WO1997005486A1 - Dosage immunologique rapide pour le streptococcus mutans - Google Patents

Dosage immunologique rapide pour le streptococcus mutans Download PDF

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Publication number
WO1997005486A1
WO1997005486A1 PCT/US1996/012135 US9612135W WO9705486A1 WO 1997005486 A1 WO1997005486 A1 WO 1997005486A1 US 9612135 W US9612135 W US 9612135W WO 9705486 A1 WO9705486 A1 WO 9705486A1
Authority
WO
WIPO (PCT)
Prior art keywords
antibody
streptococcus mutans
detecting
label
oral bacterium
Prior art date
Application number
PCT/US1996/012135
Other languages
English (en)
Inventor
Stephen Alden Ralls
Llyod Grant Simonson
Original Assignee
The United States Of America, Represented By The Secretary Of The U.S. Department Of The Navy
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
Application filed by The United States Of America, Represented By The Secretary Of The U.S. Department Of The Navy filed Critical The United States Of America, Represented By The Secretary Of The U.S. Department Of The Navy
Priority to EP96924689A priority Critical patent/EP0871890A4/fr
Priority to AU65078/96A priority patent/AU6507896A/en
Publication of WO1997005486A1 publication Critical patent/WO1997005486A1/fr

Links

Classifications

    • 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
    • G01N33/56944—Streptococcus
    • 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
    • G01N33/56955—Bacteria involved in periodontal diseases

Definitions

  • This invention relates to a rapid assay method and means of detecting dental caries.
  • the invention relates to a rapid assay test for detecting serovars of
  • Streptococcus mutans Streptococcus mutans.
  • Streptococcus mutans The occurrence of Streptococcus mutans is highly correlated with dental caries and this has
  • an object of this invention is an immunodiagnostic assay for rapidly
  • Streptococcus mutans especially serovars c, e, and f
  • Another object of this invention is a diagnostic method for rapidly assessing dental
  • a further object of this invention is a diagnostic method for rapidly determining whether a radiographic carious lesion is in an active disease state (positive for a certain level
  • An additional object of this invention is a device for conducting the rapid immuno
  • the device for conducting these tests is a frame or support holding non-interactive material
  • Fig. la is an embodiment of the device for the method before developing the test.
  • Fig. lb is an embodiment of the device for the method showing the test fully
  • the invention is directed to immunodiagnostic assays to identify cariogenic oral
  • antibodies provide a means of detecting antibodies for rapid clinical, chairside detection of oral
  • Streptococcus mutans in oral clinical specimens is fiilly developed and readable in under an
  • rapid test is a test that can be developed in under an hour
  • test has potential for both whole-mouth screening
  • treatment resources can be directed in the most effective and efficient manner.
  • the method is sensitive, specific and semi-
  • the method can be used in a dental operatory with positive results obtained
  • a dental caries screening device or to evaluate site-specific lesions.
  • the antibodies are prepared in rabbits
  • Antibodies are naturally produced biomolecules which
  • Antibodies are created by
  • 4,458,014 can be used to prepare the monoclonal or polyclonal antibodies. This immunization
  • these antibodies are used as reagents to aid in the rapid detection of the bacteria
  • the invention is a clinical diagnostic method using polyclonal antibodies
  • the method comprises gathering a sample suspected of
  • the sample can be gathered
  • a flow through filter type device such as marketed by or a
  • a non-inactive substrate is immobilized on the substrate and the other materials are
  • the substrate can be any of the commonly used substrates such as
  • nitrocellulose filter media nitrocellulose filter media, latex beads or any of the materials described by Oprandy.
  • the unkown sample is immobilized on the substrate, the sample is contacted with a previously
  • the antibody is prepared
  • the antibody is contacted with a
  • detectable label can be used such as enzymes (e.g., alkaline phosphatase; peroxidase;
  • Prefered labels are labels such as colloidal gold coupled to protein-A, protein-G, or some other
  • detectable labels are such as fluorescent markers, radionuclides and others.
  • the presence of a target bacterium can be identified by detecting the label
  • Streptococcus mutans instead of polyclonal rabbit serum.
  • colloidal gold or other labels such as enzymes or fluorochromes could be attached to several probes such as
  • protein A protein A, protein G, goat anti-rabbit IgG, goat-anti-mouse IgG, and others.
  • the test involves five simple and rapid steps:
  • the antigen (40 ⁇ g wet weight of Streptococcus mutans bacterial cells in 4 ⁇ l) is
  • bovine serum albumin (O.D. 1.5 at 522 nm) is added for color development. A pink
  • the New Zealand White rabbits are male, one year of age, and weigh approximately nine to ten pounds each. Three rabbits were used and were sufficient to provide the quantity
  • Antibodies to specific Streptococcus mutans serovars a-g (c, e, and f used in the present
  • Streptococcus mutans strains ATCC 25175/NCTC 10449; LM-7; and OMZ-175) microbial
  • MPL Monophosphoryl Lipid A
  • TDM Trehalose Dimycolate
  • Semm was centrifuged at 10,000 x g and the supernatant was preserved by the addition of
  • the antisemm was affimty purified by using a pleated capsule recombinant protein A affinity
  • antisera is the centrifuged at 16,000 x g for 60 min. The procedure was repeated using the
  • sobrinus isolates 6715-13 and SL-1 This reagent is the specific anti-Streptococcus mutans
  • serovars c, e, and f reagent used in the present prototype test system. This forms the antibodies that are stored for later use in the rapid assay.
  • the antibody and antigen are identical to the known antibody.
  • the known antibody is N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-N-(2-aminoethyl)-2-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N
  • test strip immobilized on a substrate, prefereably nitrocellulose media to form a test strip.
  • a substrate prefereably nitrocellulose media to form a test strip.
  • the method can be used in a dental operatory with positive

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Hematology (AREA)
  • Chemical & Material Sciences (AREA)
  • Urology & Nephrology (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Physics & Mathematics (AREA)
  • Biotechnology (AREA)
  • Virology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Cell Biology (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

Analyse permettant de repérer une bactérie présente dans la bouche, de préférence Streptococcus mutans, idéalement des sérovars c, e et f. On recueille un échantillon dont on soupçonne qu'il contient une bactérie de la bouche recherchée (cible), on immobilise toute bactérie de la bouche présente sur un substrat non inactif, on met cet échantillon en présence d'un anticorps, soit un anticorps polyclonal (antisérums animaux absorbés), soit un anticorps monoclonal, spécifique à la bactérie de la bouche recherchée (cible); on met cet anticorps en présence d'un marqueur pouvant être détecté, ce qui permet de mettre en évidence la présence de l'anticorps, de repérer le marqueur et de déterminer ainsi la présence de Streptococcus mutans dans le milieu. Le dispositif permettant d'effectuer ces analyses est un cadre ou un support contenant un matériau non interactif capable de lier les antigènes recherchés (cibles) tout en permettant l'évacuation des fluides.
PCT/US1996/012135 1995-07-28 1996-07-23 Dosage immunologique rapide pour le streptococcus mutans WO1997005486A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
EP96924689A EP0871890A4 (fr) 1995-07-28 1996-07-23 Dosage immunologique rapide pour le streptococcus mutans
AU65078/96A AU6507896A (en) 1995-07-28 1996-07-23 Rapid immunoassay for streptococcus mutans

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US50865395A 1995-07-28 1995-07-28
US08/508,653 1995-07-28

Publications (1)

Publication Number Publication Date
WO1997005486A1 true WO1997005486A1 (fr) 1997-02-13

Family

ID=24023537

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1996/012135 WO1997005486A1 (fr) 1995-07-28 1996-07-23 Dosage immunologique rapide pour le streptococcus mutans

Country Status (3)

Country Link
EP (1) EP0871890A4 (fr)
AU (1) AU6507896A (fr)
WO (1) WO1997005486A1 (fr)

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0258963A2 (fr) * 1986-06-09 1988-03-09 Ortho Diagnostic Systems Inc. Essai immunologique utilisant de l'or colloidal
US4853335A (en) * 1987-09-28 1989-08-01 Olsen Duane A Colloidal gold particle concentration immunoassay
EP0496345A1 (fr) * 1991-01-22 1992-07-29 NAGASE & COMPANY, LTD. Méthode de détection et quantification de bactéries cariogénique

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4399229A (en) * 1980-04-14 1983-08-16 Immutron, Inc. Rapid radioimmunoassay product and method of making and using same
JPH01250067A (ja) * 1988-03-30 1989-10-05 Lion Corp ストレプトコッカス・・ミユータンスの検出方法

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0258963A2 (fr) * 1986-06-09 1988-03-09 Ortho Diagnostic Systems Inc. Essai immunologique utilisant de l'or colloidal
US4853335A (en) * 1987-09-28 1989-08-01 Olsen Duane A Colloidal gold particle concentration immunoassay
EP0496345A1 (fr) * 1991-01-22 1992-07-29 NAGASE & COMPANY, LTD. Méthode de détection et quantification de bactéries cariogénique

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of EP0871890A4 *

Also Published As

Publication number Publication date
EP0871890A1 (fr) 1998-10-21
EP0871890A4 (fr) 2001-05-02
AU6507896A (en) 1997-02-26

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