WO1996032646A1 - Procede de depistage du vih-tat - Google Patents

Procede de depistage du vih-tat Download PDF

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Publication number
WO1996032646A1
WO1996032646A1 PCT/DE1996/000641 DE9600641W WO9632646A1 WO 1996032646 A1 WO1996032646 A1 WO 1996032646A1 DE 9600641 W DE9600641 W DE 9600641W WO 9632646 A1 WO9632646 A1 WO 9632646A1
Authority
WO
WIPO (PCT)
Prior art keywords
tat
hiv
body sample
detergent
antibody
Prior art date
Application number
PCT/DE1996/000641
Other languages
German (de)
English (en)
Inventor
Peter Krammer
Michael Westendorp
Rainer Frank
Christina Berndt
Original Assignee
Deutsches Krebsforschungszentrum Stiftung des öffentlichen Rechts
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 Deutsches Krebsforschungszentrum Stiftung des öffentlichen Rechts filed Critical Deutsches Krebsforschungszentrum Stiftung des öffentlichen Rechts
Priority to EP96909957A priority Critical patent/EP0820595A1/fr
Priority to JP8530644A priority patent/JPH11503524A/ja
Publication of WO1996032646A1 publication Critical patent/WO1996032646A1/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
    • G01N33/56983—Viruses
    • G01N33/56988—HIV or HTLV

Definitions

  • the present invention relates to a method for the detection of HIV-TAT in a body sample and a kit that can be used therefor.
  • T cell apoptosis AIDS T cells often suffer programmed cell death. This cell death is called T cell apoptosis. Recent work by the applicant indicates that T cell apoptosis in AIDS is enhanced by an HIV protein called TAT.
  • the present invention is therefore based on the object of providing a method with which HIV-TAT can be detected in a body sample.
  • body sample includes body samples of all types of humans and animals. In particular, these are blood, sputum, urine, stool, cerebrospinal fluid, bile, gastrointestinal secretions, organ punctures, biopsies and lymphatic fluid.
  • a body sample is then washed with a common detergent, e.g. NP-40 or Triton X-100, incubated.
  • a common detergent e.g. NP-40 or Triton X-100
  • Several detergents can also be used, simultaneously or in succession. It is beneficial to use a total of 1% detergent.
  • high salt is added to the body sample. This can be a common salt, e.g. NaCI, be.
  • salts can also be used, simultaneously or in succession. It is expedient to use a total of 0.1-1 M salt.
  • the body sample is then subjected to size fractionation.
  • Common methods e.g. Fractionation by centrifugation. Fractionation. Columns from Filtron, Northborough, MA, USA are used. It is favorable to carry out the fractionation in a range of 5-30 kD.
  • HIV-TAT includes any TAT and fragments thereof derived from viruses that can cause HIV infection and / or can cause AIDS.
  • the term also relates to any TAT and fragments thereof that have been synthesized.
  • the antibody can be polyclonal or monoclonal, a monoclonal antibody being preferred.
  • the antibody can be synthetic, parts or parts of it that are not necessary for the detection of HIV-TAT being missing in whole or in part or these parts being replaced by others which impart further favorable properties to the antibody. It can also be advantageous if different anti-HIV
  • TAT antibodies can be used simultaneously or sequentially. It is particularly expedient to incubate the anti-HIV-TAT antibody (s) in parallel with the above incubation with a defined TAT standard, whereby a quantitative detection of HIV-TAT in the body sample is facilitated.
  • Incubation of the body sample with the anti-HIV-TAT antibody (s) as well as incubation of the latter with a TAT standard can be part of a conventional method, such as a Western blot, an ELISA, e.g. Sandwich or competition ELISA, immunofluorescence or immunoprecipitation.
  • the antibody can, if it is appropriate, be labeled or can be used in combination with a labeled antibody directed against it.
  • the method according to the invention is characterized in that HIV-TAT can be detected in a body sample.
  • the process is specific and can be carried out quickly. It is therefore ideal for diagnosing HIV infection and / or for tracking AIDS.
  • the latter also has the great advantage of directly monitoring the effects of therapeutic measures, particularly against HIV-TAT.
  • a kit for the detection of HIV-TAT in a body sample is also provided.
  • a kit for the detection of HIV-TAT in a body sample advantageously includes the following:
  • the figure shows the specific detection of HIV-TAT in body samples from HIV-1 infected people.
  • the invention is illustrated by the following example.
  • Serum samples were taken from 33 people infected with HIV-1. These were tested for the presence of TAT together with sera from 20 HIV subjects and a supernatant from HIV-1 infected H9 cells. Further was
  • the above sera were each incubated with 1% Triton X-100 and 1 M NaCl.
  • the sera were then subjected to size fractionation in the range of 5-30 kD, using centrifugation (fractionation) columns from Filtron, cf. were used above.
  • the sera were subjected to a dot blot procedure. For this purpose, they were dripped onto a nitrocellulose membrane and fixed by heat. To block remaining protein binding sites, the nitrocellulose membrane was dissolved in 5% milk powder at room temperature for 2 hours in
  • Tween 20 washed before the signal was developed by a colorimetric reaction (see Fig.). It was found that HIV-TAT can be specifically detected in a body sample using the method according to the invention.

Landscapes

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

Abstract

L'invention concerne un procédé de dépistage du VIH-TAT dans un échantillon prélevé sur le corps, ledit procédé comprenant les étapes suivantes: (a) prélèvement d'un échantillon sur le corps et incubation de l'échantillon avec un détergent et un sel concentré; (b) fractionnement de la taille de l'échantillon de l'étape (a); (c) incubation de l'échantillon de l'étape (b) avec un anticorps anti-VIH-TAT. L'invention concerne en outre une trousse permettant de mettre en ÷uvre ledit procédé.
PCT/DE1996/000641 1995-04-13 1996-04-12 Procede de depistage du vih-tat WO1996032646A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
EP96909957A EP0820595A1 (fr) 1995-04-13 1996-04-12 Procede de depistage du vih-tat
JP8530644A JPH11503524A (ja) 1995-04-13 1996-04-12 Ｈｉｖ−ｔａｔの検出方法

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE1995114089 DE19514089C2 (de) 1995-04-13 1995-04-13 Nachweisverfahren für HIV-TAT
DE19514089.3 1995-04-13

Publications (1)

Publication Number Publication Date
WO1996032646A1 true WO1996032646A1 (fr) 1996-10-17

Family

ID=7759680

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/DE1996/000641 WO1996032646A1 (fr) 1995-04-13 1996-04-12 Procede de depistage du vih-tat

Country Status (4)

Country Link
EP (1) EP0820595A1 (fr)
JP (1) JPH11503524A (fr)
DE (1) DE19514089C2 (fr)
WO (1) WO1996032646A1 (fr)

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1123419B1 (fr) * 1998-08-21 2005-11-09 Neovacs Méthode pour déterminer le pronostic d'individus infectés par le vih
US7067246B1 (en) 1998-08-21 2006-06-27 Neovacs S.A. Method for determining prognosis of HIV infected individuals
FR2792206B1 (fr) * 1999-04-13 2006-08-04 Centre Nat Rech Scient Vaccin anti-vih-1 comprenant tout ou partie de la proteine tat de vih-1
WO2004011948A1 (fr) * 2002-07-25 2004-02-05 Ajinomoto Co., Inc. Methode d'analyse d'une proteine

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0214613A2 (fr) * 1985-09-06 1987-03-18 Bayer Corporation Méthode de détermination d'un comptage différentiel de globules blancs sanguins
BE1001303A6 (nl) * 1987-12-11 1989-09-19 Buyzere Marc Leon Dominique De Enzymatische bepalingsmethode voor creatine in rode bloedcellen na selectieve uitzouting van het hemoglobine.
EP0484765A2 (fr) * 1990-11-06 1992-05-13 Biotest Ag Méthode et moyens de dosage de protéines dans des fluides corporels
WO1992019971A1 (fr) * 1991-04-30 1992-11-12 Alkermes, Inc. Anticorps cationises contre des proteines intracellulaires
EP0557897A1 (fr) * 1992-02-28 1993-09-01 F. Hoffmann-La Roche Ag Protéines liantaux antigènes et procédé de leur production

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE3907562A1 (de) * 1989-03-09 1990-09-13 Bayer Ag Antisense-oligonukleotide zur inhibierung der transaktivatorzielsequenz (tar) und der synthese des transaktivatorproteins (tat) aus hiv-1 und deren verwendung
CA2044679A1 (fr) * 1990-06-22 1991-12-23 Alexander Honigman Depistage de l'infection a vih par bioluminescence

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0214613A2 (fr) * 1985-09-06 1987-03-18 Bayer Corporation Méthode de détermination d'un comptage différentiel de globules blancs sanguins
BE1001303A6 (nl) * 1987-12-11 1989-09-19 Buyzere Marc Leon Dominique De Enzymatische bepalingsmethode voor creatine in rode bloedcellen na selectieve uitzouting van het hemoglobine.
EP0484765A2 (fr) * 1990-11-06 1992-05-13 Biotest Ag Méthode et moyens de dosage de protéines dans des fluides corporels
WO1992019971A1 (fr) * 1991-04-30 1992-11-12 Alkermes, Inc. Anticorps cationises contre des proteines intracellulaires
EP0557897A1 (fr) * 1992-02-28 1993-09-01 F. Hoffmann-La Roche Ag Protéines liantaux antigènes et procédé de leur production

Also Published As

Publication number Publication date
DE19514089A1 (de) 1996-10-24
EP0820595A1 (fr) 1998-01-28
JPH11503524A (ja) 1999-03-26
DE19514089C2 (de) 1997-07-10

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