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    • 专利摘要:
    The present invention relates to a method for preparing a characterization zone of an analysis plate for carrying out at least one characterization of a population of at least one microorganism in the presence of at least one agent antimicrobial method, by mass spectrometry according to the matrix-assisted desorption ionization mass spectrometry technique, called MALDI, comprising the following successive steps: a step of disposing of an analysis plate for characterization by the MALDI technique, comprising at least one analysis zone carrying an antimicrobial agent, - a step of depositing the population of at least one microorganism on said analysis zone in contact with the antimicrobial agent, - a step of incubation, of keeping the analysis plate, under conditions and for a sufficient time, to allow the interaction of the antimicrobial agent and the microorganism present, - a step of depositing r the analysis zone, a matrix adapted to the MALDI technique; methods of characterization, functionalization, analysis plates and associated uses.
    公开号:FR3024465A1
    申请号:FR1457389
    申请日:2014-07-30
    公开日:2016-02-05
    发明作者:Mahendrasingh Ramjeet；Jeremy Surre
    申请人:Biomerieux SA；
    IPC主号:
    专利说明:

    [0001] The present invention relates to the field of Hibernik Pky. Specifically, nnF relates to the characterization of a podullon minus a micro-organism by mass spectrometry, and to pate, mass spectrometry (MS) by desorption-induced bromination and measurement of flight called MALDI-TOF. The technique can be used to determine the integrity of the microscopy of a careetH cell and is described in the following table: Andramas, in particular, of a microcentrifuge (and made of: MALDI-TOF mass of abundant microorganisms, by comparison with the reference standard, which is used to identify the most complicated and most This procedure often involves mirroring a protocol consisting of a PC, not a single colony colony on a sample of a sample, and a test kit. mass - ent, .. icatIon, by comparison of the baseline data, the baseline was 20% more tect - Iniqu L1a r t was pre - ferred to provide a micro - orc - to e ;, Iota mrrint tetype, espore3e.) the de beta-betere de bs a ddve--ez ... trtles I typ CI CI rïl ', "> 11 says , of type iron..pe /. ,, 25 can be cited the 2012 / 1_3699 n: Lias: At this 2012 / 019G309 2012/023845. The caracterisatlo Jn the eJstance the DETEX,: TIEE lydrolysée of the shape and / or the loss J formc. anttrilot-native that beta-amino acid is incubated with n-L-pnnr; Ethane is used to produce a beta-lectamase. In these patent applications, the preparation of the sample to be dispensed is not described in detail. In general, it is envisaged that the microorganism will be prepared beforehand. ê3, on the plate MAL application VO 2012 / 1'3699, [0041] to [00451). Va ntle5 (Hrabeke, W.:31kova and Kr pen et al., 2012, Studentova, and 2012, al 1-2) give a more detailed analysis of samples to be placed in E, which. following the age-old age, especially the 2011, Hooff, van rbier, Schubert and protocols of the stages of an inocuurn including lonies of 104-4 nism. Centrits' Inn, Cultivated, re-sweated, pelletized with an antit 75L or incubation Per lée min 30 min, 3 hours, 15 cerefugelon, transfer of a m Tcra ture of the superscript of a plate of a micro spectral analyzer LDI-TOF, for which it is necessary to carry out a process according to the invention, As a result, the materials of the consumables can be used in a long-term process in order to achieve the best results in terms of concentration, inoculum and concentration of technology. 1, a 25 cons, nrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrr detection of resistance, given the number of samples per unit of ac- tion between the application of a large number of samples 9: -.) a lack of feedback resuming the identification of resistance, technical MALDI-TOF, and the availability of obtained after the obligatory stage of preparation. Another problem lies in the fact that Ts bacteria are incubated with the antibiotic, in volumes of 10 to 50 μl, which has a significant potential risk of. reduce enzyme-antotic interactions. By a decrease in the size of the cell, the matt Zy. In the case of rtV, it is possible to recognize the robustness of the technique and the robustness of the technic, which are known to produce or secrete low levels of detection. this microbiological agLantir has been described in applications Nos. 20t1, 045544 and US 2012/0: as may be known in the past as MS-MS 1F 1. In this isa analysis mass and.: 1 - .. h` ..: q1C the iOrjSati (.1rr sor sebs on the rne, on the brott. ', 2hioto obtained apt various a, Jitions J rrifIcaLion. In nvention, the irivetiteurs ": to implement a method, a method of one: a cerarization of a micn.) - teichnic organism MAL: return from a resistance to an antin is - put e '7th, r This is 0. The situation is relevant to the catacharacterizations present in this area, of which at least one corresponds to the average of the time and one miter which can correspond to the identity of the microparticle. nisrnic !. The invention also aims to propose a sample containing at least one moiety by ALI: ## STR1 ## rrtletst.o 'if a pop U 0 Fl of one is or not resistant to at least one agent: emps relatively short, of the order of hours Uhr hour, even mcAiis. No, if, in this case, the R & D Area can not be used for the preparation of a R & D program, it would be less expensive to introduce an Artrriembian microarray. said reaction initiating an analysis by mass spectrometry in the course of which the population of at least one microorganism Lcr in the analysis zone is subjected to at least one ionization step; the zone d by a laser mask n the technique of mass spectrometry by pdr maths named NIALF.D: f., .characterized in that qi _rL preparation A rrr nsistant to have a plaqu analysis for a caret, and technical MALD1 corrpr7nrt ru least an area e portant the agent antimicrrje a tantant annose of the population mnin n microorganism on said In this case, the contact zone of a contactor with a contact point Dn, consisting of keeping the same in the past and for a period of time 7, the effect, to allow the antimicrobial agent and the micro-agonist solution to be applied to the zone in accordance with the approved NIALIDL D. technique, In the context of the Ro-Neri pro bono, the nature and nature of the characterization to be characterized is that of the environment, the nature of which is used to characterize it. 't3elori rr: on, the popu. . one of micro -, -.) st st: rsarl, contacting step annannatobien rr, that is to say that the pfx; n, nilatic micro re has iz-ouve iz) an, n-Irn .: its deposition in r "ante with antimicrobial 25 present on itt7 2 :: rare led: LIT; The predestination, which has been evolved, is placed on a population of rr3, ganis e st, obtained after a r concentration of enrichment, riction and cruelty! It corresponds to a colon or a fraction of a cob ~ al. obbEnue after growth on ur uU 30 adapted, including ur medium, t) sé. It is preferably selected from the inventials, cephalides, cephalothiazines, carbapenem-7: monohydrates among ampicillin, ammicillin, acetaminophen, ceftriaxone, cephalaxone, and the like. , fair, year. 3penereal, pitneeini 10 irnipip -ipiperid and faropenem. The preparation is to include a: step: noriotiennalisatior perrnettaiit tFU DtEL: ilr pit d -ei of year 7sur technic, iii-iltDi, including the nei ,, s, -,: ,,, ope por (ese Antimicrobial agent, said forietrannald, is preferably used for the purpose of the invention. The invention also provides for the formation of at least one microparticles. For example, it should be understood that the indication of one or more Wise agents does not include the following: ready-to-use a carrageenan method according to the invention; a process for the reparation of a population of at least one of the following: a microorganism on the characterization, allowing conch, .. Te if a tion of a cro- org-.isme r5.7istant to l'UnnicrobiEm is present on the area of 30 En 1. .., artioilor, analysis pa, r spectrometry mass according to the tec.ue ril-PDF, allowing co n ure if a po 'tion of a rnk: P, 7) -organsme resistant to the antimicroblen agent is present in the characterization zone. In the context of the invention, the antimicrobial age is, for example, divided in such a way as to to allow the idea of resistance due to the production of beta-lactamase, and notammate carbapenemase. The antimicrobial agent is often an antibacterial agent, fertilizer, lime, cobalt, and the like, to control the presence of the antimicrobial agent and / or a product. of degrowth the antimicrobial agent. Loving, the process. characterization according to the invention involves at least two crecrisation, sorting third, the characterization comprises, in addition, ionioncepceon oc tumhe, the genus, or, preferably, the species of pop popui2tîon a pout Preferably, the method of characterization according to the invention comprises: - the identification of the fareTIc, Pu ecre, on the basis of a nisaticui, 'by implementation of a prerequisite, dye spectrometry the area of this cmrometer, mass: MALDI (7- "Ident of a zone Ionization prernWee - careintri_kri :: Ye utiiiinant for the year a pre-calibration - the deti nution of In presence (f.:a.ntuelle corolla none Icterisation u tire iariga of a ncorunicne esncornet anturinuabian, by implementation of a second anaii r -Jr speele [c Tuasse i'z the ALDI-TCE technique corresponds to a shaking zone of character (for Va, i / s (cpux), n Dam; this -; fun microorganism, and deter min nation's eventual stance antimicrobial àl'agent respect of the same microorganism. The invention has to equal; mut puer ci jet a process of - a population of at least u. micro-c.-ganism, the character ... Compounding at least one of the following: The identification of the species of the genus, or, preferably, the species of a population of a microorganism, employs a sample of the same. The MALDI corresponding to a first ionization step of a characterization zone comprising the F fan atian cation of microorganism (s), the antimicrobial agent and MALDI - the determination negates the presence of a population of a microbial resistant microorganism, an antimicrobial agent employing a second MALDI mass spectrometry analysis corresponding to a second ionisau step of a characterization zone comprising. A micro-organism, antirobiotic and a MALDI-adapted method; in that the first and the second analysis are respectively to work a first and a second schooling of a different area of education, including the first one. In the case of at least one anti-organism and anti-antimicrobial agent, the first method uses the first dose of a calibrator to control the cell-mediated urinary tract. It is a matter of course that a single organization organizes, effectively, a preparatory work for the production of the invention. The unspoken, adorn to cret a process I fanctioanalization are a piepuc lysis adapted to the rn. MALUI, channel 7, in that the analysis of the zone of analysis comprises a function zone in the region of which makes the analysis zone carrying a cross-linked agent. Operation can be carried out by depositing an aqueous solution with the agent, followed by drying. Another object mentioned is an assay plate intended to receive a sample of at least one microorganism to be characterized on an area of Elna Inc by mass spectrometry according to the MP- ['I technique, characterized in that what it contains a antimicrobial agent deposited su. The analysis zone of the element 7 is a functionalized analysis zone. In particular, in its plate of the zone (s) of functionalized analysis (s), pc-ause (s) of the antimicrobial agent, has (nt) E as microorganism in one the amount detected by the N-carrier technique (s) or H7 obtained by a telese5 dice and / or the region (s) of 7 functionalized, Cr esu, 7. S 'S lized (s) may be - of a solution solutiot ie poi er, several areas foncuonnaaaaa, any tan Portepjso orientated to a -Hdrobien, with including at least a nremktre area ri e, Fonctk.nnaiL use a first lgent nrUniicroéten, and a second. 7.one 17 .. funcnosnalized, distinct zone and zone, antimicrobial pih_ ade distinct from the - :: :) i-irmriis.r ae ': o iltimicro, n more often-, ed ecra d' s CC n carry c because 0 éo .; (7.7-), ECA is to receive, on the contrary, a pollutant secretion and that: the surface of the zone of emission is bound to become microbial, The use of an analysis plate according to the inversion is shown. The following in-reine: - allows to better understand Vi ion. A schematic top view of a 4 to 4 percent of the mass spectra obtains: Jensp scale, is a relative scale with respect to the intensity of the spse, for example, on a mass range selected from 200 to 1200 nm. z), more intense is 1CP, 1 uM m sanote (as mentioned to the left of the spectra) .The intense peaks will be noted in relation to the most intense peak: a peak intensity of 75 mV will reach cicinc 75 % of Vesheil (on the spectrum containing peak peak intensity of 100 mV) .Therefore the same spectrum, the intensity dls; pcs between the strains ne) had to be run By cons, these spectra perrnette compare, for example, the intensity between the native peak of the robien agent and that of its degradation. as areas of analysis for spot, the sovereign, t fnrrne In order to maximize the safety, it is necessary The face of the plate is electrically conductive, such as a carbon fiber, commercially available in the form of a filter, which is obtained from the surface of the board. of an ECU> ers of one that are not available in Imerne ((disposable). zones of alalysis 1 or spots, t to 20 '' Games or 'believe zones of analysis of reference, which can, in the example shown in FIG. 3rd zone of a population of microorganism (s), matrix of MALDI technique, (1) zone of analysis By the term "oral agent" is meant a protein capable of decreasing, the ability of a drug or antibiotic, to be reduced to one of the following: efention is app e to all micro-orarlsm ... yeasts, molds gold and so, a corresponding antimicrobial agents. Preferably, the agent will be an antibiotic such as an amine, preferably chosen from penicillins, c72, haiosporins, quinines, canines, monobactams and icarcillin. , eeracillin, in particular among Id; c4f + add, liTie, cefuroxime, cero> dinine, -fixime, ceftriaxone, azetonam, eféqi me-néne dnpdnème. in rarer r-d: courses for. 5 gram -5.3UCt (77 0 t; di-aique dor-Kr deposited on the: ':: - o * a oo a uniqe JCI bdc rricro-oro risme p deit est' ne 1,: or other species Gram 15 'does not contain any of the above-mentioned agents, although it is preferred that the agent be antitobrotent to the steel or anti-microbial agent. It is also possible that the chloride type or the necrotic compound may also be exposed to: ## STR1 ## ## STR1 ## the solution should be placed, so that the entire shell should be covered, while the solution is then dried, for example by simply drying the air. and, for example, at room temperature, at room temperature ranging from 17 ° C to 40 ° C, and especially at room temperature. ° C) .It is also the tr-pr sferer ion: a therrnc.statée speaker, for example at 37 ° C, to dry.
    [0002] Very simply, the antimicrobial agent will be deposited in aqueous solution, this deposit being followed by an operation cje. drying. Cr n thus obtains a zone of analysis carrier of raellian, so called forcnalized. it is very likely that the E will be in the area by rays, such as at U, 7 (7) s of Fort. , Nit. Instead, a simple microblank, to be bonded to zone 7, by covalent electrostatic ions, with or without a specific arm or pane (antico Recoilers 10 blotin / Ptireorre, 'pe air', made by the utf32L 2 direracton on the surface of Zmne CrE and r ', adUbiclard, ie, or by Its: 7, w anttil obien and the surface (your deposit will be born, in the same way, in the drawing of a np, a linkage of 9eFier evert binding, which could be used to hide the resistance on ZOne of analysis pern by the person skilled in the art, as a function of the effect of the invention in the context of the invention, above, it is too much of an anti-microbial effect to lay down the noise of the reverse noise, which corresponds to the fact that The intensity or native antirhobar peak could not be observed. rr , 17 rtmrrPrit, compromise the action of be d: a-nases ay. 1! Activity. For example, the 0.04 to 1 g / m2 crageht antirnicrohks. will be deposited. For this purpose, use an ultra-pure solutbri at a concentration of 0.1 to 1% antimicrobial agent. By way of example, for ampicillin, an aqueous solution of 1.7 to 10 ml can be used, and for the faropenem, an aqueous solution of 0.1 to 10 ml can be deposited. 1 mg / ml of faropenem. A characterization zone will be preferred, carrying an antirvcrolDi agent. T also pdr-1,: sitqe. to deposit it in addition to the other compound, noted another antirrrrobial agent of the same family. For example, peninsulas are more preferably carbapenemase. It is therefore possible to use several types of beta-lactam or other beta-lactam reagents in the range of c.72, r2, and etherification. On the other hand, if two anti-icrcdalens are de ned on a ... .77ne zoner semrit ctic !: 77: e way to this! that they should be distinctly different from MALDI-TOF. L ... a7m3 bao, - lactamines, to deposit ornament zori Hblteur Dctama, .. 7th d 15 combriaison L. acid clavubn -! S, in order to csarize me implemented note- In icuhier, we puy- In such a way, it leads to a transition in such a way, in such a way, at least two times more, and has a different antimicrobial effect. It is possible - cBram; cp.i7) ab.DPS MOCSO-20 with one or the same plate. Each (- ,,,, areas will have to be pool: Eause one, antimi, .... n.,) Different bern. THE US SOI CELEFICI.I.1S V EFFECTS.AES IN REPLICA, 'M111 .. antimicm agent will be ssent on areas of analysis., Joire De pli. MALDI toncti as pot rron ',. and therefore 25 provided that will have more E '. Yesterday, then - 'reappointment of, rnatd, re micro-orpr' MALDI. They may be marketed in packaging or in a variety of ways, including the preparation and storage of the micro-organism, which may come from sources. As an example of a source of rr.kr (D e (s), it is possible to clot the samples of non-biological gold, in particular animal or human .This may correspond to a sample of Dicbgî that , whole blood, serum, plasma, urine, liquid cephalo-racaia, tH organic secretion, with u -1 tissue sample or with these isolated cells.This ralivremen, can be deposited such GJA, or will be, of course, sotIrriis Preferably, the zone dan is concerned, a preparation of the type which is obtained at a concentration of at least one extracting step 7, which is known to those skilled in the art. In this case, the population of micro-microscopic microscopy is a product such as VHLC, L c, and any other SC3: D; ej corlarr etrr,: hre p-asmeb, or p. -ceu ') (nrnct) will be 7t !, enric'f-) aa -> (: entry and (1, eXtrUCItiOn r-in the popuiiE: b-Dr! rie rTHoro_3 --- ai, lism (s) to (- '= ask ssve, -this of MiCTO-organn- air.) urra au -able have -ruse culture in b. Hon or ru y an ose to enrich it with an organism. UX, geosed or in lo, are well known from the richness of the world, and are particularly favorable in this respect. t Let Ci COI there. micro-organization: s .., ourircoot "rec:. 't: tre'Jttuas on the 7: Orle C: Dan, - deVint onr.:3 caa' on the 70 ° of analysis, prefer Ti H CeLlaire COrnprt7 nL 'jAtior "1) has (aa, ca no more protesters, in ues r' e 'extfication, In the case of microparticles, the population of microorganisms at least contains microorganisms. As an exerripe, it is possible to deposit 5 to CFU of an organism. It is possible, for example, to deposit a blomam, nutt7, with a phosphoric acid, in water which has been used as a buffer. One of these will be a colony or a reaction of a single species. The deposited population will comprise, in advance, a single species of microorganism, D n ' However, it is not excluded to deposit a population comprising different microorganisms in the analysis zone, in which case it is preferable that the microorganisms are known to be able to develop mechanisms of different resistance. How do I know l, 7: what prescnteraltifi resist 15. was identified. As part of the inseni is not useful to proceed from a sample that would be filed In 4 part is filed, without aribmicr_ well. In z, the population. It can be prepared without a centrifugation step. To do this, you need opulatlon (: 177, micro-detector). You will be able to use the described procedures (..) u. ildentt caPon: HORO-ORGANIZER sting, sena kfs rnar-uiPk, rYuti tKc ..!.: riEa i $ ppo - (: i4 year CO MALDI-7 n conimettlux, such as I e e r e As a result of the Olor Company, Ulieux is no longer able to cajole, in addition to the nodulation of microorganism (s), a known to accelerate reaction in vitro. S Cri C 'of resistance cizorri pretjon rH' ftrc popn'ion of micro-organls 10 rrls, sn contact with vr, an 'iention, -fastidieuse a échanUflor dept.) For example m u: u 25 This cr) In the form of zinc oxide, it is important for the activation of nitro-beta-naphthalenes. It may first be removed from the anion zone in an antimicrobial agent or at any other time in the preparation of the character. The fact that what is deposited in the analysis zone contains a population of a year or a microorganism can be determined by a suitable test in practice, noting that he is born of a colonel solitary on this city. In this way, only one po pu tor of one single processor is nacrato the analysis zone. Incubation of the populajon of microorganism (s) 73 zone - a carrier of both the antimicrobial agent and the infection of a microorganism to be characterized, is subjected to incubation to allow interaction between the antimicrobial micro-ongansm and thus in the case of presence of a microorganism-resistant, anti-microbial host, allow aa reaction / phenomenon causing the resisnce to occur. In particular, when the resistance phenomenon to be detected is in the presence of an enzyme, the reaction will be carried out in such a way as to allow the enzyme reaction to occur. In the case of the invention, the resonant pinenor of antimicrobial agent resonance has therefore been directly on the surface of the microorganism and not at all prior to the deposition. Orgen, Lsm.,. In the presence of this antimicrobial drug, the antimicrobial resistance in the antimicrobial agent occurs in a minute volume of dilution to the zone of anthocyanin, so there is no limitation or inhibition. because previous techniques. The incubation time and incubation time will be laughed and by the skilled person depending on the resistance phenomenon. The slap of ana lysa can be achieved by focusing the range of 17 actens. : Eniptrmaturrs now, especially at bending ground (22 ° C). U is e It may be possible to increase it to a temperature range of 77 ° C, to improve the enzyme response that is important for the CP resistance. Le7. Concluding remarks will be made for drying the micro-ci-c; In particular, when there is a problem to be detected, it involves a hydro-a reaction. The plate may be laced, losing the incubation in a humid atmosphere, unless the moisture content brought directly by the depot containing the microorganism popule is: - In the selected selections .- .. ctionti the incubation time should be sufficient to allow determination of the resistance phenomenon to be detected by MS m_DI-TOF, and the enzyme reaction should be stopped in the case of Dhér '. resistance mediates mediated by an enzyme. Incubation is, as is often done, .erician. 2 ',,, mins 5 minutes, plus., Knent: i at least 2 ilinutes, and even more ellemeenciant a duration, 45 7: 1 90 minutes. He was ,. in the context of the invention, there was no restriction on the time of the 10th organism, it was not necessary to perform such a process, especially in the form of T-TlFe, which consisted of 2-hydroxycinna rrn. Ckh: 2 D.USt. det.tiOn in c technicii, I MALD72: uia population of micro-photosensibli o-dihydroxy isci, 2,: stier, 2 rinamic, nomi-Preux rLO: Tasks CO cxiste ealernent dice the s and acid 2, s. -nnu: -, from Ilorin-7e are now underway. Narrow compo iii perretra cror-M - ,, er les mcilé.cules. ns e c.j2 carat._ isation under the effect of iaser for YLi 25, for consnlJtion this matrix, ur (rIsé is put in solution plus soi..P. ,, - * eni: nce ultra-vire quality Or as an example of a useful solvent, there may be mentioned acetone, acetonitrile, naphthol or ethanol. (TFA) can sometimes be added, for example an example matrix in a water / TFA acetonite at 50 / t / C, and the organic divarit. It is possible to dissolve in water while the hydrophilic solution dissolves.The presence of acid, such as TFA, promotes molecular wrinkles by uptake of L70b. It is then directly deposited on the analysis zone and then covers the microorganism (s) and the arttimicroben agent which are present in the process. in addition, V, 71 nt crionisation ir area antérrrtior.ristallisation step rrr datri, ' This is the case of the matte that is obtained from the ambient air, which is nesent in the nth region, for example, leaving some room for analysis, for example, Drinrne albrit of 1 / nota na ment I'll get it; Ambient C) during this time: K: 5 minutes. This is a case in point where it is urgently necessary for the Republic of Chile to undertake anti-discrimination activities within the United States. It is not subject to any modifications whatsoever: "educe." Radius: rotational erosion, all wavelength irreparable, or even infra rr, Ionisadon may be used, for example, with: a nitrogen laser emitting a LIV radiation at 337.1 (1 m). During ionization, the microorganism (s) and the antimicrobial agent must The matrix absorbs the energy of the photon energy and restores this energy to the sublimation of the disorientation of the particles present in the microarray populaion. if e (s) and t antirni (: reqbien and L ptiarban of matter in a ébi: sma,, air within the r, pro cies e "7" os of charges between n de ar, tl re , micro-atrair in which porA.latlonl micro-! sr.irris, distributes For example, pions may be removed from the matrix and transferred to the proteins, peptides and organic compounds present at the level of the characterization zone. This step allows a gentle ionization of the present molecules without inducing their destn.rfinn. The po. micro-orcianism, $) and the antimicrobial agent Si (Ie), rites iqes, C rniers of which Plus 7.7 (d), and of the acceleration of ion aPP 2.U " The ions go to Itprs see more than the ions plus allowing their separation, at the end of the flight tube, a r-tector. If you do not know how much you are playing, it is the mass spectrum that is obtained, the number of the signal, which is reduced by the reduced volume of the flight tube. pressure tuque and HudenernS of 'under your N eruies is e 20 plus an anérrine TdorrisC1-, 171-11 Once intw Hd, 7-21 rIS the S pectron r, E2iztre a zone décara _ get_, The change in the ratio of the mass ratio to the FAT value of a mass spectrum spectrum of the invention can, therefore, be reduced to the following successive steps. obtaining mass sp.y. - Have a zone to characterize: including the population, the micro-population and the iron ore. 3ger or-link: rober in a spectrometer - possibility to obtain the crystallization of the matrix in which the micro-organized and F-to-rrbien agent are said Ionize the mixture of microorganism anti-microbial-patriotic, thanks to a laser beam, secrete the ionized molecules obteobtenues thanks to the difference of endow let free cpcer I ionized molecules and accelerated in a tube. under pressure reciuittt detect at least a part of the ionized molecules at the output of the tube caror-m'ere me to measure the time q uelies have put to go through the tube under reduced pressure to get corresponding to the nort "-e of molecules tean nt the detector to an instant gold 10 catt :, hard! 71as: on G. manïe L rrup! ecdou3 Lon d ra ept meéc: uk .s Oetectees, In general, calculating the ratio m / z is obtained in tentsm 15 71, Electro-meter, used, hard-on-line on chF: n- - [m / zi and I-park or under-gold; reduction of ionized resins. The calibrator coosses to use a rr o or a MiC ro-op: 1 a Rism a carac., a - s' 1: i -, will provide rnolecu, ot ontseEmi covering a 20 TT d.d _ _._ correspond characterisadca s nsace. This will include the use of StarCards to enable them to measure the rituals of adequate malnutrition. Fortification of the microorganism, ,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,, - [MIZ], depending both on the m / z ratios covering the mass range ut; for ttyo 'cament: in o amrne albnt from 2000 to to tee in, yeast case, i3.ttx corresponding' tttsée from a 30 of I9nsenttesn, you calibrating eeoMIX 6 sserBio L mass from 75n to 1000 ah: ons may, for example, 'naciéries or parasites'. For example, antimicrobial calibration may be peptides of small masses. In c frame covering a range of e used.
    [0003] Te.ut type of spectn -e mass 7-1.A.t., DI-TOF can. to be used for the development of the trap spectrum. Such spectrometers include: an ionization source (generally a UV laser) to ionize the microorganism (s) / crob agent in / matrix; (ii) an accelerator of the ionized molecules by application a large potential of potential; II,) a tube under reduced pressure in which 1e5. e icated e cers; 'y) ii i cC massUnclose the uHares' ormés, in the fioncion of, L.2.dr ratio ciirir rge (mctrei, .. i: asti [rft reL sigrH ecternent by ions mec: 11a par 7,141 DI-TOF is, in the order of 7, to be 5, 5, 5, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 8, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7, 7 p, that more will be counted as a sensitizing factor in the case of an apent andr-nL "Resistance is understood to mean phenorrnene sedor", which means that a substance does not It is not possible to diagnose it via a dimen of its brain exposed to a cancer, but it is reckoned as being on the mass spectrum ob- a given mass peak, or a mass peak change with respect to a mass spectrum of, in particular, with respect to the mass spectrum of the agent anficrls: mb, Dre- At the same time, it has been demonstrated that, in the context of Vinr, it has been shown that a mass spe- on a rrcro <'rrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrr In the presence of an antimicron agent, it was possible to detect the molecules of interest relevant for the expression of at least resistance. resistance, 7th. .., [ttroulation of a cro-organism may dt.o.lc, understand the éit2pes' these al) r .. ug., dare a spectrum this mass of r for example POU -1t: a ntimicro P u pcu Degradation products, such products of L d., 1, leads to resisbricres and, in particular, to the presence of a biodegradable enzyme, such as microparticles. rne (s) and the agent. antirr robien. on the area of analysis t put in the presence of the matrix of a 2 :: -. jr2 of characterization according to nventionu [ste ni i ': cl) in the resistance,: dl) compaer + e sptre de nn, (cl) obtained with cloned stitches and glimpsed with or without a micronized antimicrobial pest, where or if. one or more mass. Characterization of an arachnic irritation is caused by the antitinal agent Ji in. r can be realized, by e, from the ratio utees e n - a nic e mass character :. I, you of the agent ant mbiobien or one of his p. of an uracteristic mass of an external calibrator or from the intensity ratio between a characteristic mass peak of the anti-microbial agent and a mass peak c73N3ct.eristic of a In the case of resistance resistance, a calibration is in a mass range corresponding to low masses, o. the mass of the substance is added after this ionisabon, and the mass is added, to the extent of the presence of the clea, the peak (s) of the mass product plus the products pre-treated, it (s) , we can deduce. Typically, the conversion is in the range of from about ZOO to about 12,000, in the range of from about 200 to about 600 Da. (4: No. of r.-1, -se obte: 1) is also con-s in this mass mne. To realize this calibrati 2 microirtres of a cernante solution composed of a mixture of peptides r LaserBio Labs) and. HCCA matrix, the EO-cyano-4- acid may, exe.mple, be deposited on an area 10 will serve alon: I 5bndards so masses di Le pru for (estectu rs a CPPstano of year / 3e e LrEj'ce. It does not take into account the number of zones in the field of measurement (7_01e par., 7 n é-cules Irrns of the car'a nt.) The permletre can be used in particular. capacity of a micro-organism with a characteristic, and, in particular, with the properties of the micro-organism, the -L .--> Jospp: Drinases s cin.:-.,::,:5s', ,, s es because L is aP, PP! , i dPutups, (yue n, .... '.' ':: - .. a, lsases su r r': i. t 1 0 nur 'mcallscation Enzymatic entp, P sint U ncccccnaller-AlltinliCrObin, for example, include repranisms ,, i.son such as r ag: lotion macrt par, It is a fosfomy for the erection of PYPPs from the Mediterranean and also of the plosplora esamids, acrolides, cleins and aerosols. tetraCyC not, 25 iraclen ti: stop of the rrc: rrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrr These are used with pre-functionalized plates II, which are commercially available. Only caiibration and interpretation will have to be done to allow the detection of antimicrobial resistance in particular to a given antibiotic, of which 3 per cent. routine rapid determination can be avoided in many clinical cases, is now in the present invention, the characterization method according to the invention, the presence of a micro The antibacterial organism, in a very short time, presents a very special case for carrying out a fast (Lai, this is particularly the case for the treatment of Enteroct 75 carbapenemase (CPE), the 0 nc In addition to the above-mentioned criteria, whether or not they have been encountered, there is a correlation between the following: whether or not they are identi -nin-o-organisms, ## EQU1 ##, ## EQU1 ##, ## EQU1 ##, ## EQU1 ## where ## EQU1 ## positive, we can V those of genr es: Enterococcus, 30 .s / 5: e / ria nos-. Spectra of BTEN, by N'IDDI, for such micro-forms, and their major V viSHs, are available and are shown in FIGS. , -, -,,,,,,,,,,,,,,,,,,,,,,,,,,,,. -7 '' '' '' '' '' '' '', '' '' '' '' '' '' '' '' '' '' '' '' '' '' '' '' '' '' '' '' '' '' '' '' '' '' '' '' '' '' ' They are stored in databases (., J) with commercial MALDI-TOF devices, and, by comparison, allow the presence of such microorganisms to be identified. Thus, a micro-organism may be able to understand the following steps: (a2) a reference mass spectrum, Hour at least one homo-organism, and most often for a micro-emission streak anti-microbiological agent in the analysis zone of a: diene the characterization of the material, nπtranantant, to a lonsate (c2) to obtain a spectra of an eyebrow to be cracked in the presence of nistron , of the mHnnnrrrrt and in particular djmfl, e, cp-, or, of rem: n. Ve - ..: e2. of at least one micro-organism. Dark: càs of the identification of microorganisms, a number of microorganisms. The number of raids, Mea In nq ni, L Ci a n s 201000 Da 'and this re cei C 31-1711ffiC ah70 e n to 17000 Da. The spectrum of the ma- ter, "D attained in gmpr: (in this step of Tr) is carried out by placing the pod of a -ice in a zone For example, for this caliDration, use of micro-organism, such as micro-organism, can be used to enforce the procedures for the establishment of early identification of microorganisms, in Tianu The use of the equipment MALDI-TOF comrnercnux, that the apparatus VITEK® MS commertlatized by the soté LHrieux (lH: pec2) 2) is ertaHt the micro-organilar calildrati present on, pie and, cn proceeding his analysis by NIALDI-TOF, such is the case for the zones of the anary, this reference for the cation: one for the identification of the microorganism and another for the identification of the microorganism and another For the characterization of the resistance, it is possible to use the frequency zone for the two calibrations, in which case the reference zone will be fonclionalized with the antimicrobial agent. for which the solution is to be studied. In the context of the invention, there will be mentioned, but not mandatory, taro-it and idenifiCatOrl P will be implemented, or the resistance rreurterlenrre that neE.7.7) -tion and d) Anatomy of this type, of sercur -realty on the zone: (FicEtic ,, 1 u, r, sme. sabons zorrc atic are men ans enle ,,, / E: ,,,, r plate ciVE.:1u Ntk .. It should therefore be assumed that there is less than 1 micro-organism and that there is a series of micro-organisms as well as a spe Ct: 2'2e e of reference 2i., For rager r. eintimicrobien errls for .cluisrs ci; e iercai,. r.nase thanks to .7 ', 7,311orIlt. for Vicia.ritifbdtion and who has been upgraded or referred to above., c3) to screen the population of the organism (s) and agent 25 discharged from the area of exposure. in the presence of the matrix. (corresponding to the characterization method according to the invention), to an ionization, d3) to acquire a mass spectrum o: btenu following this ion7 in the range of Ma77.7 . of interest - the identification of the organism, when those secreted by a microroon 10 will be rifted, neces- sarily characteristic; L. eff. 3, 15. E., Perform a second calibration of the E mass spectrometer using CalICANT 3 for the detection of the resistance, and then go to a fixed area, reposition the poo; atton of micro-gold: C anti-antimicrobial deposited in the analysis zone and put in the presence of the ma tri taa., corresponding to n zone of character -count.rvantion) 'an ionization, Jn be nr7: 35e frafte this ior -se, om de rasse e, - termination r la S e nnpe d3 d Ver - OR - rice mile, the genre, or, from there a microphone preselt. DO -hold J g3) e 10 sistarce h3) cc inc 15 - -erp: -: e 2 and deduce it to reser cc or not oi ) Can be enes at the end of the n-èsnhtion cc.nce up way PI nopulotion square-by-laugh at a 20 pop. the ion of a single microcircle and k reristi 0 n s ent. '..., tion of a rntcro, contser. The same microstructure makes it possible to infe - o from the same zone of earactr, and thus to characterize both the characterization of One of them was resistant to microstructure, so-called, L.-TOF, without a long and tedious pre-treatment of a sample prior to deposition. The characterization of a phenomenon of resistance to the agent is shown by the following example: an IC oppressed directly on the microcargeor cell and by antiincrobial agent, having a posi in some cases, on the detrimental effect of the detection of ice phenol, - a reduction of the po and the cost in terms of manual consumption and for the characterization of a stance phenomenon to an antimicrobial agen; The traceability of the samples is greater than that of any other commercially available antimicrobial agent in the world. to illustrate the invention but n no character read with the apparatus s c. - The analysis of each of the 15 studies was carried out on all the 3 years, and this was done in the light of the calculations. VITEK 1S de lddata / 2.0 , conn: snurr, clans ctt-a Tdent Versron 2.1.0, VItTEUD rytn cornntnercn sition. Vs 20 - for yse s nics ne: i. of the V2.8, were conducted to check only 1 per MA.CU. ... "The ptibiotlque of t'p. ....... . At the end of the day, it was decided to use a MALDI plate. , quoted from a cost-effective solution to a problem in the present experiment, the area of D na'ily, Se d. t MS disposable slides were treated with a beta-laci.anine anabiut and tested to assess the ability of the annbln to be beta-lactamase.Amicillin was utiik.A4, as mcLiud The experiment was carried out by carrying out the following steps: 2 μl of a 10% solution of ampicillin in water, deposited in droplet form, over an area The plate was incubated at 37 ° C., until the droplet became dry, 2-fold, 7-lactate, recombinant (3-lactam). A-ldrich lot L6170-550UN. CS- S: -airotarnase re-binding to Imgimi in water, diluted with water diluted with EC zinc, 10 F, Zinc (O'5.) EnzymatuMy depositing a large area of Halyse Va rjotraraa 15: riceyme eaL with 'ra 3 on the lys zone, a3h.is "carrier-rrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrr Roxy aryl radicals (VITEK trice CHCA,.), wherein: ## STR1 ## - A rrnsc PT and Para, using one in Las e Bic 'C 25 ch "has been S reed' year 47errarone, for this libration of apparelage. Poar rr 7sses. In this case, a dilute (10X) to HCG-1 (Sc) diluent (10X) was repaired using diluent (thiophosphoric acid). , 01% in ultrapure water), according to the recor-labeling of the supplier The peaks were acquired at a range of 200 to 1200 Da and the presence of molecules corresponding to the fast molecule. or its by-products of degradation is studied, all in sata on the plate, shows mass specs obtained and shows that ampicillim is not stable on the plate. and that EPe is effer erly degraded beta-lactar, ase after incub 7. Hpn, Indeed, on L o bserve a majority peak 368.09 z corresponding to the form mcno5, ine co mar :, da nt a complete loss of t3 form 10 fla; ^. / 2 to 372,09 (sr form 7- The experiment was conducted to link the iron deficiency by a variety of bacterial scrapes, from colonies to saline zones, and to a solutainl last one. Lastly, and drying, as described above, the bacterial strains and their characteristics are presented in 1 below. 15 spontaneous: Ince.st: erlF con, r-con'i7sps, ndant corrcn1FZO e, na co nt r51, corresr, rns n atr ,, rt: r marc-. on the top of the artamase. F1.control, for the 09 m / z, this cii picU Ur, »action etszyrrrata. ', By b' Lech: liq '' MALL: 1) 1. by repairing the zone of character According to: YS 20 rd _ _he cL Species MIC * of ampicillin Phenotype Bacterium / virulence factor ** Reference number 1 K. pneumoniae 32 resistant 2 E. neck> 128 resistant enicillinase 3 E. neck> 128 resistant TEM 6-lactamase 4 E. neck 4 not sensitive 6-ATCC 25922 lactamase * MIC: minimal inhibitory concentration of the strain ** Virulence factor: corresponds to the known secreted enzyme The esseis were carried out using the following steps: jo7 10% aqueous polyacrylate (db, 76 mM) was added to the zones of the plate as in Example 1). The pI, incubated at 37 ° C until the solution was completely dry. nitro-organismification (Incubation at 70 ° C to 70 ° C) in one hour in one hour at a time of 70 minutes. The closed container containing water was used as a plate incubator, and 1 μl of an HCCA layer was added to the sample plates. ,,, carrying both arti fi c and bacteria. A MALDI-TOF mass spectrum analysis was performed with ViiEK® NIS equipment, with a vacuum of 13 microliters, once for each SC (b) of a bacterium. cr, 7'-issariu7 ,. ' for 24 hours on a medium Fplica -;; i on areas hurlalySe carrying bter cernm fnd in the process VITE ..) (Jur 31: lacéa in the chamber of the apparatus, leading two steps below: o a first ei: uiit on the spectrum of the characterization zones, after: ation c to start from a reference zone with an E. coli (E-cal) satdiss, a second 7-cc. With the same areas of characterization, there was a 10-year-long test of Ls of Neon, which was followed by a peak of 15 days. After this, after the first time, we should be aware of the fact that we have not included any of the following items in the form of the native form or all the samples have been collected. The analgesic structure of the anastomosis on the earth or the mass of the site: a second series and a smaller series of 20 arripic iron strains. ampicillin was inhibited in the case of strain E, coli ATCC 25922, known to be variable in size. In fact, the second acquisition of the weak masies, a larger area at 372.15 m / z, corre- erably decreases. In the case of ampicillin-resistant strains, the highest peak was 5% to 3%. In the case of ampicillin-resistant strains, the maximum peak was 5% (515 ml), corresponding to the cyclopyran form of Varnpof Mine. ParsianIsparitIon e. As the first acquisition and the observation of the corresponding spectra, all strains of bacteria should be detected: a high confidence level. quaucirre -,: jstarite., ..; As shown in Table 2 below, a correct identification of the bacteria by MALDI-TOF was obtained from the first series of acquisitions for all characterization zones, 1, with a confidence level varying from 99.9 to 100%, showing that the perimenal conditions still make it possible to discriminate the different species Table 2 Strain of Number of Species Probabi bacterium, peaks used after Reference No. by comparison identification Spectra of reference pneumoniae 99.9 pneumoniae 100 95 coli 100 2 103 E. coti 100 91 E. coit 99.9 101 E. cati 99.9 4 94 99.9 4 112 99.9 Example 3. Further tests were carried out to demonstrate that identification and detection of carbapenemase production from the same characterization zone was possible. antibiotic model carbapenem, and a protocol e identical to that used in Example 2 was used. A solution of faropenem was prepared with a concentration of faropenem of 0.5 mg / ml. The bacterial strains used are detailed in Table 3 below.
    [0004] Table 3 Phenotype Species Strain Bacteria / N virulence reference factor S. marcescens resistant 1MP-1 carbapenon-6 E. coli Not sensitive to the presence of ATCC 25922 lactamase in the second place, the mass spectra obtained from the tests and shows that the acida carbapenemase was able to relaunch these studies with the second series of doses. .1 that the forms of faropenem are not affected in this experiment, it has been possible to demonstrate the loss of the two native forms of faropenem, due to the oroduc-Aiiin by the carbapenemiasis bacteria.
    [0005] 10 After the acquisition in Ma reibles. in C h e serisie a majority peak at 308,13 lie and an iniquitic peak miz tere cre fer. eueusponrie nt, Tuve men, two Fa natmes (read: E; e30 nt e 15 Ri .bapér.In short .LJiSNt1L 'ciriliptentinn spectra, Ma Oeux Eiryud-ids correct' d with ri i'ead confidence.Menden .. E-Hrffiiste O VexemplerKl.e- of the zone C uractércerOon idencation si C1: 7; .UU 20 the species by NF type E. col! Of the truOrudata series obtura r: with the se ry discret rs marcescens bacteria, as shown by the ALMn series of accumulators, found in Table 4 K.Doff, GP, J. 3. van K-Dn-Ip.n. R. 3. Meeer A. va.i a) 'Ceessens nJ TM Lu Ider 1.2). "Activation of the enzyme actvt n bacteria ^ ysates using MALDI-rnass spectrorn Prot. 79-84. Hrebek. J., V. Studertova, R. V, p., A., H. Zemlickova, V.: 2akubu E. Chuc Vcreva, M. Gnd: vski, Y Pfefer, J. D. Per y, nso n 1 T.
    [0006] ## EQU1 ## and ## EQU1 ## where: ## STR1 ## and ## EQU1 ## Laser: ## EQU1 ## The strain of Number of peaks Species obability after comparison of reference spectra bectene, used pnr No. reference identification 100 S. marcescens 99.9 5 113 marcescens 99.9 6 109 neck 99.9 6 101 E. colt 99.9 Walkova, V, 5 -tui'iulwv ,,, (. '.. 1-1 Ci C OVe r ri e se ttt: Tecton my tUenea..jst .--' Za 0 r .: e and U mass s ## STR1 ##, ## STR1 ##, ## STR1 ##, ## STR1 ## The following is a summary of the results of this study: Einstein et al. J Oin Microbiol 50 (3): 927-937.
    权利要求:
    Claims (4)
    [0001]
    CLAIMS 1 - Method of prLD r ,, ition the area of charactersaUon of raque for r4., Minus a characterisailon r: Yune a population of at least one micro-organism e presence of at least one antimicrobier agent, this Characterization involves a mass spectrometric analysis in which the population of at least one microorganism deposited in the analysis zone is uncomfortable with at least one observation by bombardment of the analysis zone by a method of analysis. According to the technique of 7777th spectrometry by matrix-desorption-iondci spectrometry, it is characterized that the preparation of the analysis zone comprises the following successive steps: a conjugate is shown to be a cranelyseis for a crypt. : n by the technsi: c MAL. comprising: LiInc: a tryptic zone of an antimicrobial agent denoting the population of at least one microorganism on said assay enzyme in contact with an antimicrobial step incubating, To check for coridito s and for a sufficient time, tive antir ien and rinro-o LE, no analysis, in letting rinten7ttion a tape and drop on the analysis area, a matrix adapted to tachnicue IALDI.
    [0002]
    2- Process of preparation according to claim 1, characterized in that a population of a single friciii ° -orcanism to be characterized is deposited.
    [0003]
    3 - A method of preparation according to claim 1 or 2, Carac-erké in that the population of micro-orgaC5Fre dare is prepared without step this prior contact with a antiruicrobial ailment.
    [0004]
    4 - Preparation process according to any one of claims 1 to 3, characterized in that the poppy Jon microorganism (s) is obtained after a concentration step, enrichment and / or pLdcation and / or corresponds to a c 'or at a rate: the colon obtained after growth on a suitable medium, like a medium, elosed. Preparation process according to any one of Claims 1 to 4, characterized in that the antimicrobial agent is selected from a manganese to allow the identification of the staleness due to the production of lactamase, and in particular cart ase. A method of any one of claims 1 to 5, characterized in that the antimicrobial agent is antibiotic, preferably naphthalenes, cephalinic, isobrolic, and in particular among ticarcotics. -plot ::: One, the chnn, the man, the cetin, that he (cf, yime, the celtazlnurcoftalaxon, - recnoo, theopen the imioe t.oratlen according to Moon rt :. In the case of reverberations 1-6, ae, in the above, it is understood that it is isotropic, that is, to obtain the plurality of H: 2 dions. .),) u to ri3U satior by techrqnr. "IAJ DI, .. nr., M, S ti ne Z 0 Mt. Lj-lysis of an anteriority of the for-Ki-r na, realized by means of the solution r. of a method of characterizing a population 71.:7-1.1,1 minus a microorganism, the characterization comprising at least one determination of the pre-condition, if any, of a micro-elm resistant to at least one antimicrobacteristic agent characterized in that the sCDJ comprises the steps SUCCESSIVE to the preparation of the ML: A method of characterization of a S analysis (prec (i5.d.promotion, one, from claims 1 to 7), minus one year, ie mass analysis by matrix-assisted ionization and measurement of flight time In this case, a population of a microorganism deposited on the characterization zone makes it possible to conclude whether a population of an antimicrobial resistant is present in the zone of characterization. 8, because the mass spectrometric analysis according to the MALDI-TOF technique can be used to conclude whether a microbial microorganism is present. in the area of this area, in the area of the sea. This process is carried out in a vacuum-free manner, with the presence of a flame retardant in a product of the invention, nevendicab nu 9, caraere t C con tudes, in addition to , u: fan preference, the species of a popu; r to zone cl ,. tbn according to one conch u 'nun, fenar.err' of e on L ',:, Drie. r3rFi, rt puts a microphone on one: First of all on a reviirre corresponding to the ionization in ion of the analysis, a priority on the calibration, and on the caroling of a population. (I have a microgram: Artirrucrohian i-eitarty implements aru3tyse by mass spectrometry according to the technique PIA.L.1 .-, ': --- T3F corr ,,,, nnnedant to a second of the 25 The invention relates to a method for use in the field of ceramics, a method of manufacture, or a method of claim 7, wherein the method according to claim 10 or 11, in which the emptying of the family is effected. of the genus, or, nence, of the species of a population of a micro-organism, and the error of resistance; Ninfirricrobial is the most important microorganism. 13 - A method of characterizing at least one embodiment, the method comprising at least one of: a family, a genus, or, preferably, a species of a population; A micro-orgar: Nml, cleanser performs a first analysis .par-, i .-, ectrorre_r mass sekin the MALDI technique corresponds to a first ionisation of a zone of characterization corimurenant pornography-ijcro-orqa The antimicrobial effect will be determined by the technique of detecting a population with a resistant microorganism less than one antiretractor. a second analysis, in which the microbial art is reproduced below. ionization (tune minus one microdrive), the technician, the APL satin-ri corniorenii and the site of the mass control, and the second artillery FP- 15 e 5th peoure, rus r.stecd usupe tnatisatbn nistincteu of a carat n risabon comprising: p 0 pi ration of at least one rr: ro-.n -. 31'e and antimutronien, 20 'i u1, - i.e., -,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,. , TTr '' '' according to the revendicati 'r3 "Lrisé in that the popputL.:7)rr- .e comprises a sii tii, micro-l' 'nr -rrr-rrie to characterize. .1,7 - A method of c.: It is carried out as follows: (aU,.) N 13 or 14, c. F: erized in that it r a process of atio of a zone C t:! <L. d 77 the erection of an annealing plate according to ... nr (1), ... ndictatits, ins 1 to 7. 1 A "heé of functionalisatinn of a zone of analysis d ' a plate 30 acarit! n to the technique M, characterized in that the g_c ... _ _ is the z One Eyre includes: - a step of functionalities of the zone it zone carrying a dideni'antlrn microblen. 17 - Functioning method according to claim 16 characterized. in that the forming step is not a solute of the agent. anamlcroni ,,, '.. 1 iVi a seci. & ge. - Anal plate _ desi: 1 ie.:Have to obtain at least one T-JrC1r t [1e to characterize on dait ..., zone ..., 7-T spectrometry of TL. . according to tedisini u 'LD. character: that of the intimicrobian deposits an area of a region calling for a channel called tonotlonnail C 1, y i9 ti: i I zone,: antimicrobi;.: 11,1_11vi of a sec. as claimed in claim 18, characterized in that it is obtained by depositing a solution according to claim 1.8, wherein the compound is formed by the formula: It is possible to use a conductive material in the case of a reolysis (stainless steel plate 21 - Ana plate, according to 12eqis.lL claims csvscHrîsée in that lacomporte s An analysis of the fanctionnalist: a patient with an antipyretic effect.-Anal plate: according to i, sr, it is known from reveridation 18 to 21, in that the compodc mask And a second zone of functionalized grafting, which is an annular carrier, and a second zone of functionalized anatomy, which is a zone containing a second antimicrobial agent distinct from the first antimidosibien agent. a -ion due from ravdndications 18 to 22, characterized in this comprises: lu minus a sr H nn reference, intended to receive later a population of a microbial reference organism, and in that the surface of the reference analysis zone is devoid of antimicrobial agent. Use of a method according to any one of claims 18 to 2 OB717 (e) caracti according to one of the following clauses 8 to 15.5
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    同族专利:
    公开号 | 公开日
    EP3495492A1|2019-06-12|
    EP3174994B1|2019-01-30|
    WO2016016580A1|2016-02-04|
    CN106574293B|2021-12-24|
    ES2722112T3|2019-08-07|
    ZA201700534B|2018-05-30|
    JP6685279B2|2020-04-22|
    US20210324441A1|2021-10-21|
    RU2698139C2|2019-08-22|
    CN106574293A|2017-04-19|
    JP6992112B2|2022-01-13|
    US20170211123A1|2017-07-27|
    RU2017105467A3|2019-01-17|
    SG11201700450VA|2017-02-27|
    FR3024465B1|2018-03-23|
    CN114164251A|2022-03-11|
    JP2020122794A|2020-08-13|
    EP3174994A1|2017-06-07|
    KR20170041773A|2017-04-17|
    RU2017105467A|2018-08-28|
    JP2017523420A|2017-08-17|
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    法律状态:
    2015-07-17| PLFP| Fee payment|Year of fee payment: 2 |
    2016-02-05| PLSC| Publication of the preliminary search report|Effective date: 20160205 |
    2016-07-26| PLFP| Fee payment|Year of fee payment: 3 |
    2017-07-26| PLFP| Fee payment|Year of fee payment: 4 |
    2018-07-26| PLFP| Fee payment|Year of fee payment: 5 |
    2019-07-25| PLFP| Fee payment|Year of fee payment: 6 |
    2020-07-27| PLFP| Fee payment|Year of fee payment: 7 |
    2021-07-26| PLFP| Fee payment|Year of fee payment: 8 |
    优先权:
    申请号 | 申请日 | 专利标题
    FR1457389|2014-07-30|
    FR1457389A|FR3024465B1|2014-07-30|2014-07-30|CHARACTERIZATION OF MICROORGANISMS BY MALDI-TOF|FR1457389A| FR3024465B1|2014-07-30|2014-07-30|CHARACTERIZATION OF MICROORGANISMS BY MALDI-TOF|
    RU2017105467A| RU2698139C2|2014-07-30|2015-07-29|Description of microorganisms characteristics using maldi-tof|
    KR1020177005583A| KR102374464B1|2014-07-30|2015-07-29|Characterization of microorganisms via maldi-tof|
    EP15756680.3A| EP3174994B1|2014-07-30|2015-07-29|Characterization of microorganisms via maldi-tof|
    PCT/FR2015/052105| WO2016016580A1|2014-07-30|2015-07-29|Characterization of microorganisms via maldi-tof|
    CN202111498682.XA| CN114164251A|2014-07-30|2015-07-29|Characterization of microorganisms via MALDI-TOF|
    EP19152443.8A| EP3495492A1|2014-07-30|2015-07-29|Characterisation of microorganisms by maldi-tof|
    SG11201700450VA| SG11201700450VA|2014-07-30|2015-07-29|Characterization of microorganisms via maldi-tof|
    CN201580041281.8A| CN106574293B|2014-07-30|2015-07-29|Characterization of microorganisms via MALDI-TOF|
    US15/328,973| US20170211123A1|2014-07-30|2015-07-29|Characterization of microorganisms via maldi-tof|
    JP2017504745A| JP6685279B2|2014-07-30|2015-07-29|Characterization of microorganisms via MALDI-TOF|
    ES15756680T| ES2722112T3|2014-07-30|2015-07-29|Characterization of microorganisms by MALDI-TOF|
    ZA2017/00534A| ZA201700534B|2014-07-30|2017-01-23|Characterization of microorganisms via maldi-tof|
    JP2020063520A| JP6992112B2|2014-07-30|2020-03-31|Analysis of microbial characteristics via MALDI-TOF|
    US17/359,752| US20210324441A1|2014-07-30|2021-06-28|Characterization of microorganisms via maldi-tof|
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