerebrospinal fluid Anatomy 0.000 description 1
- 235000013351 cheese Nutrition 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- WWAABJGNHFGXSJ-UHFFFAOYSA-N chlorophenol red Chemical compound C1=C(Cl)C(O)=CC=C1C1(C=2C=C(Cl)C(O)=CC=2)C2=CC=CC=C2S(=O)(=O)O1 WWAABJGNHFGXSJ-UHFFFAOYSA-N 0.000 description 1
- 239000003593 chromogenic compound Substances 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
- 238000004040 coloring Methods 0.000 description 1
- 238000012505 colouration Methods 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- GLNDAGDHSLMOKX-UHFFFAOYSA-N coumarin 120 Chemical compound C1=C(N)C=CC2=C1OC(=O)C=C2C GLNDAGDHSLMOKX-UHFFFAOYSA-N 0.000 description 1
- 238000005202 decontamination Methods 0.000 description 1
- 230000003588 decontaminative effect Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- AIUDWMLXCFRVDR-UHFFFAOYSA-N dimethyl 2-(3-ethyl-3-methylpentyl)propanedioate Chemical class CCC(C)(CC)CCC(C(=O)OC)C(=O)OC AIUDWMLXCFRVDR-UHFFFAOYSA-N 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 235000013601 eggs Nutrition 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- ILEDWLMCKZNDJK-UHFFFAOYSA-N esculetin Chemical compound C1=CC(=O)OC2=C1C=C(O)C(O)=C2 ILEDWLMCKZNDJK-UHFFFAOYSA-N 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 210000003608 fece Anatomy 0.000 description 1
- HVQAJTFOCKOKIN-UHFFFAOYSA-N flavonol Chemical compound O1C2=CC=CC=C2C(=O)C(O)=C1C1=CC=CC=C1 HVQAJTFOCKOKIN-UHFFFAOYSA-N 0.000 description 1
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 1
- 235000015203 fruit juice Nutrition 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 239000003349 gelling agent Substances 0.000 description 1
- 210000004392 genitalia Anatomy 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 229930182478 glucoside Natural products 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 239000008268 mayonnaise Substances 0.000 description 1
- 235000010746 mayonnaise Nutrition 0.000 description 1
- 235000012054 meals Nutrition 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 210000001331 nose Anatomy 0.000 description 1
- 230000020477 pH reduction Effects 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 229940066779 peptones Drugs 0.000 description 1
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 1
- 150000003905 phosphatidylinositols Chemical class 0.000 description 1
- 210000002381 plasma Anatomy 0.000 description 1
- 239000012429 reaction media Substances 0.000 description 1
- HSSLDCABUXLXKM-UHFFFAOYSA-N resorufin Chemical compound C1=CC(=O)C=C2OC3=CC(O)=CC=C3N=C21 HSSLDCABUXLXKM-UHFFFAOYSA-N 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000011895 specific detection Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
- ORHBXUUXSCNDEV-UHFFFAOYSA-N umbelliferone Chemical compound C1=CC(=O)OC2=CC(O)=CC=C21 ORHBXUUXSCNDEV-UHFFFAOYSA-N 0.000 description 1
- HFTAFOQKODTIJY-UHFFFAOYSA-N umbelliferone Natural products Cc1cc2C=CC(=O)Oc2cc1OCC=CC(C)(C)O HFTAFOQKODTIJY-UHFFFAOYSA-N 0.000 description 1
- 230000002485 urinary effect Effects 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
- 235000013618 yogurt Nutrition 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
- C12Q1/10—Enterobacteria
Definitions
- the field of the invention is that of microbiological biochemical analysis, and in particular the detection and identification of bacteria.
- Pathogenic bacteria including gram-negative bacilli, such as Enterobacteriaceae, are responsible for many diseases, epidemics, etc.
- the species E. coli ⁇ Escherichia coli) is the most represented aerobic species in the digestive tract. . However, their presence in water is a witness of faecal contamination, and some strains are pathogenic and responsible for peritoneal, biliary, appendicular or genital suppuration.
- E. coli Early and specific detection of E. coli makes it possible to propose a suitable solution, in terms of treatment, of decontamination ...
- This detection can be based notably on the use of detection media comprising particular substrates, specific for a metabolic activity, called target metabolic activity, such as that an enzymatic activity of the bacterium that one wishes to detect: by the choice of substrates, depending on whether there is reaction or not, it is possible to characterize the nature of a microorganism.
- the CPS ID 3 medium bioMerieux
- the invention proposes to solve the problems of the state of the art by presenting a new medium particularly suitable for identifying E. coli bacteria in a fast, inexpensive and easy to implement manner.
- an adapted complementary test allows a fast and easy detection of E. coli. More specifically, the inventors in particular, that an indole test carried out on E. coli which does not express the target metabolic activity makes it possible to increase the sensitivity of the test.
- Biological sample means a clinical sample, taken from a sample of biological fluid, or a food sample, derived from any type of food or an environmental sample such as a surface sample, water sample,
- This sample may thus be liquid or solid and may be mentioned in a nonlimiting manner, a clinical sample of blood, plasma, urine, faeces, nose samples, throats, skin, wounds, cerebrospinal fluid, a food sample of water, drinks such as milk, fruit juice; yogurt, meat, eggs, vegetables, mayonnaise, cheese; fish ..., a food sample from a feed intended for animals, such as in particular a sample from animal meal.
- detection medium is meant a medium comprising all the elements necessary for the survival and / or growth of microorganisms.
- This detection medium can either serve only as a detection medium, or a culture and detection medium.
- the culture of the microorganisms is carried out before seeding, and in the second case, the detection medium also constitutes the culture medium.
- the culture medium according to the invention may contain any other additives, for example: peptones or tissue extracts, one or more growth factors, carbohydrates, one or more selective agents, buffers, one or more gelling agents ...
- This culture medium can be in the form of a liquid, a ready-to-use gel, that is to say ready for seeding in a tube, a bottle, or on a petri dish.
- the detection may be carried out in a liquid medium, strip or other solid support
- substrate any molecule capable of generating directly or indirectly a detectable signal due to an enzymatic or metabolic activity of the microorganism.
- the substrate may in particular be a metabolic substrate, such as a source of carbon or nitrogen, coupled to an indicator producing a coloration in the presence of one of the products of the metabolism.
- the substrate can also be an enzymatic substrate, that is to say a substrate that can be hydrolyzed by an enzyme into a product allowing the direct or indirect detection of a microorganism.
- This substrate may in particular comprise a first specific part of the enzymatic activity to be revealed and a second part serving as a marker, hereinafter referred to as a marker part.
- This marker part can be chromogenic, fluorogenic, luminescent, etc.
- chromogenic substrate well suited to solid supports (filter, agar, electrophoresis gel), mention may be made in particular of substrates based on indoxyl and its derivatives, and substrates based on hydroxyquinoline or esculetin and their derivatives, which allow the detection of osidase and esterase activities. Mention may also be made of nitrophenol and nitroaniline substrates and derivatives, making it possible to detect osidase and esterase activities in the case of substrates based on nitrophenol, and peptidase activities in the case of substrates based on nitroaniline.
- substrates based on naphthol and naphthylamine and their derivatives which make it possible to detect the osidase and esterase activities via naphthol, and the peptidase activities via naphthylamine.
- This substrate may in particular, but in a nonlimiting manner, allow the detection of an enzymatic activity such as the activity of an osidase, peptidase, esterase, etc.
- the enzyme substrate may also be a natural substrate, the product of which hydrolysis is detected directly or indirectly.
- Tryptophan for detecting tryptophanase or desaminase activity
- a cyclic amino acid for detecting a desaminase activity
- Phosphatidyl Inositol for detecting phospholipase activity
- the substrate is preferably selected from indoxyl-based substrates (3-Indoxyl, 5-Bromo-3-indoxyl, 5-Iodo-3-indoxyl, 4-Chloro-3-indoxyl, 5-Bromo- 4-chloro-3-indoxyl, 6-bromo-3-indoxyl, 6-fluoro-3-indoxyl, 5-bromo-4-chloro-N-methyl-3-indoxyl, N-methyl-3-indoxyl, .); based on umbelliferone (4-Methylumbelliferone, Cyclohexenoesculetin, ...); Alizarin based; p-Naphtolbenzene; Nitrophenol-based (ortho-Nitrophenol, para-Nitrophenol, ...); at Aminophenol base (para-Aminophenol, Dichloro-aminophenol, ...); Hydroxyquinoline; Cathecol (Cathecol, Dihydroxyflavone, Hydramino
- the substrates used for the detection of a beta-glucuronidase activity may in particular be 4-methylumbelliferyl-beta-glucuronide, 5-Bromo-4-chloro
- the substrates used for the detection of a beta-galactosidase activity may in particular be 4-methylumbelliferyl-beta-galactoside, 5-Bromo-4-chloro
- the substrates used for the detection of a beta-glucosidase activity may especially be 4-methylumbelliferyl-beta-Glucoside, 5-Bromo-4-chloro-3-indolyl-beta-glucoside, 5-Bromo-6-chloro 3-indolyl-beta-glucoside, 6-chloro-3-indolyl-beta-glucoside, Alizarin-beta-glucoside, Cyclohexenosculetin-beta-Glucoside,
- alpha-galactosidase substrate mention may be made of 4-methylumbelliferyl-alpha-galactoside, 5-Bromo-4-chloro-3-indolyl-alpha-galactoside and 5-Bromo-6-chloro-galactoside.
- 3-indolyl-alpha-galactoside 6-chloro-3-indolyl-alpha-galactoside, Alizarin-alpha-galactoside, Nitrophenyl-alpha-galactoside or their salts.
- enzyme A or C expressed by the majority of E. coli, is meant an enzyme which is expressed by more than 80% of E. coli under given conditions.
- Lactose tryptophanase
- enzyme B not expressed by the majority of E. coli, is meant an enzyme which is expressed by less than 20% of E. coli under given conditions.
- beta-glucosidase N-acetyl hexosaminidase
- esterase sulfatase
- beta-cellobiosidase alpha-glucosidase
- deaminase oxidase
- pigment synthesis beta-Alanine aminopeptidase, elastase.
- indole test is meant a test to detect the production of indole by microorganisms.
- the indole produced in a reaction medium is detected using a reagent, such as Kovacs reagent, dimethylamino cinnamaldehyde.
- inducer is meant a compound inducing an increase in the expression of the targeted metabolic activity, any experimental condition being equal, the metabolic activity is greater when the inducer is at an appropriate concentration than when it is absent or at an inappropriate concentration.
- a glucuronide preferably chosen from Glucuronate, methyl-beta-glucuronide.
- beta-galactosidase a beta-galactoside, preferentially chosen from Lactose, Isopropyl-beta-thio-galactoside
- beta-glucosidase a carbohydrate consisting of a carbohydrate bound in the ⁇ position to glucose or carbohydrate with a ⁇ -glucoside subunit, in particular cellobiose, cellulose, starch, cellotriosis, trehalose. Mention may also be made of Methyl- ⁇ -glucoside, Isopropyl- ⁇ -thio-glucoside, Indoxyl- ⁇ -glucoside or Methyl- ⁇ -thio-glucoside.
- alpha-galactosidase an alpha-galactoside, preferentially chosen from Melibiose, methyl-alpha-galactoside.
- the invention relates to a method for detecting and / or identifying Escherichia coli (E. coli) in a biological sample according to which a) the biological sample capable of containing E. coli is inoculated on a medium.
- detection method comprising tryptophan and a substrate of an enzyme A, expressed by the majority of E. coli to obtain colonies of bacteria b) colonies expressing the activity of the enzyme A are detected, and are identified as being E. coli c) colonies not expressing the activity of the enzyme A were detected, an indole test was performed, and colonies with a positive indole test were identified as E. coli.
- the seeding of the microorganisms can be carried out by all the seeding techniques known to those skilled in the art.
- An incubation step can be carried out at a temperature for which the enzymatic activity that one wishes to detect is optimal, that the person skilled in the art can easily choose according to the enzymatic activity to be detected.
- the detection / identification can be carried out by visual examination, colorimetry or fluorimetry.
- the enzyme A is selected from beta glucuronidase, alpha-galactosidase, beta-ribosidase, phosphatase, L-alanine aminopeptidase, L-Leucine aminopeptidase and beta-galactosidase.
- the enzyme A is beta-glucuronidase.
- the substrate of a beta-glucuronidase activity is chosen from 4-methylumbelliferyl-beta-glucuronide, 5-Bromo-4-chloro-3-indolyl-beta-glucuronide and 5-Bromo-6-chloro-3.
- the enzyme A is beta-galactosidase.
- the substrate of a beta-galactosidase activity is chosen from 4-methylumbelliferyl-beta-galactoside, 5-Bromo-4-chloro-3-indolyl- beta-galactoside, 5-Bromo-6-chloro-3-indolyl-beta-galactoside, 6-chloro-3-indolyl-beta-galactoside, Alizarin-beta-galactoside, Cyclohexenoesculetin-beta-galactoside or their salts, at a concentration preferably between 10 and 1000 mg / l, preferably between 20 and 500 mg / l.
- the substrate of said enzyme A is at a concentration of between
- the tryptophan concentration is equal to or greater than 0.02 g / l, preferably equal to or greater than 0.4 g / l.
- the detection medium further comprises a substrate of an enzyme B, not expressed by the majority of E. coli.
- the enzyme B is chosen from beta-glucosidase, N-acetyl hexosaminidase, esterase, sulfatase, beta-cellobiosidase, alpha-glucosidase, deaminase, oxidase, pigment synthesis, beta- Alanine aminopeptidase, elastase.
- the enzyme B is beta-glucosidase.
- the substrate of a beta-glucosidase activity is chosen from A-
- Methylumbelliferyl-beta-Glucoside 5-Bromo-4-chloro-3-indolyl-beta-glucoside, 5-
- the substrate of said enzyme B is at a concentration of between
- the detection medium further comprises a substrate of an enzyme C, expressed by the majority of E. al.
- the enzyme C is identical to the enzyme A, in which case the substrate must be different.
- beta-galactosidase may be mentioned as enzyme A and C.
- the medium comprises, for example, as substrate for the enzyme A, 4-Methylumbelliferyl-beta-glucuronide and as substrate for the enzyme C, 5-Bromo-6-chloro-3-indolyl-beta-glucuronide.
- enzyme C is different from the enzyme A, beta-galactosidase, for example, may be mentioned as enzyme A, in combination with, as enzyme C, alpha-galactosidase.
- Beta-galactosidase in combination with, as enzyme C, beta-glucuronidase, may also be mentioned as enzyme.
- the enzyme C is selected from beta-glucuronidase, alpha-galactosidase, beta-ribosidase, phosphatase, L-alanine aminopeptidase, L-Leucine aminopeptidase, beta-galactosidase.
- the enzyme C is beta-glucuronidase.
- the substrate of a beta-glucuronidase activity is chosen from 4-methylumbelliferyl-beta-glucuronide, 5-Bromo-4-chloro-3-indolyl-beta-glucuronide, 5-Bromo-6-chloro-3 -indolyl-beta-glucuronide, 6-chloro-3-indolyl-beta-glucuronide, Alizarin-beta-glucuronide, Cyclohexenoesculetin-beta-glucuronide or their salts, at concentrations preferably comprised between 10 and 1000 mg / l.
- the enzyme C is alpha-galactosidase.
- the substrate of an alpha-galactosidase activity is chosen from 4-methylumbelliferyl-alpha-galactoside, 5-Bromo-4-chloro-3-indolyl-alpha-galactoside and 5-Bromo-6-chloro-3.
- the substrate of said enzyme C is at a concentration of between 10 and 1000 mg / l, preferably between 20 and 500 mg / l.
- the enzyme A is beta-galactosidase
- enzyme B is beta-glucosidase
- enzyme C is alpha-galactosidase.
- the substrate of the enzyme A is preferably 5-Bromo-4-chloro-3-indolyl-beta-galactoside, 5-Bromo-6-chloro-3-indolyl-beta-galactoside, 6-chloro-3 -indolyl-beta-galactoside, Alizarin-beta-galactoside, at a concentration of between 10 and 1000 mg / l
- the substrate of the enzyme B is preferably 5-Bromo-4-chloro-3-indolyl-beta-glucoside, 5-Bromo-6-chloro-3-indolyl-beta-glucoside, 6-Chloro-3 indolyl-beta-glucoside, Alizarin-beta-glucoside, at a concentration of between 10 and 1000 mg / l,
- the substrate of the enzyme C is preferably 5-Bromo-4-chloro-3-indolyl-alpha-galactoside, 5-Bromo-6-chloro-3-indolyl-alpha-galactoside, 6-chloro-3- indolyl-alpha-galactoside, Alizarin-alpha-galactoside at a concentration of between 10 and 1000 mg / l,
- the enzyme A is beta-galactosidase
- enzyme B is beta-glucosidase
- enzyme C is beta-glucuronidase
- the substrate of the enzyme A is preferably 5-Bromo-4-chloro-3-indolyl-beta-galactoside, 5-Bromo-6-chloro-3-indolyl-beta-galactoside, 6-chloro-3 -indolyl-beta-galactoside, Alizarin-beta-galactoside, at a concentration of between 10 and 1000 mg / l,
- the substrate of the enzyme B is preferably 5-Bromo-4-chloro-3-indolyl-beta-glucoside, 5-Bromo-6-chloro-3-indolyl-beta-glucoside, 6-Chloro-3 indolyl-beta-glucoside, Alizarin-beta-glucoside, at a concentration of between 10 and 1000 mg / l,
- the substrate of the enzyme C is preferably 5-Bromo-4-chloro-3-indolyl-beta-glucuronide, 5-Bromo-6-chloro-3-indolyl-beta-glucuronide, 6-chloro-3 -indolyl-beta-glucuronide, Alizarin-beta-glucuronide, at a concentration of between 10 and 1000 mg / l
- the detection medium further comprises an inducer of the enzyme A, an inducer of the enzyme B and / or an inducer of the enzyme C.
- the inducer of the enzyme A, B or C is at a concentration of between 100 ng / l and 10 g / l, preferably between 10 mg / l and 3 g / l.
- the inducer of said enzyme A or C is preferably a glucuronide, preferably chosen from Glucuronate,
- Methyl beta- glucuronide Methyl beta- glucuronide
- the inducer of said enzyme A or C is preferably a beta-galactoside, preferably selected from Lactose, Isopropyl-beta-thio-galactoside.
- the inducer of the enzyme B is preferably a beta-glucoside, preferentially chosen from methyl-beta-glucose, cellobiose, cellotriose, trehalose, cellulose, starch.
- the inducer of the enzyme B is Cellobiose, at a concentration preferably between 10 mg / l to 10 g / l
- the inducer of the enzyme C is preferably Melibiose, methyl-alpha-galactoside.
- the invention also relates to a detection medium comprising tryptophan, a substrate of a beta-galactosidase enzyme, a substrate of a beta-glucosidase enzyme, and cellobiose.
- the concentration of tryptophan is equal to or greater than 0.02 g / l, preferably equal to or greater than 0.4 g / l.
- the substrate of a beta-glucosidase activity is chosen from 4-methylumbelliferyl-beta-glucoside, 5-Bromo-4-chloro-3-indolyl-beta-glucoside, and 5-bromo-6-chloro-3.
- the substrate of a beta-galactosidase activity is chosen from 4-methylumbelliferyl-beta-galactoside, 5-Bromo-4-chloro-3-indolyl-beta-galactoside, and 5-Bromo-6-chloro-3 -indolyl-beta-galactoside, 6-chloro-3-indolyl-beta-galactoside, Alizarin-beta-galactoside, Cyclohexenoesculetin-beta-galactoside or their salts, a concentration preferably between 10 and 1000 mg / l, preferably between 20 and 500 mg / l.
- the cellobiose is at a concentration of between 10 mg / l and 10 g / l.
- the detection medium further comprises a substrate of an alpha-galactosidase enzyme.
- alpha-galactosidase substrates examples include 4-methylumbelliferyl-alpha-galactoside, 5-Bromo-4-chloro-3-indolyl-alpha-galactoside and 5-Bromo-6-chloro-3-indolyl.
- the invention also relates to the use of a medium as defined above for detecting E. coli.
- Example 1 Contribution of the Indole test to colorless colonies for the detection of Escherichia coli
- Trypcase Soya Agar (bioMérieux) are added Tryptophan at 0,9g / l, 6-Chloro-3-indolyl- ⁇ -glucuronide at 0.15g / l of 5-Bromo-6-chloro-3-indolyl- ⁇ -galactoside at 0.05 g / l and 5-Bromo-4-chloro-3-indolyl- ⁇ -glucoside at 0.1 g / l.
- This medium is or is not added with Cellobiose at 0.5 g / l. These two media are distributed at a rate of 20 ml per petri dish.
- Microorganisms frequently isolated from urinary samples and from the plaintiff's collection are inoculated on these media by semi-quantitative isolation of 10 ⁇ l of a 0.5 McFarland suspension diluted to 20 e .
- the dishes are incubated at 37 ° C. for 24 hours, then the colonies formed are examined visually. The coloration of these colonies is noted.
- An Indole test is performed on colorless colonies using James's reagent (bioMérieux). The results are shown in Table 2 below:
- Table 2 Contribution of the Indole test to a medium combining 6-chloro-3-indolyl- ⁇ -glucuronide, 5-Bromo-6-chloro-3-indolyl- ⁇ -galactoside and 5-Bromo-4-chloro-3-indolyl - ⁇ -glucoside supplemented or not with Cellobiose on the identification of E. coli
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Analytical Chemistry (AREA)
- Toxicology (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
Claims
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
FR0753150A FR2912425B1 (en) | 2007-02-08 | 2007-02-08 | MEDIUM FOR DETECTION AND / OR IDENTIFICATION OF BACTERIA |
PCT/FR2008/050184 WO2008104680A2 (en) | 2007-02-08 | 2008-02-07 | Medium for detecting and/or identifying bacteria |
Publications (1)
Publication Number | Publication Date |
---|---|
EP2111462A2 true EP2111462A2 (en) | 2009-10-28 |
Family
ID=38458115
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP08762042A Withdrawn EP2111462A2 (en) | 2007-02-08 | 2008-02-07 | Medium for detecting and/or identifying bacteria |
Country Status (7)
Country | Link |
---|---|
US (1) | US8334112B2 (en) |
EP (1) | EP2111462A2 (en) |
JP (1) | JP2010517551A (en) |
CN (1) | CN101631875B (en) |
AU (1) | AU2008220704B2 (en) |
FR (1) | FR2912425B1 (en) |
WO (1) | WO2008104680A2 (en) |
Families Citing this family (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
FR2912423B1 (en) * | 2007-02-08 | 2009-03-20 | Biomerieux Sa | MEDIUM FOR DETECTION AND / OR IDENTIFICATION OF BACTERIA |
JP2011244761A (en) * | 2010-05-28 | 2011-12-08 | Nissui Pharm Co Ltd | Medium for detecting enterobacter sakazaki bacteria |
FR3000501B1 (en) | 2012-12-28 | 2015-08-14 | Biomerieux Sa | MICROORGANISM DETECTION MEDIUM COMPRISING AT LEAST ONE ALKYL (THIO) GLYCOSIDE |
FR3004195B1 (en) | 2013-04-03 | 2017-10-06 | Biomerieux Sa | USE OF AT LEAST ONE CHROMOGENIC AND / OR FLUOROGENIC PHOSPHATASE SUBSTRATE FOR DETECTION AND / OR ENTEROBACTERIUM DETECTION IN A BIOLOGICAL SAMPLE. |
US20160168627A1 (en) | 2013-07-18 | 2016-06-16 | Nestec S.A. | Escherichia coli as a marker for hypertriglyceridemia |
GB201319768D0 (en) | 2013-11-08 | 2013-12-25 | Glycosynth Ltd | Naphthalene derived chromogenic enzyme substrates |
TR201912725A2 (en) * | 2019-08-23 | 2021-03-22 | Pamukkale Ueniversitesi | A HYGIENE DETECTION DEVICE |
CN118360365A (en) * | 2024-04-16 | 2024-07-19 | 麒牛药业(珠海横琴)有限公司 | Reagent for detecting microorganism and application thereof |
Family Cites Families (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CA2027536C (en) * | 1990-05-14 | 1993-02-16 | George Chang | Method for determination of e.coli in water |
US6146840A (en) * | 1994-04-29 | 2000-11-14 | The Regents Of The University Of California | Simultaneous enumeration of E. coli and total coliforms |
US5888760A (en) * | 1997-04-10 | 1999-03-30 | Dade Microscan Inc. | Universal test systems and methods of use thereof for identifying multiple families of microorganisms |
FR2881755B1 (en) * | 2005-02-10 | 2012-11-30 | Biomerieux Sa | MEDIA FOR THE SPECIFIC DETECTION OF RESISTANT MICROORGANISMS |
FR2882370B1 (en) * | 2005-02-22 | 2010-12-03 | Alain Rambach | DETECTION OF A MICROORGANISM STRAIN IN A LIQUID SAMPLE |
-
2007
- 2007-02-08 FR FR0753150A patent/FR2912425B1/en not_active Expired - Fee Related
-
2008
- 2008-02-07 JP JP2009548725A patent/JP2010517551A/en active Pending
- 2008-02-07 EP EP08762042A patent/EP2111462A2/en not_active Withdrawn
- 2008-02-07 WO PCT/FR2008/050184 patent/WO2008104680A2/en active Application Filing
- 2008-02-07 CN CN200880004436.0A patent/CN101631875B/en not_active Expired - Fee Related
- 2008-02-07 AU AU2008220704A patent/AU2008220704B2/en not_active Ceased
- 2008-02-07 US US12/448,895 patent/US8334112B2/en active Active
Non-Patent Citations (1)
Title |
---|
See references of WO2008104680A3 * |
Also Published As
Publication number | Publication date |
---|---|
WO2008104680A3 (en) | 2008-11-06 |
FR2912425B1 (en) | 2012-08-31 |
US20100062467A1 (en) | 2010-03-11 |
US8334112B2 (en) | 2012-12-18 |
CN101631875A (en) | 2010-01-20 |
CN101631875B (en) | 2013-05-15 |
AU2008220704A1 (en) | 2008-09-04 |
AU2008220704B2 (en) | 2013-10-10 |
JP2010517551A (en) | 2010-05-27 |
WO2008104680A2 (en) | 2008-09-04 |
FR2912425A1 (en) | 2008-08-15 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
WO2008104681A2 (en) | Bacteria detection and/or identification medium | |
EP2111462A2 (en) | Medium for detecting and/or identifying bacteria | |
EP2111460B1 (en) | Medium for the detection and/or identification of bacteria | |
JP2008530993A (en) | Detection of microbial strains in liquid samples | |
WO2009092982A2 (en) | Method for detecting and/or identifying clostridium difficile | |
EP2877591B1 (en) | Method of detecting oxa-048 carbapenemase producing bacteria | |
EP3094738B1 (en) | Use of at least one substrate of carboxylesterase and/or triacylglycerol lipase for detecting bacteria of the group bacillus cereus | |
EP2981619B1 (en) | Use of at least one chromogenic and/or fluorogenic phosphatase substrate for the detection and/or enumeration of enterobacteria in a sample | |
WO2007000530A2 (en) | Reaction medium for vibrio bacteria | |
FR3000501A1 (en) | MICROORGANISM DETECTION MEDIUM COMPRISING AT LEAST ONE ALKYL (THIO) GLYCOSIDE | |
EP2205976A2 (en) | Biochemical test for confirming the presence of<i>l. monocytogenes</i> | |
EP2804954A1 (en) | In vitro detection of microorganisms exhibiting azoreductase activity | |
FR2816955A1 (en) | Substrate for identifying pathogenic Listeria, is cleaved by an esterase, other than phosphatidylinositol-specific phospholipase C, that is specific for L. monocytogenes | |
EP2670858B1 (en) | Culture medium for microorganisms including para-aminobenzoic acid as a selective agent |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PUAI | Public reference made under article 153(3) epc to a published international application that has entered the european phase |
Free format text: ORIGINAL CODE: 0009012 |
|
17P | Request for examination filed |
Effective date: 20090717 |
|
AK | Designated contracting states |
Kind code of ref document: A2 Designated state(s): AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MT NL NO PL PT RO SE SI SK TR |
|
17Q | First examination report despatched |
Effective date: 20091209 |
|
RIN1 | Information on inventor provided before grant (corrected) |
Inventor name: MONGET, DANIEL Inventor name: PEYRET, MICHEL Inventor name: ORENGA, SYLVAIN Inventor name: PERRY, JOHN Inventor name: ROGER-DALBERT, CELINE |
|
DAX | Request for extension of the european patent (deleted) | ||
GRAP | Despatch of communication of intention to grant a patent |
Free format text: ORIGINAL CODE: EPIDOSNIGR1 |
|
INTG | Intention to grant announced |
Effective date: 20150519 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN |
|
18D | Application deemed to be withdrawn |
Effective date: 20150930 |