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EP0168422A1 - Procede et substances permettant l'identification de micro-organismes produisant du lipopolysaccharide - Google Patents

Procede et substances permettant l'identification de micro-organismes produisant du lipopolysaccharide

Info

Publication number
EP0168422A1
EP0168422A1 EP19850900397 EP85900397A EP0168422A1 EP 0168422 A1 EP0168422 A1 EP 0168422A1 EP 19850900397 EP19850900397 EP 19850900397 EP 85900397 A EP85900397 A EP 85900397A EP 0168422 A1 EP0168422 A1 EP 0168422A1
Authority
EP
European Patent Office
Prior art keywords
antibody
lipopolysaccharide
region
kit
endotoxic
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP19850900397
Other languages
German (de)
English (en)
Inventor
Charles A. Mclauglin
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Meru Inc
Original Assignee
Meru Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from US06/590,211 external-priority patent/US4683196A/en
Application filed by Meru Inc filed Critical Meru Inc
Publication of EP0168422A1 publication Critical patent/EP0168422A1/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/12—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from bacteria
    • C07K16/1203—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from bacteria from Gram-negative bacteria
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2400/00—Assays, e.g. immunoassays or enzyme assays, involving carbohydrates
    • G01N2400/10—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • G01N2400/50—Lipopolysaccharides; LPS

Definitions

  • This invention relates to the immunological 5 detection of microorganisms. More specifically, the invention relates to methods and materials useful for the detection of a diverse group of microorganisms in a clinical sample.
  • the LPS molecule has been the subject of intense study. See for example, estphal et al. , Bacterial Lipopolysaccharides, Methods of Carbohydrate Chemistry, Vol. 0 V, Academic Press, (1965), pgs. 83-91; and Galanos et. al.. Biochemistry of Lipids II, T.W. Goodwin (Ed) , Univ. Park Press, (1977) ' , Vol. 14, at page 239.
  • the structure of the lipopolysaccharide has been, described in studies of Gram-negative bacteria such as, 5 Escherichia ⁇ oli and Salmonella typhimurium.
  • the antigen can be visualized as possessing three component regions. Proximal to and imbedded into the outer portion of the cell membrane is the so-called lipid A component. This component has been associated with the endotoxin properties Q of the molecule and is believed to be a highly conserved sequence; that is to say, a wide variety of organisms would
  • the middle region " of the LPS molecule is another conserved region, the so-called core oligosaccharide.
  • the core oligosaccharide is bipartite in structure based upon the types of sugar moieties 5 of which it is composed.
  • the innermost region which is adjacent to the lipid A component contains the unusual sugar, 3-deoxy-manno-octulosonic acid, also known as ketodeoxyoctanoic acid (KDO) .
  • KDO ketodeoxyoctanoic acid
  • the outermost region of the core structure is comprised of a region of seven-carbon
  • antisera produced thereto will be specific for each type of inducing organism.
  • Microorganisms can thus be classified into che otypes based upon their carbohydrate composition in the 0 5 region of the LPS.
  • One method is described by Merrick in U.S.- Patent 3,891,508.
  • Such a composition analysis although somewhat useful for purposes of identification is limited by not being able to discriminate between two organisms which may possess the same kinds of sugars but in different 0 proportions or in different sequences.
  • Patents 4,185,090' and 4,057,685 relate to methods of reducing the toxicity of the molecule but retaining the antigenicity of the 0 region.
  • the specificity of the 0 region in permitting the identification, of only a single species is disclosed by Wallace et. al. in U.S. Patent 4,115,543 relating to the identification of Neisseria gonorrhoeae.
  • Other approaches attempt to find other antigenic inducers and eliminate the LPS molecule from the immunizing mixture as for
  • the antigen to which the monoclonals are directed is referred to as a lipoprotein complex.
  • the lipoprotein complex is probably closely related to the Chlamydia group specific protein as characterized by
  • the LPS molecule itself has been used as a marker reagent; being chemically attached to a ligand. it permits the detection of the LPS-ligand complex by standard LPS assay method (The Limulus amebocyte assay) .
  • the subject invention provides a means for the immunological detection of an entire class of microorganisms in clinical samples.
  • the detection is accomplished by reaction of the clinical sample with a class-specific immunological reagent.
  • This reagent is an antiserum either monoclonal or polyclonal in nature, and the detection is based upon reaction of the antiserum with an antigenic determinant which is shared among all members of the detectable class of microorganisms.
  • the presence of the re-sulting immunological reaction product e.g. the antigen-antibody complex
  • the subject invention is particularly useful for the detection of a class of microorganisms which produce endotoxin or endotoxin-like molecules.
  • the shared antigen determinant in this case, is often a component of a cell-surface lipopolysaccharide.
  • Microorganisms which display this antigen and thus are members of a detectable group comprise Chlamydia, Rickettsia, and other gram- negative microorganisms such as Neisseria, Brucella, Escherichia, Salmonella and the like.
  • an antibody to the shared determinant of the detectable class is used as the "capture" antibody in a differential screening assay system.
  • the captured cells or antigens are then subsequently reacted with more specific antisera to indicate which particular genera, species or serotype of microorganism is present.
  • a first antibody is synthesized which reacts with the "gram-negative-like" portion of the endotoxic glycolipid
  • second antibody is synthesized which. eacts with a Chlamvdial genus specific determinant.
  • the first antibody can be used alone to detect all gram- negative-like microorganism by simple fluorescent or • agglutination assays.
  • the two antibodies may be used in combination in a solid phase immunoassay to discriminate the Chlamydia genera from among the other members of the reactive class.
  • the Chla vdia genus specific antibody may be used alone to detect all members of the Chvamdia genus or as the capture ' antibody in a differentia ⁇ screening assay whereby the Chlamydia genus specific antibody is used to react with all members of the genus and a second antibody or antibodies which are reactive to Chlamydia species determinants are employed to differentiate each species in the reactive genera.
  • the subject invention provides an immunological reagent for the detection of shared antigenic determinants of at least two microorganisms comprising an antibody specifically reactive to said shared determinants, more specifically an immunological reagent for the detection of two or more endotqxin or endotoxin-like lipopolysaccharide producing microorganisms comprising a monoclonal antibody specifically reactive with shared antigenic determinants of said lipopolysaccharide.
  • One such method comprises: contacting a sample containing said microorganisms or antigens thereof with an analytically ' indicatable antibody to form an im unocomplex of said microorganism or antigen thereof and said antibody, said antibody being characterized as having affinity for an antigenic determinant shared by said microorganism.
  • a two-site immunometric assay to identify an antigen in a sample comprising forming a ternary complex of a first labelled antibody, said antigen, and a second antibody, said second antibody being bound to a solid carrier insoluble under ternary complex forming conditions wherein the presence of the antigen is detected and identified by measuring either the amount of labelled antibody bound to said solid carrier or the amount of unrea ⁇ ted labelled antibody, in which the improvement comprises employing as said second antibody a monoclonal antibody characterized in having affinity for an antigenic.
  • This invention provides reagents and methods for the detection and subsequent differential identification of an entire class of microbes.
  • class is not used in its strict biological/taxonomic meaning but rather in a more general sense referring to a group of microorganisms that express a common antigenic determinant.
  • antigenic determinant, antigenic determinant site (ADS) or epitope refer to a limited, specific part of an antigen which is the inducer of antibody formation and which is the region of the antigen to which the induced antibody reacts.
  • the number of antigenic determinant sites possessed by a given antigen will vary and may range from 1 to 10 or 20 or more depending upon the size and immunological complexity of the antigen molecule.
  • antigenic determinants which are unique to a given antigen are known as specific determinants and are useful in discriminating between two different types of antigen molecules.
  • Those determinants which are present on two or more antigens are known as shared determinants and, as mentioned above, form the basis of an immunological class of antigens.
  • LPS lipopoly- saccharide
  • LPS/glycolipid molecule could be considered to be comprised of one or more separate ADSs.
  • the somatic region is considered to be a specific determinant since, because of its high variability, antibodies raised to the somatic region of one LPS would not likely cross-react with another 0 determinant; thus organisms expressing a particular LPS may be distinguished from others by virtue of the somatic ("body") types.
  • Lipid A or core region different organisms that possess endotoxin or endotoxin-like entities will have antigenetically similar regions. Thus, antibodies raised to these determinants will react with an entire class -(group) of microorganisms, those of which can express LPS regardless of somatic type.
  • somatic cell hybridization procedures may be employed to construct hybridoma cell lines capable of generating monoclonal antibody to the determinants in question. These procedures are more fully described in the
  • Gram-negative bacteria expressing LPS such as Salmonella and
  • Neisseria but also all Chlamydia species as well.
  • Gram-reactive bacteria such as Salmonella, Brucella,
  • Escherichia Neisseria, Serratia, Pasteurella, Proteus, Shigella, Klebsiella, as well as Chlamydia, and Rickettsia.
  • Antibodies of the group react specifically with members of all species within a given genera but not with organisms outside that genera. These antibodies are said to be genera-specific. -iO- Thus, by using an antibody of broad specificity, it is possible to perform a single test for a large number of LPS producing organisms merely by reacting said antibody with a clinical sample and detecting any antibody-antigen reaction which occurred by measuring the antigen and antibody- complex directly, conventionally by monitoring an analytically indicatable detection system.
  • antibodies of the subject invention can be used directly and can be modified as necessary to render them useful in any of the well-known immunological detection systems.
  • suitable methods for detection of the antigen-antibody reaction include: precipitation, agglutination, double antibody techniques, fluoroscent-, enzyme-, ferritin-, or radioactively-labelled antibodies 125I-/or similarly radioactively labelled antibodies, protein A from
  • the subject invention provides specific reagents for the reaction with the LPS/glycolipid antigen and not any particular method of indicating the resulting reaction product. The most appropriate method of indication can be determined by one skilled in the art. A wide variety of indicator systems are contemplated, and the subject invention should not necessarily be limited by a particular method of indicating the positive reaction.
  • the immunological reagent (the anti-Lipid or anti-core LPS reactive antibody of or monoclonal origin) is employed as a capture antibody in a "sandwich-type" assay as described in U.S. Patent 4,376,110. According to this embodiment, the anti-Lipid A or
  • OMFI core-reactive antibody is attached to a solid support by widely known cross-linking methods such as described by S.
  • a clinical sample which may be defined as body fluids or secretions such as blood, serum, saliva, stool, urine, milk, topical washing of skin or genitals, tissue samples or homogenates thereof and samples of culture fluids of infected cells or homogenates thereof, is reacted with the supported immunological reagent under conditions which promote the formation of an immune complex between said, supported reagent and any material containing the Lipid A or core determinants, thereby binding to the support one or more members of a variety of taxonomic groups of micro-organisms.
  • the specific identity of the constituent members of the bound population may be determined, after washing the initial reaction product to remove unbound contaminants, by subsequently reacting the bound population with specific antisera for each specific taxonomic type of interest e.g. genera-specific, species-specific or serotype-specific antisera.
  • the above described "sandwich” assay can be carried out by providing a plurality of separate reactions of the supported "capture” reagent followed by detection of the reacted sample by adding a particular genera-, species- or serotype-specific reagent to 5/02685
  • Chlamydia as well as Neisseria both organisms would be retrieved by the "capture" antibody attached to the solid support.
  • addition of two additional types of detectably labelled antibodies, one specific for Chlamydia and the other specific for Neisseria would permit the simultaneous detection of both organisms. Detection is facilitated by employing analytically distinct reagents is labelling the specific antibody.
  • the Chlamydia specific antibody may be labelled with B-gala ⁇ tosedase and the product of the ELISA reaction measure of one spectrophotometric wavelength whereas the Neisseria specific antibody labelled with alkaline phosphatase and the ELISA reaction product measured at a second wavelength.
  • Other variations employ colorimetric blending of two separate reaction products or the use of a radiometric/photospectrometric combination are also possible.
  • the following examples further illustrate various facets of the invention but are not to be viewed as limitations of the invention per se.
  • New Zealand white rabbits weighing approximately 2 kg and 8 to 10-week old BALB/c mice were injected subcutaneously with an emulsion containing equal parts of incomplete Freund's adjuvant and water in which was suspended formalin-fixed Salmonella typhimurium Re chemotype whole cells. Lyophilized whole cells were formalin fixed by suspension in 0.2% formaldehyde and incubation overnight at 25°C. They were diluted into water used for preparation of the emulsion for injection so that each rabbit received 1.0 g and each mouse, 0.1 mg. The animals were injected subsequently with the same quantities of whole cells suspended in water and given intravenously at 2 to 6-week intervals for 3 to 6 months. The animals were anesthetized and exanquinated by cardiac puncture. Sera collected from the blood was stored frozen or filtered through 0.45 urn .filters and stored at 4°C in 0.1% sodium azide. This antisera is termed anti-Re glycopid antisera.
  • acetic .acid This lipid A was adsorbed to the acetic acid-treated whole bacteria by mixing into a homogeneous suspension the lipid A and the acid-treated bacteria and then evaporating the water under reduced pressure at 50-60°C until a thick slurry was generated.
  • Anti-lipid A glycopid antisera Animals immunized with this antigenic preparation produced a sera termed anti-lipid A glycopid antisera. In certain circumstances, some of the animals received an equal mixture of untreated whole cells and acid-treated, lipid A-coated whole cells. Sera from these mice is termed anti-Re-Lipid A glycolipid antisera.
  • test antisera or normal rabbit sera or normal mouse sera at various dilutions was added to each well (100 ul/well after removal-of the saline solution used for storage) .
  • the plates were incubated 3 to 18 hours at 25 or 37°C; the sera was decanted; the plates were washed with buffered saline containing MgCl (three times, 200 ul each time) ; alkaline phophatase-conjugated antiserum to rabbit or mouse immunoglobulin (IgG and IgM) (purchased from Tago Inc. , Burlingame, CA) was added and incubated for 6 to 18 hours at 37°C.
  • IgG and IgM mouse immunoglobulin
  • the plates were again washed as before and 100 ul of -15- alkaline phosphatase substrate p-nitro-phenol phosphate in 1% diethylamine and 1 mM MgCl was added. The plates were examined visually for yellow color development or spectrophotometrically at 405 nm following incubation at 25°C for -3 hours.
  • mice were immunized with formalin-fixed elementary bodies of C ⁇ trachomatis (strain L2/434/B (L2) grown in HeLa cells (see Caldwell et al. Infect. Irnmun. 31, 1161 (1981) for preparation of purified elementary bodies) .
  • the formalin fixation and immunization scheme was as described in Example 1 above.
  • Other animals were immunized with formalin-fixed N ⁇ _ gonorrhea (laboratory strain F62 phenotyped P ++0+) in a similar immunization protocol.
  • mice Female BALB/c mice were immunized with a mixture of untreated and acid-treated cells as described in Example 1. The mice received 4 to 6 injections of immunogen and were euthanized three days after the last injection. The spleens were removed aseptically and homogenized between glass microscope slides. The cells were washed in minimum essential medium (MEM) and the red blood cells lysed by treatment for 45 seconds ' in sterile water. The cells were diluted into MEM to stop the lytic process. The spleen cells remaining were pelleted at room temperature upon centrifugation for 15 minutes at 1500 rpm in a Beckman TJ-6 table top centrifuge. To this pellet was added 2 to 5 x 10 SP2/0 myeloma cells (Institute for Medical Research,
  • the combined spleen cells and myeloma cells were centrifuged for 10 minutes at 600 rpm in the Beckman TJ-6 centrifuge at room temperature. The pellet was dislodged after decanting the MEM. The cells were then treated for 1 minute with 1.0 ml of polyethylene glycol 4000 (Sigma
  • cells from positive wells were cloned by limiting dilution.
  • the cells were diluted to 30 cells and 10 cells per ml of media; 0.1 ml of these solutions were dispensed in 96-well trays.
  • An additional 0.1 ml of media containing spleen cells (2.5 x 10 /ml) was added; the trays were wrapped in plastic film and incubated at 37°C in a humidified chamber in the presence of 5% C0_ for 2 to 6 weeks.
  • the wells were again -screened for antibody production.
  • a population was considered cloned when obtained from a well that was one of a group of wells receiving the same cell contractation only if that group had growth of hybridomas in one third or fewer wells. Generally clones were cloned 3 to 4 times to ensure single-cell origin.
  • Solid Phase Immunometric Assay The principle of detection of antigens (chlamydial endotoxin or chlamydia in this example) by a solid phase immunometric assay whereby the antigen is sandwiched between two antibodies is well documented (for review see Immunoassays for the 80s, -A. Voller, A. Bartlett and D. Bidwell, eds. University Park Press, Baltimore (1981) at page 85. Heterogeneous enzyme immunoassays by S. Avrameas) .
  • the general scheme of such an assay is 1) adsorbtion or covalent linking of the first antibody to a solid phase such as a polystyrene or nylon tube, plate or bead, 2) incubation of the test material containing the antigen which will specifically bind to the first antibody, 3) washing away unreacted antigen(s) not specifically bound, 4) addition of the second antibody which will also react with the antigen already bound to the first antibody.
  • a solid phase such as a polystyrene or nylon tube, plate or bead
  • the second antibody may be labelled by iodination with radioactive iodine, e.g., or by being conjugated with an enzyme such as alkaline phosphatase) , 5) washing an unreacted labelled second antibody from the reaction vessel, and finally 6) measuring the presence of the labelled antibody remaining in the vessel (radiometrically or enzymatically) .
  • Antibody Reactivity in Enzyme Immunoassay Protocol The various antigens were coated on the surfaces of polystyrene plates as described in Example 1. The antisera and monoclonal antibody solutions were added and processed as described. The reactivity of these antibodies to the antigens were detected using alkaline phosphatase conjugated to anti-mouse or anti-rabbit immunoglobulins.
  • N.g. Neis ' erria gonorrhoeae whole cells
  • both polyclonal and monoclonal antisera can be generated that will react with endotoxic glycolipids from a variety of sources.
  • Both anti-Re glycolipid antisera and anti-Lipid A antisera reacts with periodate-sensitive determinates in C. trachomatis elementary bodies. Weak reactivity of these antisera is seen to N _ gonorrhoeae. The periodate-sensitivity of certain of these reactions is evidence that the determinate recognized by these antisera is the endotoxic glycolipid present in the celi walls of the microbes.
  • the antisera are defined by their reactivity to purified endotoxic glycolipid and Lipid A from Salmonella - whole cells.
  • these antisera contain antibodies to to other non-endotoxic molecules in the whole cells such as proteins. Only the anti-endotoxic glycolipid antibodies bind to the chlamydial and gonococcal microbes and this binding is in certain cases prevented by pretreating the microbes with periodate which destroys the unprotected sugars of the endotoxic glycolipids within the whole cells or elementary bodies. However, if the epitope comprises a chemically modified sugar (e. g. amino sugars) the periodate treatment is ineffective. Monoclonal antibodies with reactivity similar t the polyclonal antisera can also be generated.
  • a chemically modified sugar e. g. amino sugars

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Abstract

Procédé de détection immunologique de la totalité d'une classe de micro-organismes dans des échantillons cliniques. La détection est obtenue en faisant réagir l'échantillon clinique avec un réactif immunologique spécifique d'une classe. Ce réactif est un antisérum de nature soit monoclonale soit polyclonale, et la détection se base sur la réaction de l'antisérum avec un déterminant antigénique qui est partagé par tous les membres de la classe détectable de micro-organismes. La présence du produit de réaction immunologique résultant (par exemple le complexe antigène-anticorps) peut être détectée par des systèmes de détection immunologique bien connus.
EP19850900397 1983-12-12 1984-12-11 Procede et substances permettant l'identification de micro-organismes produisant du lipopolysaccharide Withdrawn EP0168422A1 (fr)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US56012383A 1983-12-12 1983-12-12
US560123 1983-12-12
US590211 1984-03-16
US06/590,211 US4683196A (en) 1983-12-12 1984-03-16 Method and materials for the identification of lipopolysaccharide producing microorganisms

Publications (1)

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EP0168422A1 true EP0168422A1 (fr) 1986-01-22

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EP19850900397 Withdrawn EP0168422A1 (fr) 1983-12-12 1984-12-11 Procede et substances permettant l'identification de micro-organismes produisant du lipopolysaccharide

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EP (1) EP0168422A1 (fr)
WO (1) WO1985002685A1 (fr)

Families Citing this family (13)

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Publication number Priority date Publication date Assignee Title
FI853395L (fi) * 1984-09-05 1986-03-06 Cetus Corp Monoklonala antikroppar som blockerar gram-negativ bakterieendotoxin.
US4918163A (en) * 1985-09-27 1990-04-17 Pfizer Inc. Monoclonal antibodies specific for lipid-A determinants of gram negative bacteria
NZ218499A (en) * 1985-12-10 1990-04-26 Genetic Systems Corp Monoclonal antibodies against pseudomonas aeruginosa, pharmaceutical compositions and detection methods
US5281518A (en) * 1986-05-01 1994-01-25 Washington Research Foundation Detection of a unique chlamydia strain associated with acute respiratory disease
EP0577144A1 (fr) * 1986-05-01 1994-01-05 Washington Research Foundation Détection d'une souche unique de Chlamydia associée à des maladies respiratoires aigues
US4906567A (en) * 1987-01-21 1990-03-06 E. I. Dupont De Nemours And Company Non-immunochemical binding of lipopolysaccharides and sandwich assays therefor
ATE76974T1 (de) * 1987-07-14 1992-06-15 Univ Manchester Verfahren zur diagnose von infektionen, bei welchem lipopolyscaccharidantigene bestimmt werden.
US6461825B1 (en) * 1987-09-30 2002-10-08 Sanofi (Societe Anonyme) Immunometric assay kit and method applicable to whole cells
DE3843784A1 (de) * 1988-12-24 1990-06-28 Battelle Institut E V Verfahren zum nachweis von gramnegativen mikroorganismen sowie hierfuer geeignete dna-proben und antikoerper
JPH0363571A (ja) * 1989-08-02 1991-03-19 Chisso Corp 大腸菌群の検査キット
ES2067387B1 (es) * 1993-02-18 1995-11-01 Inia Procedimiento para la caracterizacion de antigenos mediante el uso cuantitativo de anticuerpos monoclonales.
US6790661B1 (en) 1999-07-16 2004-09-14 Verax Biomedical, Inc. System for detecting bacteria in blood, blood products, and fluids of tissues
WO2013049596A2 (fr) * 2011-09-30 2013-04-04 The Penn State Research Foundation Immunoessais rapides, spécifiques et sensibles pour détecter des antigènes de bactéries gram-négatif hautement variables

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US4115543A (en) * 1976-12-20 1978-09-19 Canadian Patents & Development Limited Identification of Neisseria gonorrhoeae
US4376110A (en) * 1980-08-04 1983-03-08 Hybritech, Incorporated Immunometric assays using monoclonal antibodies

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO8502685A1 *

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