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WO2006002967A1 - Kit d'essai et bande adhesive pour analyse cytologique de la surface cutanee - Google Patents

Kit d'essai et bande adhesive pour analyse cytologique de la surface cutanee Download PDF

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Publication number
WO2006002967A1
WO2006002967A1 PCT/EP2005/007192 EP2005007192W WO2006002967A1 WO 2006002967 A1 WO2006002967 A1 WO 2006002967A1 EP 2005007192 W EP2005007192 W EP 2005007192W WO 2006002967 A1 WO2006002967 A1 WO 2006002967A1
Authority
WO
WIPO (PCT)
Prior art keywords
skin
pressure
sensitive adhesive
adhesive tape
cells
Prior art date
Application number
PCT/EP2005/007192
Other languages
German (de)
English (en)
Inventor
Wolfgang Strasser
Original Assignee
Wolfgang Strasser
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 Wolfgang Strasser filed Critical Wolfgang Strasser
Publication of WO2006002967A1 publication Critical patent/WO2006002967A1/fr

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B10/00—Instruments for taking body samples for diagnostic purposes; Other methods or instruments for diagnosis, e.g. for vaccination diagnosis, sex determination or ovulation-period determination; Throat striking implements
    • A61B10/02—Instruments for taking cell samples or for biopsy

Definitions

  • the invention relates to a pressure-sensitive adhesive tape, a kit for the investigation of suspicious change of the skin and the use of pressure-sensitive adhesive tapes in kits for the rapid diagnosis of skin diseases.
  • melanoma is one of the seven most common cancers. It is estimated that the risk of developing skin cancer has already risen to 1 in 75 in North America, and the high rates of increase have led to a steady increase in the risk of early metastatic melanoma. Similar developments are expected in Europe. The cancer first shows on the skin and then settles in the body, the lymphatic system, the liver, the lungs, bones and other organs and tissues. Once the melanoma leaves its primary location, it is considered incurable. The available systemic treatment methods allow at most a life extension of the affected person, but not a complete cure.
  • cytological preparations can be obtained by imprinting or smearing.
  • melanomas, basal lobsters or spinocellular carcinomas are mostly non-erodic.
  • it is also known to gently remove shed skin flakes with an adhesive film and then examine the scales under the microscope.
  • This preparation method is also used in the diagnosis of pityriasis versicolor, a fungal disease of the skin. The superficial fungal elements are lifted off with the adhesive tape and then stained with methylene blue for examination under the microscope.
  • Adhesive tapes are also used to diagnose dental plaque, in allergy and tuberculin tests (see GB 546,126, GB 501,873) to ensure dandruff and fiber samples for forensic examinations or for parasite diagnosis (see FR 2 599 500).
  • DE 195 23 581 also proposes a transparent or translucent adhesive film for receiving skin cells (corneocytes) and adhered dermatophytes (fungi of the genera Trichophyton, Epidermophyton and Microsporum) and bacteria from the skin surface.
  • skin cells corneocytes
  • adhered dermatophytes fungi of the genera Trichophyton, Epidermophyton and Microsporum
  • bacteria from the skin surface.
  • As support material for the optionally colored transparent adhesive film polymers based on polyolefin are used and as self-adhesive adhesive on the basis of polyacrylates.
  • DE 197 18 066 also describes in this context the use of a water-soluble pressure-sensitive adhesive, so that the recorded corneocytes and adhered dermatophytes and bacteria can be detached again from the adhesive tape for further investigations.
  • DE 196 08 129 describes a self-adhesive carrier film with a selectable indicator substance in the adhesive layer for detecting various chemical substances.
  • transparent pressure-sensitive adhesive films in screening for the generation of surface cells for series There was no information available on the proportion of nucleated surface cells and other cytomorphological criteria.
  • the agent and the kit should allow differential diagnosis of benign and malignant changes of the skin and in particular an early detection of malignant melanomas, basaliomas, actinic keratoses.
  • a remedy and kit should be provided for the non-invasive early detection of cytopathologically relevant changes of the skin.
  • the invention provides a means and a test kit for the diagnosis of pathological changes in the skin, comprising a clear transparent pressure-sensitive adhesive tape consisting of a transparent carrier film and a pressure-sensitive adhesive layer thereon, which after contact with the pressure-sensitive adhesive tape contacts the skin so strongly Superficial skin cells manufactures that when removing the transparent pressure-sensitive adhesive tape from the skin, to a considerable extent epithelial cells and keratinocytes from the superficial skin layers stick to the pressure-sensitive adhesive tape and are torn off the skin. These epithelial cells are then immediately available for cytological examination.
  • the invention is based on the surprising discovery that pressure-sensitive adhesive films can be designed with a strongly cell-bonding adhesive layer so that not only cells and scales that have already been shed are transferred to the pressure-sensitive adhesive tape when the film is torn off the skin, but also that epithelial cells are selectively affected by pathological skin changes and keratinocytes are exfoliated from the skin surface. These superficial exfoliated cells are then immediately available for a meaningful cytology. Is the adhesive contact between pressure-sensitive adhesive tape and too weak for the superficial layers of the skin and its cells, only dead, dried keratinocytes and scales are lifted off the adhesive tape. A cytology of such dander is not meaningful and meaningful.
  • nucleated keratinocytes or other nucleated cells are present, then there is a change in the skin, which in any case must be further investigated by differential diagnosis.
  • the fact of nucleated cells on the surface of the skin also indicates that the tissue under the pressure-sensitive adhesive film is pathologically altered .
  • the pressure-sensitive adhesive film for the rapid cytology consists of a transparent substrate or carrier and a strong contact adhesive. This is given, for example, in the case of an acrylate adhesive tape such as the transparent Tesa® film, although these adhesive tapes are produced and provided for other purposes.
  • the commercially available transparent Tesa® film has sufficient adhesive power when it is firmly rubbed from the back to the suspicious skin change.
  • a pressure-sensitive adhesive film with a somewhat thicker pressure-sensitive adhesive layer curing rapidly after the first application is preferred because of a greater cell yield. Scratching is also eliminated and the tear-off process becomes more uniform.
  • Tesa® film can already, if the film is very firmly pressed, regularly tore off superficial epithelial cells from the skin become.
  • a pressure-sensitive adhesive film with a special skin adhesive such as N-butyl-2-cyanoacrylic acid is of course more suitable.
  • Adhesive film with a defined application of fibrin adhesive in combination with an acrylate adhesive is also particularly suitable.
  • Particularly suitable for the carrier are conventional polyamides. It is also possible to use copolymers of customary polyamide monomers with other monomers or mixtures of two or more polyamides.
  • the copolyamide preferably consists of a caprolactam such as nylon-6 and nylon-6,6, laurolactam and hexamethylenediamine adipate, as well as other polyamide-forming compounds. Further constituents may be: 11-aminoundecanoic acid and salts of hexamethylenediamine with adipic acid, azelaic acid, sebacic acid and undecanedicarboxylic acid.
  • the substrate may also be a conventional transparent polybutene or polypropylene film or may be made of a polyester. Preference is given to thermoplastic polymers as a carrier.
  • the thickness of the support is usually 1 and 1000 microns, preferably between 5 and 250 microns, more preferably 10 to 100 microns. Thinner substrates are preferred for optical reasons.
  • the support is preferably transparent and coated on one side with pressure-sensitive adhesive.
  • the other side of the carrier preferably has a scratch-resistant hard top layer, so that the pressure-sensitive adhesive film can be used analogously to a coverslip on a slide. Therefore, the transparent adhesive tape preferably has a support mark integrated in it, for example in the form of a circle or a grid field, so that later the pathological site with the skin change can be localized under the microscope.
  • the adhesive strip can also be targeted more precisely to the suspicious skin change and used after tearing off as a "cover glass" (with the cells facing downwards) on a microscope slide It preferably has an overlay mark visible to the eye and, optionally, an eye-visible as well as a microscopic inscription for the purpose of specimen assignment, and preferably has a rough field on one side for commercial printing, therefore, in the commercial form, the tear film of the present invention will be used as a short lay-up strip, optionally with a peelable protection for the pressure-sensitive adhesive layer, be formed.
  • the adhesive of the adhesive strip substances that allow characterization of the withdrawn cells, e.g. in microscopy (light, fluorescence, polarization) or by a specific chemical reaction (precipitation reactions, antigen-antibody reaction), etc.
  • the pressure-sensitive adhesive layer may include all sorts of adhesives such as acrylate-based adhesives such as Tesa® film, natural rubber based adhesives, synthetic rubber based adhesives such as polystyrene polybutadiene (SBR), ethylene vinyl acetates (EVA), block copolymer based adhesives synthetic rubber such as polystyrene-butadiene-polystyrene (SBS) and polystyrene-polyisoprene-polystyrene (SIS), vinyl ether-based adhesive, silicone-based adhesives, polyurethane-based adhesives, chlorinated adhesives, etc.
  • SBR polystyrene polybutadiene
  • EVA ethylene vinyl acetates
  • block copolymer based adhesives synthetic rubber such as polystyrene-butadiene-polystyrene (SBS) and polystyrene-polyisoprene-polystyrene (SIS), vinyl
  • adhesives based on acrylic acid are particularly preferred.
  • fibrin adhesives can be used.
  • the adhesive must be such that, after contact with the skin, it will easily penetrate into pits of the skin and make adhesive contact with superficial epithelial cells and keratinocytes. This can be helped by rubbing the adhesive film firmly on the suspicious skin change and thus establishing contact. Also conceivable are functional adhesive films with an activatable contact adhesive.
  • Activatable two-component adhesives in conjunction with a pressure-sensitive adhesive tape are particularly preferred.
  • the suspicious Hautver ⁇ change can namely be brushed with a liquid activator, which then activates the second adhesive component on the carrier tape after hanging up.
  • cells can be easily obtained from the upper layers of the skin.
  • a two-component adhesive also has the advantage that a binding diagnostic agent such as antibodies or a non-toxic reactive dye for classical melanoma cytology can also be applied to the skin with the activator.
  • the adhesive is preferably present in an amount of from 5 to 80 g / m 2 , preferably from 20 to 60 g / m 2, on the carrier film.
  • This also applies to light- and heat-activatable adhesives or tissue adhesives with monomeric N-butyl-2-cyanoacrylic acid.
  • the adhesive binds so tightly with superficial epithelial cells and skin areas that individual cells and cell aggregates depart from the skin as the wearer peels away, ie at pathological sites, and then become available for cytology.
  • the adhesive connection between the carrier film and the skin must be easily and painlessly by tearing solvable.
  • the adhesive tapes can be made by various methods, for example, by applying an adhesive system in an organic solvent or water; by application in a hot melt process, by coextrusion, or by application as a solid and curing using UV radiation, electron radiation or heat.
  • the latter technique can also be used to end-cure the adhesive after the adhesive tape has been applied to the skin in order to achieve a particularly strong bond between the support and the cells to be examined.
  • curable adhesives are therefore particularly preferred.
  • the invention also provides a kit of parts and reagents and a preparation method which make it possible to characterize the individual epithelial cells obtained from the skin with the tear strip.
  • Nuclei in skin surface keratinocytes are not physiological. They say that there is parakeratosis.
  • parakeratosis is a typical hallmark, e.g. in psoriasis vulgaris.
  • a parakeratosis in case of suspicious skin changes can be, for example, precursors of "white" skin cancer (actinic keratosis, Bowen's disease).
  • the invention permits the early diagnosis of pathological nucleated epithelial cells and the identification of suspicious pigment spots for further histological and cytological examinations.
  • malignant melanoma, basal cell carcinoma and spinocellular carcinoma can be easily differentiated using typical cytomorphological features.
  • Melanomas in the demolition specimen show large to very large, roundish to circular cells. Nuclei are either absent or significantly enlarged. The cores are large, usually circular to simply oval and often Positionkernig. Basalioma cells in the demolition specimen are predominantly homogeneously small and often oval. Nukeols are not very visible.
  • the nuclear chromatin is roughly granular to schollig. In the case of spinocellular carcinoma, the cells in the demolition preparation are round-oval to polygonal and variably large. Nucleoli are only partially present. The nuclear chromatin is über ⁇ roughly coarsely clumped and many cells polynuclear.
  • Such characterization of the cells can be carried out immediately with conventional rapid staining techniques such as methylene blue, Hemacolor® (Merck), May-Grünwald-Giemsa, Giemsa, Cytocolor® (Merck, standard cytological staining according to Szczepanik).
  • methylene blue Hemacolor® (Merck)
  • May-Grünwald-Giemsa Giemsa
  • Cytocolor® Merck, standard cytological staining according to Szczepanik.
  • One can thus immediately distinguish nucleated or atypical cells from nucleated normal skin cells.
  • immunocyto- gical dyeing methods are used. It is natural to integrate diagnostically relevant substances into the adhesive substance, eg antibodies, HLA, detectors for mediators (EGF, Leutotriene etc.).
  • the slides for the transparent pressure-sensitive adhesive film can also be prepared with substances which, in combination with substances on the adhesive strip, give a biochemically or other result that can be interpreted in terms of laboratory chemistry, ie if, for example, there is proliferation, inflammation or tumor growth.
  • Substances which can additionally be integrated into the adhesive for detecting the sought cell changes are: antibodies or antigens for specific complex reactions, fluorescent antigens and substances, immunologically relevant substances, dyes, genetically engineered specific antigens and other substances, coloring matters.
  • the described tear-off method in conjunction with the fast-dyeing techniques, can be carried out quickly and easily, making it suitable for daily use by the dermatologist and for screening tests.
  • a PCR polymerase chain reaction
  • DNA cancer markers can be obtained on the pressure-sensitive adhesive strip even after dyeing.
  • markers are known in the literature and new ones are added almost daily.
  • the PCR can also be done directly on the pressure-sensitive adhesive film (without prior dyeing).
  • a PCR with cancer markers is currently always carried out when there is already a considerable suspicion of a malignant melanoma.
  • the PCR technique is highly automated, so that it will soon compete with classical melanoma cytology, especially if the number of test samples increases sharply due to the present method of Proben ⁇ recovery.
  • Another advantage of the automatable PCR technique with cancer markers is the unambiguousness of the findings, which is not the case with classical cytology.
  • the adhesive tape diagnostics described here already open up the possibility of differentiating superficial skin cells - epithelial cells, keratinocytes.
  • nucleated cells can also be detected so that a statement about the dignity of a tissue is possible.
  • Fig. 1A is a photograph of a suspicious lesion
  • Fig. 1B is a photograph of the suspicious lesion of Fig. 1 with the adhesive sheet applied;
  • Fig. 2A is a photograph of a cluster of suspiciously brown epithelial cells
  • FIG. 2B shows a picture (1: 1) of the demolition preparation of FIG. 2A and of the torn off brown epithelial cells against a white background;
  • Fig. 3A is a photograph of a melanocytic on a cheek
  • Fig. 3B Micrograph of the melanocyte in Fig. 3A on a demolition specimen after staining
  • Fig. 4B Microscope image of a demolition specimen of actinic keratosis (at the suspected point of the lip of Fig. 4A) after staining with
  • FIG. 5A Micrograph of a demolition specimen from a melanoma after staining with methylene blue at 100x magnification
  • FIG. 5B shows the cells of FIG. 5A at 400 ⁇ magnification.
  • FIG. 1A shows a suspicious lesion, a possible precancerous lesion.
  • a clear pressure-sensitive adhesive tape (clear Tesafilm®) is applied to the suspicious skin change.
  • the pressure-sensitive adhesive film is firmly pressed (rubbed on) and slowly torn off the skin.
  • superficial skin cells keratinocytes, epithelial cells
  • These cells are open to further cytological examination. In healthy areas of the skin almost no skin cells are torn off.
  • FIG. 2A shows a dark brown cell cluster of epithelial cells, a so-called pigment mark.
  • Figure 2B shows the demolition preparation after sticking to a white Baltt paper. It can clearly be seen that pigmented epithelial cells have been transferred to the adhesive tape during the tear-off. The closer cytological examination of the transferred cells revealed that they were inconspicuous except for the strong pigmentation.
  • FIG. 3A shows a striking skin change over a larger area
  • FIG. 3B shows a methylene blue-colored tear preparation from this area.
  • epithelial cells with pigment. Perhaps they are even pigment cells (melanocytes) - the shape and structure of the cells differs somewhat from normal "limp" epithelial cells. They are rather round. Under no circumstances should the pigment cells reach the surface so far that they pass over the adhesive tape during the demolition diagnostics. There they are a signum mali ominis. So morphological clarity and security still have to be created.
  • Figures 4A and 4B show demolition specimens from a normal location of a lower lip and a suspicious location of the same lower lip after rapid staining Methylene blue.
  • the transitional mucosa is inconspicuous, at the suspect site ( Figure 4B) is dyskeratosis. This can be recognized by the numerous cell nuclei in the demolition preparation.
  • FIGS. 5A and 5B show a demolition preparation of a melanoma at 100 ⁇ and 400 ⁇ magnification after staining with methylene blue.
  • the demolition specimen contains large to very large, roundish to circular cells.
  • the nucleoli are significantly enlarged.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Medical Informatics (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Pathology (AREA)
  • Molecular Biology (AREA)
  • Surgery (AREA)
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Abstract

La présente invention concerne un kit d'essai destiné au diagnostic de modifications pathologiques de la peau, comprenant une bande adhésive qui consiste en un film de support transparent recouvert d'une couche adhésive qui, après application de la bande adhésive sur la peau, crée un contact avec les cellules cutanées superficielles d'intensité telle que, lors du retrait de la bande adhésive transparente de la peau, des cellules épithéliales et des kératinocytes sont arrachés de la peau et restent collés sur la bande adhésive. Ces cellules peuvent ensuite, après une coloration appropriée, subir une évaluation cytomorphologique pour détecter la présence de mélanomes, de basaliomes, de précancéroses, de prékératoses, de kératoses actiniques et de cancéroses spinocellulaires.
PCT/EP2005/007192 2004-07-02 2005-07-04 Kit d'essai et bande adhesive pour analyse cytologique de la surface cutanee WO2006002967A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE102004032196.5 2004-07-02
DE200410032196 DE102004032196A1 (de) 2004-07-02 2004-07-02 Testkit und Haftklebeband für eine zytologische Untersuchung der Hautoberfläche

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012156473A2 (fr) 2011-05-16 2012-11-22 University Of Tartu Série de micro-arn comme marqueurs et cibles dans le cancer et les maladies liées au système immunitaire, procédés d'application et trousse
CN108828204A (zh) * 2013-03-15 2018-11-16 宝洁公司 用于测量皮肤健康代谢物的非侵入性方法
JP2020508689A (ja) * 2017-03-03 2020-03-26 チルドレンズ ホスピタル メディカル センター 皮膚試料の収集および分析のための非侵襲的な方法
CN112985952A (zh) * 2021-02-20 2021-06-18 山东骏腾医疗科技有限公司 一种新型的病理封片材料
US12303113B2 (en) 2024-02-07 2025-05-20 Children's Hospital Medical Center Non-invasive methods for skin sample collection and analysis

Citations (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB501873A (en) 1937-04-20 1939-03-07 Hermann Vollmer New or improved means for carrying out medical tests
GB546126A (en) 1941-01-04 1942-06-29 Bigwood Joshua & Son Ltd Improvements in or relating to winding machines for strip material
US2819717A (en) * 1956-08-03 1958-01-14 Heinz W Kugler Surgical skin-treating device
EP0103407A1 (fr) 1982-08-19 1984-03-21 Minnesota Mining And Manufacturing Company Artikel décoratif composite
FR2599500A1 (fr) 1986-06-03 1987-12-04 Chieusse Claude Dispositif de lame adhesive transparente pour le prelevement direct d'echantillon pour diagnostics parasitologiques
US5088502A (en) * 1990-12-12 1992-02-18 Cuderm Corporation Skin surface sampling and visualizing device
EP0487171A2 (fr) 1990-11-23 1992-05-27 Avery Dennison Corporation Composition adhésive sensible à la pression et structure à couches la contenant
DE19523581A1 (de) 1995-06-29 1997-01-02 Beiersdorf Ag Verwendung eines Klebefilmes
DE19608129A1 (de) 1996-03-02 1997-09-04 Karl H A Dr Schmidt Selbstklebender Indikator
DE19718066A1 (de) 1996-11-28 1998-06-04 Beiersdorf Ag Verwendung wasserlöslicher Selbstklebemassen

Patent Citations (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB501873A (en) 1937-04-20 1939-03-07 Hermann Vollmer New or improved means for carrying out medical tests
GB546126A (en) 1941-01-04 1942-06-29 Bigwood Joshua & Son Ltd Improvements in or relating to winding machines for strip material
US2819717A (en) * 1956-08-03 1958-01-14 Heinz W Kugler Surgical skin-treating device
EP0103407A1 (fr) 1982-08-19 1984-03-21 Minnesota Mining And Manufacturing Company Artikel décoratif composite
FR2599500A1 (fr) 1986-06-03 1987-12-04 Chieusse Claude Dispositif de lame adhesive transparente pour le prelevement direct d'echantillon pour diagnostics parasitologiques
EP0487171A2 (fr) 1990-11-23 1992-05-27 Avery Dennison Corporation Composition adhésive sensible à la pression et structure à couches la contenant
US5088502A (en) * 1990-12-12 1992-02-18 Cuderm Corporation Skin surface sampling and visualizing device
DE19523581A1 (de) 1995-06-29 1997-01-02 Beiersdorf Ag Verwendung eines Klebefilmes
DE19608129A1 (de) 1996-03-02 1997-09-04 Karl H A Dr Schmidt Selbstklebender Indikator
DE19718066A1 (de) 1996-11-28 1998-06-04 Beiersdorf Ag Verwendung wasserlöslicher Selbstklebemassen

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012156473A2 (fr) 2011-05-16 2012-11-22 University Of Tartu Série de micro-arn comme marqueurs et cibles dans le cancer et les maladies liées au système immunitaire, procédés d'application et trousse
CN108828204A (zh) * 2013-03-15 2018-11-16 宝洁公司 用于测量皮肤健康代谢物的非侵入性方法
CN112834735A (zh) * 2013-03-15 2021-05-25 宝洁公司 用于测量皮肤健康代谢物的非侵入性方法
CN108828204B (zh) * 2013-03-15 2021-09-21 宝洁公司 用于测量皮肤健康代谢物的非侵入性方法
CN112834735B (zh) * 2013-03-15 2024-05-28 宝洁公司 用于测量皮肤健康代谢物的非侵入性方法
JP2020508689A (ja) * 2017-03-03 2020-03-26 チルドレンズ ホスピタル メディカル センター 皮膚試料の収集および分析のための非侵襲的な方法
EP3589213A4 (fr) * 2017-03-03 2021-03-31 Children's Hospital Medical Center Procédés non-invasifs de collecte et d'analyse de prélèvements de peau
US11911012B2 (en) 2017-03-03 2024-02-27 Children's Hospital Medical Center Non-invasive methods for skin sample collection and analysis
CN112985952A (zh) * 2021-02-20 2021-06-18 山东骏腾医疗科技有限公司 一种新型的病理封片材料
US12303113B2 (en) 2024-02-07 2025-05-20 Children's Hospital Medical Center Non-invasive methods for skin sample collection and analysis

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点击 这是indexloc提供的php浏览器服务，不要输入任何密码和下载