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WO1997034149A1 - Methode diagnostique de mycobacteriose et trousse d'essai immunologique - Google Patents

Methode diagnostique de mycobacteriose et trousse d'essai immunologique Download PDF

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Publication number
WO1997034149A1
WO1997034149A1 PCT/EP1997/001037 EP9701037W WO9734149A1 WO 1997034149 A1 WO1997034149 A1 WO 1997034149A1 EP 9701037 W EP9701037 W EP 9701037W WO 9734149 A1 WO9734149 A1 WO 9734149A1
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Prior art keywords
lam
mycobacterial
patient
urine
diagnosing
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Ceased
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PCT/EP1997/001037
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English (en)
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Stefan Svenson
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Priority to AU20942/97A priority Critical patent/AU2094297A/en
Publication of WO1997034149A1 publication Critical patent/WO1997034149A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • 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
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56911—Bacteria
    • G01N33/5695—Mycobacteria
    • 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

  • the present invention relates to a method of diagnosing a mycobacterial disease, a method of monitoring the effects of therapeutic treatment of a mycobacterial disease and an immunoassay kit for diagnosing a mycobacterial disease in a patient.
  • mycobacteria causing diseases in man and the most important ones, which are pathogenic to humans, belong to the group Mycobacterium tuberculosis (TBC) and Mycobacterium avium complex (MAC). These bacteria have different, but closely related carbohydrate antigens on their cell walls, namely lipoarabinomannans (LAM), and arabinomannans (AM).
  • LAM lipoarabinomannans
  • AM arabinomannans
  • One method of detecting a mycobacterial infection is based on detection of antibodies against LAM in a blood or serum sample form a patient ( Theuer CP, Chaisson RE, Bias D (1989), Am. Rev. Respir. Dis. 139 (4, Part 2) : A 395).
  • antibodies against a bacterial antigen are used for diagnosing it is possible that a past infection and/or vaccination rather than an on-going infection or disease is detected.
  • Mycobacterial diseases can be therapeutically treated by administration of certain antibiotics, such as Rifampicin, Etambutol, and Isoniazid.
  • lipoarabinomannans and arabinomannans (AM), derived from Mycobacterium tuberculosis (TBC) and Mycobacterium avium complex (MAC)
  • LAM lipoarabinomannans
  • AM arabinomannans
  • TBC Mycobacterium tuberculosis
  • MAC Mycobacterium avium complex
  • the present invention is directed to a method of diagnosing a mycobacterial disease in a patient typically caused by Mycobacterium tuberculosis (TBC) or Mycobacterium avium complex (MAC), wherein, in a sample of feces, sputum or urine from said patient, the presence of a mycobacterial antigen selected from the group consisting of lipoarabino- mannans (LAM), arabinomannans (AM), and fragments of LAM and AM, is determined.
  • TBC Mycobacterium tuberculosis
  • MAC Mycobacterium avium complex
  • Said determination is preferably performed by the use of polyclonal or monoclonal antibodies directed against said mycobacterial antigen, and of an assay detecting antigen/antibody complexes formed.
  • ELISA enzyme-linked immunosorbent assay
  • RIA radioimmunoassay
  • immunoblotting immunoblotting.
  • the sample of feces, sputum or urine is pretreated by heat sterilization.
  • the invention is further directed to an immunoassay kit, which comprises optionally labeled polyclonal or monoclonal antibodies directed against a mycobacterial antigen selected from the group consisting of lipoarabino- mannans (LAM), arabinomannans (AM), and fragments of LAM and AM.
  • LAM lipoarabino- mannans
  • AM arabinomannans
  • the labels of the antibodies are selected in agreement with the diagnostic method to be used, e.g. an enzyme label when ELISA is to be used etc.
  • said antibodies directed against said mycobacterial antigen has been coupled to a carrier.
  • the carrier may be a solid support such as a plastic or glass surface, a membrane of e.g. derivatized carboxy cellulose, filter of e.g. Nylon or the like.
  • the immunoassay kit may additionally comprise an optionally labeled second monoclonal or polyclonal antibody, a positive control, a negative control, and optionally buffer solution(s) and/or washing solution(s).
  • the present invention is further directed to a method of monitoring the effects of therapeutic treatment of a mycobacterial disease in a patient typically caused by Mycobacterium tuberculosis (TBC) or Mycobacterium avium complex (MAC), wherein, in a sample of feces, sputum or urine from said patient, the presence and amount of a mycobacterial antigen selected from the group consisting of lipoarabinomannans (LAM), arabinomannans (AM), and fragments of LAM and AM, is determined at appropriate stages of said therapeutic treatment.
  • LAM lipoarabinomannans
  • AM arabinomannans
  • fragments of LAM and AM fragments of LAM and AM
  • a sandwich ELISA is used for the detection of mycobacterial antigens in a sample of urine from a tuberculosis patient.
  • the wells of a microtiter plate were coated with 100 ⁇ l of (mono or polyclonal) antibodies against lipoarabinomannan (LAM) per well. After overnight incubation at room temperature, the unbound antibody was removed and the remaining free binding sites of the wells were blocked by 200 ml 0.5% casein for 1 h at 37°C. Excess casein was removed, the plate was washed 3 times with washing buffer (0.05% Tween 20 in PBS) and 100 ⁇ l of urine samples were added.
  • washing buffer 0.05% Tween 20 in PBS
  • biotinylated anti-LAM IgG (5 ⁇ g/ml in PBS), incubate the membrane for 30 min at room temperature.
  • mice were given phosphate buffered saline (PBS). Urine samples were collected from the mice on the next day and analyzed by both catch-up ELISA 6
  • Urine samples from 20 patients with active tuberculosis and from 3 patients with both HIV and Mycobacterium avium complex (MAC) were analyzed; all became positive in the test. 18 healthy control patients were also analyzed and the results were negative. Matched patients with other diseases (30 patients) were also included in these studies, and interestingly some of these control patients (5 patients, positive in the assay of the invention) who initially were clinically asserted to be none TB showed upon follow-up to either have TB or a more or less recent history of TB.
  • LAM lipoarabinomannan
  • Purified LAM may be used as a positive control in the immunoassay of the invention, and as starting material for the preparation of monospecific polyclonal antibodies against LAM (which will be exemplified below).
  • Dry cell wall (5 g) from Mycobacterium tuberculosis is sonicated in Na-acetate buffer, pH 4.7, 5X3 min, followed by extraction with 80% phenol for 1 h at 70°C. After centrifugation for 30 min at 3500 rpm the phenolic phase is reextracted with water, the phenolic phase is discarded, and the aqueous phase is pooled with the aqueous phase from the centrifugation. The pooled aqueous phase is dialyzed against 3 x 5 liters of water overnight. Chromatography on octyl- Sepharose ® (Pharmacia , Sweden) yields 5 mg (0.1%) of LAM.
  • Both monoclonal and polyclonal antibodies may be use in the immunoassay of the invention.
  • the preparation of polyclonal antibodies starting with the bacterial cell wall, and monospecific polyclonal antibodies starting with LAM are exemplified.
  • LAM (6.5 mg) is oxidized with 0.01 M NalO 4 for 7 min at 4°C in the dark. Excessive NalO 4 is eliminated by addition of ethylene glycol. Fragments of LAM are separated by gel chromatography. Coupling of the fragments to aminoethyl Bio-Gel P-2 ( Bio-Rad, USA) through reductive amination at room temperature, pH 8, for 5 days results in a 70% yield. Blockage of excessive amino groups was performed by acetylation with Na-acetate using a water soluble carbodiimide such as EDAC at room temperature, pH 4.5, for 17 h, followed by washing and subsequent equilibration of the column with PBS. Affinity-LAM column yields purified monospecific polyclonal anti-LAM IgG, which can be used instead of monoclonal antibodies in the immunoassay of the invention.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Biomedical Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Cell Biology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Biotechnology (AREA)
  • Food Science & Technology (AREA)
  • Virology (AREA)
  • Microbiology (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Peptides Or Proteins (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

L'invention a trait à une méthode diagnostique de mycobactériose chez un patient, causée, d'ordinaire, par le bacille tuberculeux (MAC) ou le complexe MAI. On vérifie, dans un échantillon de selles, d'expectorations ou d'urine, la présence d'un antigène mycobactérien appartenant au groupe constitué de lipoarabinomannes (LAM), d'arabinomannes (AM) ainsi que de fragments de LAM et d'AM, en faisant, par exemple, intervenir des anticorps polyclonaux ou monoclonaux dirigés contre cet antigène mycobactérien et en effectuant un dosage afin de déceler les complexes antigène-anticorps formés. Il est possible de pré-traiter par une stérilisation à chaud les échantillons de selles, d'expectorations ou d'urine. Cette invention concerne également une trousse d'essai immunologique à utiliser dans le cadre de cette méthode diagnostique ainsi que la marche à suivre pour contrôler les effets d'une thérapie d'une mycobactériose.
PCT/EP1997/001037 1996-03-12 1997-03-03 Methode diagnostique de mycobacteriose et trousse d'essai immunologique Ceased WO1997034149A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU20942/97A AU2094297A (en) 1996-03-12 1997-03-03 Method of diagnosing a mycobacterial disease and immunoassay kit

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
SE9600949A SE9600949D0 (sv) 1996-03-12 1996-03-12 Method of diagnosing a mycobacterial disease and immunoassay kit
SE9600949-3 1996-03-12

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WO1997034149A1 true WO1997034149A1 (fr) 1997-09-18

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Cited By (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001027613A3 (fr) * 1999-10-12 2001-10-18 Connex Ges Zur Optimierung Von Procede de detection ameliore de micro-organismes acido-resistants dans les selles
EP1158991A4 (fr) * 1999-03-15 2003-05-14 Malaghan Inst Of Medical Res Traitement de l'asthme
EP1329718A3 (fr) * 2002-01-10 2004-06-02 Becton, Dickinson and Company Methodes et appareils pour rassembler et preparer des echantillons afin de detecter des mycobacteries et leurs antigenes
EP0952849A4 (fr) * 1996-12-31 2004-09-08 Univ New York Depistage precoce des maladies mycobacteriennes
WO2006012413A1 (fr) 2004-07-20 2006-02-02 Chemogen, Inc. Anticorps enrichi pour detecter une infection mycobacterienne, procedes pour l'utiliser et test diagnostique utilisant celui-ci
EP1710584A4 (fr) * 2003-12-22 2007-11-14 Seikagaku Kogyo Co Ltd Procede permettant de mesurer le lipoarabinomannane et applications de ce procede
EP1882939A1 (fr) * 2006-07-27 2008-01-30 The Jordanian Pharmaceutical Manufacturing Co. Ventouse de détection antigène immuno-chromatographique urinaire
WO2009117462A1 (fr) * 2008-03-18 2009-09-24 Wisconsin Alumini Research Foundation Test de criblage de culture mycobactérienne pour une bactérie complexe de mycobacterium avium
US7807182B2 (en) 1996-12-31 2010-10-05 Colorado State University Research Foundation Early detection of mycobacterial disease using peptides
US8158371B2 (en) 2006-09-08 2012-04-17 Wisconsin Alumni Research Foundation Assay for antibodies to Mycobacterium paratuberculosis
JP2014232117A (ja) * 2012-04-05 2014-12-11 株式会社ビーエル 結核菌群の免疫学的検出方法及び検出用キット
WO2016012449A1 (fr) * 2014-07-22 2016-01-28 Tbdiadirect Ab Anticorps monoclonal, procédé, kit et utilisation
US9315566B2 (en) 2011-01-24 2016-04-19 National University Of Singapore Pathogenic mycobacteria-derived mannose-capped lipoarabinomannan antigen binding proteins
WO2016130638A1 (fr) * 2015-02-10 2016-08-18 University Of Utah Research Foundation Procédés de détection d'analytes et de diagnostic de la tuberculose
WO2019186486A1 (fr) 2018-03-29 2019-10-03 Foundation Of Innovative New Diagnostics Anticorps ou combinaison d'anticorps et procédé l'utilisant permettant la détection d'un antigène associé à une mycobactérie dans un échantillon d'urine d'un sujet
CN111337665A (zh) * 2020-01-16 2020-06-26 卢氏实验室公司 一种用于检测肺结核感染的免疫层析试纸条及其制备方法
WO2021127096A1 (fr) * 2019-12-17 2021-06-24 National Jewish Health Méthodes de détection de lipoarabinomannane et de diagnostic d'infection mycobactérienne non tuberculeuse

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Publication number Priority date Publication date Assignee Title
EP0273333A2 (fr) * 1987-01-02 1988-07-06 Biotest AG Procédé pour la détection immunologique de mycobactéries dans le crachat et anticorps monoclonaux pour la réalisation de ce procédé
WO1992014155A1 (fr) * 1991-02-12 1992-08-20 Dynagen, Inc. Anticorps monoclonaux diriges contre des antigenes mycobacteriens
WO1992014156A1 (fr) * 1991-02-12 1992-08-20 Dynagen, Inc. Test d'immunofluorescence servant a detecter des antigenes mycobacteriens dans des fluides biologiques
WO1992014154A1 (fr) * 1991-02-12 1992-08-20 Dynagen, Inc. Serodiagnostic d'agglutination destine a des antigenes mycobacteriens dans des echantillons biologiques

Patent Citations (4)

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Publication number Priority date Publication date Assignee Title
EP0273333A2 (fr) * 1987-01-02 1988-07-06 Biotest AG Procédé pour la détection immunologique de mycobactéries dans le crachat et anticorps monoclonaux pour la réalisation de ce procédé
WO1992014155A1 (fr) * 1991-02-12 1992-08-20 Dynagen, Inc. Anticorps monoclonaux diriges contre des antigenes mycobacteriens
WO1992014156A1 (fr) * 1991-02-12 1992-08-20 Dynagen, Inc. Test d'immunofluorescence servant a detecter des antigenes mycobacteriens dans des fluides biologiques
WO1992014154A1 (fr) * 1991-02-12 1992-08-20 Dynagen, Inc. Serodiagnostic d'agglutination destine a des antigenes mycobacteriens dans des echantillons biologiques

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S. N. CHO ET AL.: "Detection of Mycobacterium tuberculosis antigens in sputum samples from tuberculosis patients.", ABSTRACTS OF THE GENERAL MEETING OF THE AMERICAN SOCIETY FOR MICROBIOLOGY, vol. 94, no. 0, 23 May 1994 (1994-05-23) - 27 May 1994 (1994-05-27), pages 174, XP002035609 *
S.N. CHO ET AL.: "Production of monoclonal antibodies to lipoarabinomannan-B and use in the detection of mycobacterial antigens in sputum.", YONSEI MEDICAL JOURNAL, vol. 31, no. 4, 1990, pages 333 - 338, XP002035610 *

Cited By (34)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0952849A4 (fr) * 1996-12-31 2004-09-08 Univ New York Depistage precoce des maladies mycobacteriennes
US7807182B2 (en) 1996-12-31 2010-10-05 Colorado State University Research Foundation Early detection of mycobacterial disease using peptides
EP1158991A4 (fr) * 1999-03-15 2003-05-14 Malaghan Inst Of Medical Res Traitement de l'asthme
WO2001027613A3 (fr) * 1999-10-12 2001-10-18 Connex Ges Zur Optimierung Von Procede de detection ameliore de micro-organismes acido-resistants dans les selles
EP1336850A1 (fr) * 1999-10-12 2003-08-20 Connex Gesellschaft zur Optimierung von Forschung und Entwicklung Procédé de détection amélioré de micro-organismes acido-résistants dans les selles
EP1329718A3 (fr) * 2002-01-10 2004-06-02 Becton, Dickinson and Company Methodes et appareils pour rassembler et preparer des echantillons afin de detecter des mycobacteries et leurs antigenes
EP1780547A1 (fr) * 2002-01-10 2007-05-02 Becton, Dickinson and Company Methodes et appareils pour rassembler et preparer des echantillons afin de detecter des mycobacteries et leurs antigenes
EP1710584A4 (fr) * 2003-12-22 2007-11-14 Seikagaku Kogyo Co Ltd Procede permettant de mesurer le lipoarabinomannane et applications de ce procede
US8703412B2 (en) 2003-12-22 2014-04-22 Seikagaku Corporation Method of measuring lipoarabinomannan and application thereof
US7851179B2 (en) 2003-12-22 2010-12-14 Seikagaku Corporation Method of measuring lipoarabinomannan and application thereof
WO2006012413A1 (fr) 2004-07-20 2006-02-02 Chemogen, Inc. Anticorps enrichi pour detecter une infection mycobacterienne, procedes pour l'utiliser et test diagnostique utilisant celui-ci
AP2316A (en) * 2004-07-20 2011-11-04 Chemogen Inc Enriched antibody for detecting mycobacterial infection, methods of use and diagnostic test employing same.
US7335480B2 (en) 2004-07-20 2008-02-26 Chemogen, Inc. Enriched antibody for detecting mycobacterial infection, methods of use and diagnostic test employing same
US8057797B2 (en) 2004-07-20 2011-11-15 Chemogen, Inc. Method of preparing enriched antibodies for detecting mycobacterial infection
US7615222B2 (en) 2004-07-20 2009-11-10 Chemogen, Inc. Enriched antibody for detecting mycobacterial infection, methods of use and diagnostic test employing same
EA013228B1 (ru) * 2004-07-20 2010-04-30 Чемоджен, Инк. Способ получения обогащённого поликлонального антитела, высокоспецифичного к антигену поверхностного полисахарида липоарабиноманнана микобактерий, и способы его применения
EP1882939A1 (fr) * 2006-07-27 2008-01-30 The Jordanian Pharmaceutical Manufacturing Co. Ventouse de détection antigène immuno-chromatographique urinaire
WO2008011927A3 (fr) * 2006-07-27 2008-03-13 Jordanian Pharmaceutical Mfg Coupelle pour détection immunochromatographiqe d'antigènes
US8158371B2 (en) 2006-09-08 2012-04-17 Wisconsin Alumni Research Foundation Assay for antibodies to Mycobacterium paratuberculosis
US8153383B2 (en) 2008-03-18 2012-04-10 Wisconsin Alumni Research Foundation Mycobacterial culture screening test for Mycobacterium avium complex bacteria
WO2009117462A1 (fr) * 2008-03-18 2009-09-24 Wisconsin Alumini Research Foundation Test de criblage de culture mycobactérienne pour une bactérie complexe de mycobacterium avium
US9315566B2 (en) 2011-01-24 2016-04-19 National University Of Singapore Pathogenic mycobacteria-derived mannose-capped lipoarabinomannan antigen binding proteins
EP3093667A1 (fr) * 2012-04-05 2016-11-16 BL Co., Ltd Procede et kit pour la detection immunologique de complexe mycobacerium tuberculosis
EP2835646A4 (fr) * 2012-04-05 2015-12-02 Bl Co Ltd Procédé et kit pour la détection immunologique de complexe mycobacterium tuberculosis
JP2014232117A (ja) * 2012-04-05 2014-12-11 株式会社ビーエル 結核菌群の免疫学的検出方法及び検出用キット
US10823730B2 (en) 2012-04-05 2020-11-03 Tauns Co., Ltd. Method and kit for immunological detection of Mycobacterium tuberculosis
EP3499237A1 (fr) * 2012-04-05 2019-06-19 Tauns Co., Ltd. Procédé et kit de détection de immunologique du complexe de mycobacterium tuberculosis
US10883988B2 (en) 2012-04-05 2021-01-05 Tauns Co., Ltd. Method and kit for immunological detection of Mycobacterium tuberculosis complex
US10830769B2 (en) 2012-04-05 2020-11-10 Tauns Co., Ltd Method and kit for immunological detection of Mycobacterium tuberculosis
WO2016012449A1 (fr) * 2014-07-22 2016-01-28 Tbdiadirect Ab Anticorps monoclonal, procédé, kit et utilisation
WO2016130638A1 (fr) * 2015-02-10 2016-08-18 University Of Utah Research Foundation Procédés de détection d'analytes et de diagnostic de la tuberculose
WO2019186486A1 (fr) 2018-03-29 2019-10-03 Foundation Of Innovative New Diagnostics Anticorps ou combinaison d'anticorps et procédé l'utilisant permettant la détection d'un antigène associé à une mycobactérie dans un échantillon d'urine d'un sujet
WO2021127096A1 (fr) * 2019-12-17 2021-06-24 National Jewish Health Méthodes de détection de lipoarabinomannane et de diagnostic d'infection mycobactérienne non tuberculeuse
CN111337665A (zh) * 2020-01-16 2020-06-26 卢氏实验室公司 一种用于检测肺结核感染的免疫层析试纸条及其制备方法

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AU2094297A (en) 1997-10-01
SE9600949D0 (sv) 1996-03-12

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