Method for classifying the presence or absence of microorganisms in biological media
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A technology of culture medium and microorganisms, applied in the fields of biochemical equipment and methods, determination/inspection of microorganisms, etc.
Active Publication Date: 2017-03-29
COMMISSARIAT A LENERGIE ATOMIQUE ET AUX ENERGIES ALTERNATIVES +1
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Embodiment 1
[0432] Example 1 - Synthesis of nanoporous monoliths with affinity for p-nitrophenol (pNP)
[0433] This example shows a "mixed" basic sol matrix Si(OMe) 4 -NH 2 Synthesis of TEOS. The chosen synthetic protocol was the sol-gel method of nanoporous matrices for the trapping of monocyclic aromatic species shown in application WO2010 / 004225.
[0434] Starting reagent:
[0435] Precursor:
[0436] TMOS (tetramethoxysilane) (=Si(OMe) 4 )
[0437] APTES, Si(C 3 h 6 NH 2 )(OC 2 h 5 ) 3 ((3-Aminopropyl)triethoxysilane)
[0438] Solvent:
[0439] MeOH (methanol)
[0440] Hydrolysis using ultrapure water
[0441] The molar ratio of:
[0442] Alcoholate / Methanol / Water: 1 / 5 / 4
[0443] TMOS / APTES: 0.97 / 0.03
[0444] For approximately 5 mL of 3% NH 2 The ratio of TEOS sol:
[0445] TMOS1.786mL
[0446] MeOH2.43mL
[0447] APTES0.084mL
[0448] Water 0.864mL
[0449] plan:
[0450] In a Pyrex beaker placed in a bath (ethanol and liquid nitrogen) at -25 °C, mi...
Embodiment 2
[0456]In two biological models (i.e., Escherichia coli ATCC11775 (β-glucuronidase positive) and Hafnia alvei ATCC13337 (β-glucuronidase negative)), β-glucuronidase Acidase activity is the target.
[0457] The selected VOC volatile metabolite is p-nitrophenol (pK a =7.15).
[0458] The substrate chosen was that described in Example 1. The matrix converts pNP to pNP - form cumulative.
[0459] The table below details the composition of "MES-pNPG" medium for the production of p-nitrophenol by the activity of β-glucuronidase.
[0460] This medium is called "MES-pNPG" because MES is its pH buffer and pNPG (4-nitrophenyl-β-D-glucuronide) is its enzyme substrate.
[0461] After calibration (3 points: 4, 7 and 10), check the pH: pH equals 6.09.
[0462]
[0463]
[0464] To utilize the predominant acid form of p-nitrophenol, pass through MES (4-morpholineethanesulfonic acid), buffer the medium to pK a -1, which is 6.15.
[0465] 2.1 - Reference control for enzyme activ...
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Abstract
The present invention relates to a method for determining the presence or absence of microorganisms, said method comprising the following steps: 1) providing a container containing a liquid or semi-solid phase containing a biological medium and a gaseous phase , the biological medium can contain living forms of the microorganisms, nutrient elements, and enzyme substrates that are specific to the microorganisms and can be metabolized into at least one VOC metabolite, the gas phase and the liquid phase or semi-solid phase proximity; 2) exposing at least said liquid phase or semi-solid phase to conditions suitable for said microorganisms to metabolize said enzyme substrates into molecules of said VOC metabolites; and 3) by optically converting , determining the presence or absence of said VOC metabolite, characterized in that said VOC metabolite interacts with a nanoporous matrix realized in a separate form from said enzyme substrate, and characterized in that by optical conversion Detection of changes in the optical properties of the substrate indicates that the substrate interacts with the metabolite.
Description
technical field [0001] The present invention relates to a method for determining the presence of at least one generally pathogenic microorganism, especially in a biological sample, preferably in a physiological sample, most preferably in blood. Background technique [0002] Blood culture is the cultivation of blood in or on a nutrient medium to produce bacterial growth. The aim is to amplify the concentration of the pathogenic species in order to detect it in patients presenting with sepsis by granting conditions favorable for its growth. [0003] Blood culture is currently the only method for detecting blood-borne infections, and for microbiology laboratories, this method exhibits approximately one-third of the activity of frequent tests. [0004] Fundamentally, blood cultures are simple detection tests. The manual conventional technique consists in incubating the flasks containing the biological samples to be analyzed in an oven between 35 °C and 37 °C, and then observin...
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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/04
CPCC12Q1/04C12Q2334/30C12Q2334/10C12Q1/18
Inventor 皮埃尔·马库斯马蒂厄·杜波伊罗拉-海琳·吉耶莫蒂-华·特兰-泰
Owner COMMISSARIAT A LENERGIE ATOMIQUE ET AUX ENERGIES ALTERNATIVES
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