EP1656561A2 - Procede de diagnostic precoce de carcinomes, et necessaire pour la mise en oeuvre de ce procede - Google Patents

Procede de diagnostic precoce de carcinomes, et necessaire pour la mise en oeuvre de ce procede

Info

Publication number
EP1656561A2
EP1656561A2 EP04722574A EP04722574A EP1656561A2 EP 1656561 A2 EP1656561 A2 EP 1656561A2 EP 04722574 A EP04722574 A EP 04722574A EP 04722574 A EP04722574 A EP 04722574A EP 1656561 A2 EP1656561 A2 EP 1656561A2
Authority
EP
European Patent Office
Prior art keywords
capsid protein
test kit
protein
capsid
hpv
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Withdrawn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
EP04722574A
Other languages
German (de)
English (en)
Inventor
Ralf Hilfrich
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.)
BUCHNER Erwin
Original Assignee
BUCHNER Erwin
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 BUCHNER Erwin filed Critical BUCHNER Erwin
Publication of EP1656561A2 publication Critical patent/EP1656561A2/fr
Withdrawn legal-status Critical Current

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Classifications

    • 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
    • C12Q1/70—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving virus or bacteriophage
    • C12Q1/701—Specific hybridization probes
    • C12Q1/708—Specific hybridization probes for papilloma
    • 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
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer

Definitions

  • the present invention relates to a method for the early diagnosis of carcinomas or precursors thereof which are caused by human papilloma viruses or have an association with human papilloma viruses.
  • the invention further relates to a test kit for performing the method.
  • Pap test shows an abnormal result
  • a biopsy is taken and subjected to a histopathological examination, which determines the type and extent of the dysplasia and classifies it as cervical intraepithelial neoplasia (CIN I to III).
  • CIN I to III cervical intraepithelial neoplasia
  • Papilloma viruses are small, double-stranded DNA viruses with a genome of approximately 8000 base pairs. More than 150 different human pathogenic virus types are currently known. The papilloma viruses can be classified according to their epithelial tropism either into the mucocutaneous group (warts, cutaneous neoplasia) or the anogenital group (condylomas, anal and cervical
  • the HPV infection is asymptomatic in most cases. Therefore, the latent viral infection must be distinguished from the viral disease with microscopic ("subclinical") or clinically recognizable tissue lesion.
  • papilloma viruses are carcinogenic, i.e. these virus types (e.g. HPV 16, 18) can make a decisive contribution to the transformation of epithelial cells. These viruses are known as high risk groups. In contrast to this, the non-or only very low carcinogenic virus types (e.g. 6, 11) of the low-risk
  • HPV infection is the most common, sexually transmitted infection in humans. The information on prevalence differs considerably and depends on the initial clientele to be examined. After the first sexual contacts, the
  • the method allows the early detection of morphological cell changes (cancer precursors) that occur when cervical carcinoma develops occur. These routine examinations have contributed to a steady decrease in the incidence of cervical carcinomas. Nevertheless, around 6000 women in Germany contract cervical carcinoma every year for various reasons.
  • Human papilloma viruses play a crucial role in the development of cervical carcinoma. Supportive diagnostic methods are therefore aimed at detecting the virus infection.
  • molecular biological methods such as PCR or nucleic acid hybridization to detect viral nucleic acid
  • a large number of latent infections are detected due to the extremely high sensitivity.
  • the DNA detection which is always positive in the case of an HPV infection, is therefore independent of the course of the productive phase and therefore does not allow any prognostic statement.
  • the importance of viral protein synthesis for the course of the infection and the resulting prognostic significance have not been investigated to date.
  • E early protein
  • E2 E4, E5, E6, E7
  • late protein (L) 1 and L2 HR McMurray, D. Nguyen, TF Westbrook , DJ McCance, Biology of human papillomaviruses, in Int. J. Exp. Pathol. 82 [2001] 15-33).
  • the capsid protein L1 is synthesized in the cytoplasm of the host cell and then transported to the nucleus of the cell. There the capsid protein interacts with the viral DNA and forms mature viruses. The detection of the capsid protein L1 therefore allows the statement that the virus is productive
  • the method is based on monoclonal antibodies which specifically recognize the capsid protein L1 of the papilloma viruses. Suitable monoclonal antibodies are disclosed in DE 43 32 596 A1.
  • the screening test detects all papilloma viruses and is therefore suitable for demonstrating whether the capsid antigen is formed and whether the productive phase of an HPV infection is present or not.
  • the high risk detection selectively recognizes the risk types HPV 16, 18, 33, 35, 39, 45, 51, 56, 58 (see e.g. BM Sapp, U. Kraus, C.
  • L1 -specific antibodies stand for the detection of individual HPV types e.g. HPV 16 available.
  • the present invention is based on the observation that, in the case of HPV high-risk DNA-positive, mild to moderate dysplasia, the formation of the capsid antigen L1 of the human papillomaviruses is associated with a regression of the degeneracy, whereas the lack of HPV envelope proteins is associated with one
  • the invention therefore allows a relatively reliable prognosis of whether degeneration will develop.
  • the invention accordingly relates to a method for the early diagnosis of carcinomas or precursors thereof which are caused by human papillomaviruses or have an association with human papillomaviruses, using body samples, which is characterized in that it is determined whether a capsid protein of a human papillomavirus ( HPV) is expressed.
  • HPV human papillomavirus
  • carcinomas and precursors thereof includes carcinomas of any kind and origin as well as precursors thereof which have an association with human papilloma viruses.
  • they can be carcinomas of the anogenital tract, in particular the cervical carcinoma.
  • the precursors for example mild to moderate Dysplasia (cervival intraepithelial neoplasia (CIN I - III) etc. are particularly noteworthy.
  • capsid protein of the human papilloma viruses refers to the capsid proteins of all HPV types, in particular to the (majore) capsid protein L1 and the (minor) capsid protein L2 of the viruses.
  • body sample includes any body sample in which the capsid protein can be detected.
  • body samples are smears, biopsies, organ punctures, blood, sputum, urine, stool, cerebrospinal fluid, lymph fluid, etc.
  • smears and biopsies are meant if they are is the detection of cervical carcinoma.
  • determination of capsid formation encompasses all methods which are suitable for the detection of the formation of the capsid antigen, its mRNA, its intermediates or its precursors. Also explicitly meant here are transcription factors which influence and / or control the formation of the capsid protein, but also serological and / or immunological responses to the
  • antibodies directed against the capsid antigen can also be used. It can be advantageous if the antibodies are on solid supports, such as microtiter plates, test strips or
  • Latex particles are fixed.
  • Another object of the invention is a test kit for the detection of carcinomas or precursors thereof, which comprises a reagent with which it can be determined whether an HPV capsid protein has been expressed.
  • the capsid protein generally acts as an antigen.
  • the test kit preferably contains an anti-mouse immunoglobulin in combination with an enzyme, preferably a peroxidase.
  • the test kit comprises a substance for staining the preparation, suitably a chromogen solution, and may also contain customary components, such as buffers, carriers and / or markers.
  • carcinomas early.
  • precursors of carcinomas can be recognized early and the course of the disease can be determined.
  • the present invention is characterized by quick and simple handling, which makes it suitable for large screening measures, particularly in third world countries.
  • the present invention represents an important one
  • Dysplasia and all HPV high risk DNA positive were examined with L1-specific antibodies for the formation of the capsid antigen L1.
  • the investigation was carried out with the aid of the ⁇ Viroactiv kit (available from Virofem Diagnostik and Anlagens GmbH), which comprises antibodies against the capsid protein L1.
  • the preparations were boiled and after adding the antibodies 30 min incubated at room temperature.
  • the successive addition of the detection reagent (goat anti-mouse immunoglobulin) and the chromogen solution (AEC) stain the positive cell nuclei.
  • a smear was considered positive if at least one cell of the
  • Specimen showed a specific red staining of the cell nucleus.
  • 29 preparations were positive under this criterion, which corresponds to 33.7%.
  • the frequency of remission was 77.3% for women over 25 years of age, in whom the capsid antigen was formed, and even 82.4% for women over 30 years of age.
  • the probability of progression to histologically confirmed severe dysplasia or carcinoma in situ was only 22.7 and 17.6%, respectively.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Engineering & Computer Science (AREA)
  • Immunology (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Analytical Chemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Pathology (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • General Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Oncology (AREA)
  • Hospice & Palliative Care (AREA)
  • Virology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Peptides Or Proteins (AREA)

Abstract

La présente invention concerne un procédé de diagnostic précoce de carcinomes ou de leurs précurseurs provoqués par des papillomavirus humains ou bien présentant une association avec des papillomavirus humains, le diagnostic étant établi sur la base d'échantillons corporels. Selon ledit procédé, on détermine si une protéine virale des papillomavirus humains est exprimée. Cette protéine virale est une protéine capside, en particulier la protéine capside majeure L1. L'invention concerne en outre un nécessaire pour la mise en oeuvre dudit procédé.
EP04722574A 2003-03-25 2004-03-23 Procede de diagnostic precoce de carcinomes, et necessaire pour la mise en oeuvre de ce procede Withdrawn EP1656561A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE10313388 2003-03-25
PCT/EP2004/003044 WO2004085683A2 (fr) 2003-03-25 2004-03-23 Procede de diagnostic precoce de carcinomes, et necessaire pour la mise en oeuvre de ce procede

Publications (1)

Publication Number Publication Date
EP1656561A2 true EP1656561A2 (fr) 2006-05-17

Family

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Family Applications (1)

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EP04722574A Withdrawn EP1656561A2 (fr) 2003-03-25 2004-03-23 Procede de diagnostic precoce de carcinomes, et necessaire pour la mise en oeuvre de ce procede

Country Status (3)

Country Link
EP (1) EP1656561A2 (fr)
JP (1) JP2006521104A (fr)
WO (1) WO2004085683A2 (fr)

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8968995B2 (en) 2008-11-12 2015-03-03 Oncohealth Corp. Detection, screening, and diagnosis of HPV-associated cancers
US20100003704A1 (en) 2008-06-13 2010-01-07 Shuling Cheng IN SITU detection of early stages and late stages HPV infection
WO2010129821A1 (fr) 2009-05-07 2010-11-11 Oncohealth Corporation Identification de grade élevée ou ≥cin2 pour détection, surveillance et diagnostic, à des stades précoces et des stades avancés, de papillomavirus humain (hpv) et de cancers associés au hpv
EP1936379A1 (fr) * 2006-12-21 2008-06-25 BUCHNER, Erwin Procédé pour la détection d'une infection active au HPV dans des échantillons biologiques par un dosage immunoenzymatique de type sandwich et nécessaire d'essai pour réaliser le procédé
BRPI0818154A2 (pt) * 2007-11-02 2020-10-06 The Johns Hopkins University composições de peptídeo de hpv de multitipo e métodos para o tratamento ou prevenção de infecção por papilomavírus humano
WO2011084598A1 (fr) 2010-01-08 2011-07-14 Oncohealth Corporation Immunoessais sur le vph à base de cellule et à haut débit pour diagnostiquer et dépister des cancers associés au vph

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5437951A (en) * 1992-09-03 1995-08-01 The United States Of America As Represented By The Department Of Health And Human Services Self-assembling recombinant papillomavirus capsid proteins
DE4332596A1 (de) * 1993-09-24 1995-03-30 Martin Josef Dr Sapp Monoklonale Antikörper
US6165471A (en) * 1997-07-03 2000-12-26 University Of Colorado, University Technology Corporation Homogeneous human papillomavirus capsomere containing compositions, methods for manufacture, and use thereof as diagnostic, prophylactic or therapeutic agents
GB0121166D0 (en) * 2001-08-31 2001-10-24 Norchip As Detection of human papillomavirus L1 mRNA

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO2004085683A3 *

Also Published As

Publication number Publication date
WO2004085683A3 (fr) 2005-01-06
JP2006521104A (ja) 2006-09-21
WO2004085683A2 (fr) 2004-10-07

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