WO1999032656A1 - Evaluation colorimetrique de la sensibilite d'helicobacter pylori a des substances antimicrobiennes - Google Patents

Evaluation colorimetrique de la sensibilite d'helicobacter pylori a des substances antimicrobiennes Download PDF

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Publication number
WO1999032656A1
WO1999032656A1 PCT/IT1998/000367 IT9800367W WO9932656A1 WO 1999032656 A1 WO1999032656 A1 WO 1999032656A1 IT 9800367 W IT9800367 W IT 9800367W WO 9932656 A1 WO9932656 A1 WO 9932656A1
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Prior art keywords
helicobacter pylori
colour
culture medium
colourimetric
sensitivity
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PCT/IT1998/000367
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English (en)
Inventor
Alessandro Zuccato
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Consortia Laboratories
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Publication date
Application filed by Consortia Laboratories filed Critical Consortia Laboratories
Priority to AU17831/99A priority Critical patent/AU1783199A/en
Publication of WO1999032656A1 publication Critical patent/WO1999032656A1/fr

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    • 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
    • C12Q1/58—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving urea or urease
    • 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
    • C12Q1/18—Testing for antimicrobial activity of a material
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/62—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving urea
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/90—Enzymes; Proenzymes
    • G01N2333/914—Hydrolases (3)
    • G01N2333/978—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5)
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/90—Enzymes; Proenzymes
    • G01N2333/914—Hydrolases (3)
    • G01N2333/978—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5)
    • G01N2333/98—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5) acting on amide bonds in linear amides (3.5.1)

Definitions

  • the present invention relates to a colourimetric method suitable for the evaluation of the sensitivity of Helicobacter pylori bacteria to single antimicrobial substances, said method being applicable to a liquid culture medium.
  • a further object of the present invention is that of proposing a
  • the present invention finds a particularly advantageous application in the field of diagnostics at the microbiology laboratory level, as it renders the Helicobacter pylori senitivity test to antibiotics extremely simple, and according to a preferred form of embodiment of the present invention, it further makes it possible to entirely carry it out
  • Helicobacter pylori is a urease positive bacterium that infects human stomach and is accountable for as the main aetiologic agent of gastroduodenal diseases.
  • antibiotics like amoxycillin, 30 clarythromycin, tetracycline; chemioterapeutics like metronidazole; and antiacids like omeprazole, lansoprazole, ranitidine.
  • a first important aspect of the above operational method is the fact that the above mentioned pharmaceuticals must be administered over a period of time generally ranging between few days and few weeks.
  • Helicobacter pylori is always and constantly sensitive to some types of antibiotics, among which for example amoxycillin. It is further known on the other hand that other antibiotics like for example metronidazole and chlarythromycin exhibit a very variable biological activity towards it, although they are meant to be molecules of choice for the eradication of the infection deriving from it.
  • the infecting strain should be isolated by culture and tested in vitro in order to assess its sensitivity to antibiotics so as to rule out the employment of ineffective pharmaceuticals and therefore be able to immediately devise the most suitable therapy whose features derive from those of the specific case under treatment.
  • antibiograms are carried out on solid substrates by means of plates whereon the bacteria to test have been implanted.
  • a further operational method is moreover represented by the E-test® that is basically a quantitative variation of the plate diffusion test outlined above.
  • US Pat. N° A-4,748,1 13 discloses a method of the colourimetric type which is suitable for detecting urease, aiming at determining the presence of Helicobacter pylori in a gastric mucose sample. It is known in fact that as the proliferation of Helicobacter pylori causes an increase in the production of the urease enzyme, it also yields to a remarkable increase of pH in the culture medium which is due to the fact that the bacterium hydrolyses urea to yield carbon dioxide and ammonium ions, which are well known for their alkalinity.
  • the increase in the alkalinity of the medium yields to a colour change and it is therefore an expression of bacterial growth.
  • a sample of gastric mucose is placed in an aqueous solution containing urea at a concentration ranging between 10 and 40 g/1, a coloured pH indicator, preferably Phenol red, and a bactericide geared towards inhibiting the growth of other urease positive bacteria.
  • the above document discloses therefore a method of the colourimetric type which is exclusively suitable for diagnosing the presence of Helicobacter pylori in the tested sample, said presence being shown by a change in colour of the colour indicator employed following an increase in urease production as this is secreted by Helicobacter pylori.
  • the sample is remarkably enriched with Helicobacter pylori, by incubation in a specific culture medium which has been suitably chosen on account of its selectivity towards Helicobacter pylori.
  • microdilution test The existence of a microdilution test is further known, said test being suitable for use on a culture sample of the liquid type and for the determination of the responsivity of Helicobacter pylori to antibiotics. Diagnostic parameter of this test is the turbidity of the sample.
  • That method consists in fact in the dilution of the sample by placing it on multiwell microplates and in the successive measurement of the turbidity of the solution which is used as an index of bacterial growth as it took place within the sample contained in said wells.
  • It is an object of the present invention therefore to propose a method for the determination of the sensitivity of Helicobacter pylori to certain pharmaceuticals, said method having the same advantageous features of the methods which are known to be applicable to a liquid medium, such as the accuracy of the results and method standardisation, also bearing enhanced features as regards ease of handling, rapidity and cost effectiveness with respect to the analyses that are traditionally carried out on a substrate of the solid type.
  • the present invention therefore relates to a colourimetric type method which was devised in order to be applied to a culture medium of the liquid type, said method being suitable for the determination of Helicobacter pylori's sensitivity to the antimicrobial substances that are meant to be tested.
  • colour change towards alkalinity in other words towards bacterial growth, as described above, is an index of resistance to antibiotics, whereas a scarce colour change, or the absence of it, is a clear evidence that an inhibition of bacterial growth is taking place, therefore that there is a sensitivity to antibiotics.
  • the present invention relates to a kit by which said method can be advantageously and/herously applied.
  • one or more Helicobacter pylori colonies are suspended in a predetermined amount of a culture medium of the liquid type, after their identification has been made in a primary culture made up starting from a sample of human gastric mucose.
  • pH colour indicator such as phenol red and urea as a substrate are added.
  • the culture substrate has acquired a yellow or yellow orange colour, since its initial pH ranges between 4.2 and 6.
  • the suspension is then scattered among the plastic material microwells whose size is the same as that generally employed for immunoenzyme tests.
  • a predetermined number of said microwells contains different types and amounts of the antimicrobial substances to be tested, whereas other microwells are not filled with any antimicrobial substance to be tested as they have to uniquely supply a reference parameter.
  • the inspection is an examination with the naked eye of the colour acquired by the microwells.
  • Helicobacter pylori in one or more wells is shown by a change of their colour, that turns to red or red-blue from yellow or yellow-orange.
  • the colour change of the indicator occurs as a consequence of the pH increase within the culture medium that coincides with a substantial and remarkable bacterial growth.
  • the microbial suspension under exam is scattered among wells containing different amounts of more than one antibiotics to be tested and it will be thus possible to determine the minimum concentration of antibiotics that is necessary to inhibit the growth of Helicobacter pylori (MIC).
  • the colour change of the culture medium is determined by a spectrophotometric reading of the wells.
  • measurements are carried out by an immunoenzymatic dosage microplate detector, and they are advantageously elaborated by a suitable electronic calculator.
  • an electronic calculator further makes it possible to carry out the automatic interpolation of the optical densities measured in the single wells containing the antibiotics at different concentrations, and to work out the MIC with a high degree of precision.
  • a suitable software it is also possible to automatise reporting and printing the results on paper.
  • the pH colour indicator used is phenol red and measurements are made using a 545 nm wavelength. It is clear that the present invention does not result to be limited to the utilisation of microwells as described above, but whatever other type of vessel is employable, as well as there results to be possible to carry out the measurements of which above, using other types of spectrophotometric equipment of the automatic type.
  • a further object of the present invention is that of proposing a suitably equipped kit so as to make it possible to carry out the method detailed above.
  • Said kit comprises: a) a plurality of vessels made of transparent material containing different antimicrobial substances in suitably predetermined amounts, said vessels being advantageously assembled in printed modules and further being packed in a sterile environment. b) A culture medium for Helicobacter pylori containing urea and a pH coloured indicator.
  • the culture medium can also be supplied ready for use or in the form of a lyophile preparation to be resuspended in sterile distilled water.
  • the present invention provides a method with a kit relative thereto, based on the urease activity of Helicobacter pylori, said parameter being measured as a growth index of said bacterium in order to test its sensitivity to antibiotics in a culture medium of the liquid type, therefore not aiming at anything diagnostic, in order to evaluate the occurrence or lack thereof of Helicobacter pylori in a given culture medium, as it was mentioned in the state of the art outlined above.
  • DESCRIPTION OF SOME FORMS OF EMBODIMENT The Helicobacter pylori isolate to test is generally obtained from a culture, for example a blood agar plate previously inoculated with gastric material excised from a patient during an endoscopic exam.
  • Helicobacter pylori in a culture is accomplished by the well known methods based on the determination of the morphological features (curved or spiralled shape), Gram negativity, biochemical features (catalase, urease and phosphatase positivity; hyppurate hydrolisis and nitrate reduction negativity).
  • the bacterium can further be identified by resorting to the immunologic method which is disclosed in Italian patent application N. VR96A000109 in the name of the
  • the Applicant further claims an antigen with an apparent molecular weight of 16 ⁇ 2 kDa of Helicobacter pylori as an indicator of the occurrence of Helicobacter pylori and an immunological type method which is suitable for the detection of said antigen.
  • the following phases can be identified: a) supplying a generic sample for analysis as a solid, liquid or in suspension; b) supplying a monoclonal and/or polyclonal antibody in a biologically active form which immunoreacts with the antibody having an apparent molecular weight of 16 ⁇ 2 kDa of Helicobacter pylori; c) mixing the sample with the antibody of step b) in order to obtain an immunoreaction mixture; d) keeping the mixture in the suitable conditions for favouring the reaction between antigen and antibody for a period of time which ranges between few minutes and some hours.; e) making sure that a reaction between antigen and antibody has alraedy taken place, and determining whether within the sample under exam there is found to occur the antigen with an apparent molecular weight of 16 ⁇ 2 kDa corresponding to
  • Helicobacter pylori and, possibly, determining the concentration thereof. Once the occurrence of Helicobacter pylori has been detected according to one of the methods detailed above, according to the present invention, one or more colonies of Helicobacter pylori are resuspended in a liquid type culture medium test tube.
  • the culture media of the liquid type that can be employed for the growth of Helicobacter pylori are well known to the skilled in the art, and anyone of them can be functionalously be resorted in order to carry out the test according to the present invention.
  • a preferred culture medium is made of Brucella broth additioned with bovine foetal serum and Isovitalex®.
  • Urea at a concentration ranging between 1 g/1 and 4 g/1, preferably between 2 and 3 g/1, is added to the liquid culture medium, said urea representing the enzyme substrate for bacterial urease.
  • the amount of urea given above is sufficient to produce an appreciable increase in pH whenever there is the occurrence of growth of Helicobacter pylori, and it is not of any obstacle to the growth of the bacterium itself.
  • the culture medium is additioned with phenol red to obtain a concentration which is higher than 0. 001% (g/100 ml), more preferably between 0.005% and 0.02%.
  • pH indicators can be used so long as in the culture medium used they change colour at a pH range between 5.5 and 9.0; among these are for example bromothymol blue, para-nitrophenol, neutral red, cyanin, cresol red, metacresol purple and thymol blue.
  • the culture medium where the test is carried out according to the invention has a pH which is preferably in the range between 4.2 and 6.0, more preferably between 5.0 and 5.5.
  • the medium has a pH that can suitably be adjusted by the addition of acids or bases.
  • 1 litre of Brucella broth culture medium can be taken from pH 7.0 (red coloured medium), to pH 5.5 (yellow coloured medium) by the addition of about 8 ml of 2N HCl.
  • the microwells used are made of polystyrene, and they are located in such a way to form strips each with 8 microwells (said strips being located in suitable support frames), and they are commonly used for traditional immunoenzyme laboratory assays. 16 well strips are also emplyable (2 X 8) or entire 96 well plates which are normally employed in cell culture immunoenzyme assays.
  • test according to the present invention can be carried out in any other type of sterile plates, or in any other vessel such as a test tube or whatever vessel made of plastic material, both rigid and flexible.
  • each of the dry and sterile microwells which are transparent looking, has a specific antibiotics present in a predetermined amount, sticking to its inside surface, said antibiotics and said amounts being different between successive microwells.
  • one or more microwells do not contain any antimicrobial substance.
  • microwells have in fact the role of providing a test positive control, that is no growth inhibition whatsoever, as described below within the present specification.
  • one or more microwells result to have been prepared so as to contain a mixture of antimicrobial substances which are consistently lethal against Helicobacter pylori, said microwells aiming at providing a test negative control, that is the greatest growth inhibition, as it will be made clearer below.
  • metronidazole For example, if 0.5 ⁇ g metronidazole is present in a microwell, sticking onto the inner surface of the well itself, when this is filled with 100 ⁇ l of culture medium, metronidazole is dissolved yielding a concentration equalling or slightly below 5 ⁇ g/ml. Tha amount of antimicrobial substances contained in the microwells can be worked out in such a way as to obtain final concentrations ranging between 0.0001 and 50 ⁇ g/ml.
  • the assay by the assay, the effect of the antimicrobial substance to be tested on Helicobacter pylori is analysed, at the concentration that the skilled in the art believe to be the end concentration that defines the degree of sensitivity and/or resistance of Helicobacter pylori to that given antimicrobial substance.
  • breakpoint concentration
  • the antimicrobial substance of choice is then assayed even at higher or lower concentrations than the "breakpoint" concentration.
  • the wells are made to undergo incubation in a microaerobic or anaerobic atmosphere.
  • This incubation can also be carried out in a specific incubator for cell cultures and with an atmosphere containing 5-10% C0 2 or it can be carried out in an ambient atmosphere if suitable liquid media are employed that make it possible for Helicobacter pylori to grow outside the incubator itself.
  • the wells are inspected in order to check the colour of the culture medium, and according to different operational embodiments of the present invention, the inspection can be carried out both at a glance and by an instrumental examination of the spectrophotometric type.
  • the evaluation is anyhow carried out by comparison of the colour of the reference microwell, that is containing a predetermined concentration of a predetermined antibiotics, with the colour of the positive and negative control wells.
  • Helicobacter pylori proliferates without finding any hindrance, in so doing causing the colour change of the medium yielded to by urease production.
  • the positive control colour is the evidence of a 100% growth, according to the methodology so far described.
  • the urease present therein is that corresponding to that contained in the initial bacterial inoculate.
  • the Helicobacter pylori strain under assay can be defined as being sensitive to that given antibiotics at that given concentration.
  • the operational method according to the present invention comprises the assessment of the Helicobacter pylori strain under consideration at different concentrations of antimicrobial substance, therefore providing minimum inhibiting concentration values for the strain under assay. It is clear that different strains of Helicobacter pylori have different MIC values, with respect to the same type of antimicrobial substance.
  • the antimicrobial concentration relative to that given well corresponds to MIC; c) all the wells have a colour that is the same as that of the negative control, therefore the strain under consideration is sensitive and MIC results to be lower than the minimum concentration of the antibiotics which is being assayed.
  • phenol red is used as a pH indicator
  • measurements are preferably carried out using a 545 nm electromagnetic radiation as at this wavelength optical density differences on reading between the positive control (red) and the negative control (yellow) are higher.
  • measurements are made by availing of a microplate detector that offers numerous advantages when compared with simple naked eye measurements.
  • a first advantage is given by the greater sensitivity attained by spectrophotometric measurements, which is a particularly important aspect when it turns out to be necessary to quantify MIC.
  • 2 wells respectively containing 4 and 8 ⁇ g of antibiotics for each ml of medium may seem to have the same colour as that of the negative control well when looked at with the naked eye, and MIC would then accountably be given a value approximately equalling 4 ⁇ g/ml.
  • MIC would be quantified as equal to or greater than 8 ⁇ g/ml.
  • EXAMPLE I Three patients were subjected to gastroscopy and a sample of gastric mucose (or stomach lining), excised from each patient was inoculated onto a blood agar plate additioned with antibiotics suitable for ensuring a selective growth of Helicobacter pylori.
  • the three plates are then incubated in microaerophiles in a jar at 37° C for three days. After that incubation period has elapsed, from an examination of the plates, there appear to be evident numerous little translucent colonies and one or more colonies are picked from each plate by means of a sterile loop.
  • the colonies are identified as Helicobacter pylori colonies and after that the test for sensitivity to antimicrobials is carried out according to the present invention.
  • the three Helicobacter pylori isolates were assayed in order to test their sensitivity to metronidazole and chlarythromycin.
  • the culture medium used was Brucella broth additioned with 2 mg/ml urea
  • the amounts of the two antimicrobials chosen were positioned in the microwells so as to yield for the two strips each with eight microwells, to the final situation shown in Table I, after the addition of a microbial suspension.
  • wells Al and A2 do not contain any antimicrobial substance and they can therefore be taken to be the positive control, that is the situation when an undisturbed growth of the bacterium takes place.
  • HI and H2 wells instead both contain a mixture of amoxycillin and gentamycin, each with a 5 ⁇ g/ml final concentration, and they can be taken as the so called negative control, that is a total inhibition of bacterial growth.
  • the two antimicrobial substances to test have been supplied in serial concentrations according to a log 2 progression, as detailed in Table I.
  • the six strips two for each of the three samples under test, were placed in a suitable frame, and they were then covered with a sterile 96 well plate lid, and incubated for a 16-48 hour period in an incubator set at 37°C and with a 5% CO : concentration.
  • the plate was then scrutinised for the first time after 16 hours.
  • Example II an operational strategy similar to what was done in Example I was followed, except for the difference that the assessment of the results was carried out with the naked eye.
  • EXAMPLE III Three patients were subjected to gastroscopy and a sample of gastric mucose (otherwise known as stomach lining) picked from each patient, was inoculated onto a blood agar plate additioned with antibiotics suitable for guaranteeing a selective growth of Helicobacter pylori.
  • the three plates were then inoculated in a microaerophile jar at 37°C for three days: from an assay of the plates, there resulted to be evident a number of small translucent colonies, and one or more colonies were picked from each plate by means of a sterile loop.
  • the identification of the colonies was conducted using the immunological method which is the object of Italian patent application n° VR96A000109 in the name of the Applicant on the basis of which the colonies were identified as Helicobacter pylori.

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Abstract

Procédé colorimétrique qu'on peut appliquer avantageusement à un milieu de culture liquide et qui permet d'évaluer in vitro la sensibilité et la résistance d'Helicobacter pylori à des substances pharmaceutiques antimicrobiennes. Ce procédé est basé sur la détection colorimétrique d'une croissance bactérienne d'Helicobacter pylori provenant d'une augmentation d'uréase bactérienne, ladite variation colorée étant rendue possible par un indicateur coloré de pH injecté dans le milieu de culture. Trousse colorimétrique servant à évaluer la sensibilité d'Helicobacter pylori et/ou sa résistance à des substances pharmaceutiques antimicrobiennes, ladite évaluation étant exécutée soit à l'oeil nu, soit par des moyens spectrophotométriques, ladite trousse comprenant: (a) une pluralité de puits microscopiques et/ou de réservoirs en matériau transparent contenant des substances antimicrobiennes prédéterminées à des concentrations prédéterminées de façon appropriée, lesdits réservoirs étant assemblés avantageusement en modules imprimés et, de plus, conditionnés de manière stérile; (b) un milieu de culture d'Helicobacter pylori contenant carbamide et un indicateur coloré de Ph.
PCT/IT1998/000367 1997-12-23 1998-12-16 Evaluation colorimetrique de la sensibilite d'helicobacter pylori a des substances antimicrobiennes WO1999032656A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU17831/99A AU1783199A (en) 1997-12-23 1998-12-16 Colorimetric assessment of the sensitivity of (helicobacter) (pylori) to antimicrobial substances

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Application Number Priority Date Filing Date Title
IT97VR000122A IT1297510B1 (it) 1997-12-23 1997-12-23 Metodo colorimetrico per valutare la sensibilita' dell'helicobacter pylori a sostanze antimicrobiche e kit per attuare tale metodo.
ITVR97A000122 1997-12-23

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Cited By (11)

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WO2000008204A2 (fr) * 1998-07-31 2000-02-17 Byk Gulden Lomberg Chemische Fabrik Gmbh Procede d'identification de substances modulant des mecanismes dependant de urei du metabolisme a helicobacter pylori
GB2378505A (en) * 2001-08-10 2003-02-12 Kanto Kagaku Detecting presence of microorganisms
WO2007090683A1 (fr) * 2006-02-08 2007-08-16 Dsm Ip Assets B.V. combinaison d'un lecteur et d'un incubateur
WO2008126111A1 (fr) * 2007-04-12 2008-10-23 Alma Mater Studiorum-Università Di Bologna Test rapide de chimiosensibilité d'helicobacter pylori
CN103792349A (zh) * 2012-11-05 2014-05-14 江苏维赛科技生物发展有限公司 硝基咪唑类药物残留快速检测试纸条
WO2015128650A1 (fr) * 2014-02-26 2015-09-03 Spectromics Limited Procédé d'analyse d'un échantillon comprenant un micro-organisme d'intérêt
CN111289505A (zh) * 2020-03-16 2020-06-16 南京康容健康科技有限公司 一种幽门螺杆菌检测试剂盒
CN112695066A (zh) * 2021-01-29 2021-04-23 郑州安图生物工程股份有限公司 一种胃幽门螺杆菌培养鉴定药敏试剂盒及检测方法
CN112877245A (zh) * 2021-02-23 2021-06-01 海南省临床微生物学检测与研究中心 幽门螺杆菌敏感、快速培养菌落形成及其效能评价方法
CN115825055A (zh) * 2023-02-16 2023-03-21 山东众之康生物科技有限公司 一种幽门螺杆菌检测试剂及检测方法
WO2023228142A1 (fr) * 2022-05-27 2023-11-30 Papyrus Diagnostics Private Limited Dispositif et procédé de détection phénotypique de résistance antimicrobienne aux médicaments utilisant la microfluidique à base de papier

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US4748113A (en) * 1985-06-13 1988-05-31 Marshall Barry J Compositions and methods for the diagnosis of gastrointestinal disorders involving urease
US5439801A (en) * 1994-02-14 1995-08-08 Chek-Med Systems, Inc. Test composition for the rapid detection of helicobacter pylori in gastric biopsy tissue
EP0689842A1 (fr) * 1994-06-27 1996-01-03 DIMOTECH Ltd. Utilisation d'un extrait organique de cannelle pour l'inhibition d'helicobacter pylori
US5498528A (en) * 1994-06-10 1996-03-12 King; Wing Detection of helicobacter pylori
DE19547708A1 (de) * 1995-12-20 1997-06-26 Behringwerke Ag Verfahren zur Reinigung von Urease aus Helicobacter pylori

Patent Citations (5)

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Publication number Priority date Publication date Assignee Title
US4748113A (en) * 1985-06-13 1988-05-31 Marshall Barry J Compositions and methods for the diagnosis of gastrointestinal disorders involving urease
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WO2000008204A2 (fr) * 1998-07-31 2000-02-17 Byk Gulden Lomberg Chemische Fabrik Gmbh Procede d'identification de substances modulant des mecanismes dependant de urei du metabolisme a helicobacter pylori
WO2000008204A3 (fr) * 1998-07-31 2000-05-11 Byk Gulden Lomberg Chem Fab Procede d'identification de substances modulant des mecanismes dependant de urei du metabolisme a helicobacter pylori
GB2378505A (en) * 2001-08-10 2003-02-12 Kanto Kagaku Detecting presence of microorganisms
GB2378505B (en) * 2001-08-10 2004-01-14 Kanto Kagaku Method for detecting microorganisms, particularly fungi, and detection kit
US6984500B2 (en) 2001-08-10 2006-01-10 Tomota Nakano Method for detecting microorganisms and detection kit
US8072602B2 (en) 2006-02-08 2011-12-06 Dms Ip Assets B.V. Combination of reader and incubator
EP2336350A1 (fr) * 2006-02-08 2011-06-22 DSM IP Assets B.V. Combinaison d'un lecteur et d'un incubateur
WO2007090683A1 (fr) * 2006-02-08 2007-08-16 Dsm Ip Assets B.V. combinaison d'un lecteur et d'un incubateur
US8325341B2 (en) 2006-02-08 2012-12-04 Dsm Ip Assets B.V. Combination of reader and incubator
WO2008126111A1 (fr) * 2007-04-12 2008-10-23 Alma Mater Studiorum-Università Di Bologna Test rapide de chimiosensibilité d'helicobacter pylori
CN103792349A (zh) * 2012-11-05 2014-05-14 江苏维赛科技生物发展有限公司 硝基咪唑类药物残留快速检测试纸条
US10570436B2 (en) 2014-02-26 2020-02-25 Spectromics Limited c/o V J Hancock & Co., Ltd. Method of analysing a sample including a microorganism of interest
WO2015128650A1 (fr) * 2014-02-26 2015-09-03 Spectromics Limited Procédé d'analyse d'un échantillon comprenant un micro-organisme d'intérêt
CN111289505A (zh) * 2020-03-16 2020-06-16 南京康容健康科技有限公司 一种幽门螺杆菌检测试剂盒
CN112695066A (zh) * 2021-01-29 2021-04-23 郑州安图生物工程股份有限公司 一种胃幽门螺杆菌培养鉴定药敏试剂盒及检测方法
CN112695066B (zh) * 2021-01-29 2023-04-25 郑州安图生物工程股份有限公司 一种胃幽门螺杆菌培养鉴定药敏试剂盒及检测方法
CN112877245A (zh) * 2021-02-23 2021-06-01 海南省临床微生物学检测与研究中心 幽门螺杆菌敏感、快速培养菌落形成及其效能评价方法
WO2023228142A1 (fr) * 2022-05-27 2023-11-30 Papyrus Diagnostics Private Limited Dispositif et procédé de détection phénotypique de résistance antimicrobienne aux médicaments utilisant la microfluidique à base de papier
CN115825055A (zh) * 2023-02-16 2023-03-21 山东众之康生物科技有限公司 一种幽门螺杆菌检测试剂及检测方法

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