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WO1999036775A1 - Method for detecting the origin of livestock and the products obtained therefrom - Google Patents

Method for detecting the origin of livestock and the products obtained therefrom Download PDF

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Publication number
WO1999036775A1
WO1999036775A1 PCT/DE1998/000113 DE9800113W WO9936775A1 WO 1999036775 A1 WO1999036775 A1 WO 1999036775A1 DE 9800113 W DE9800113 W DE 9800113W WO 9936775 A1 WO9936775 A1 WO 9936775A1
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WIPO (PCT)
Prior art keywords
immunogen
livestock
farm animal
enzyme
use according
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.)
Ceased
Application number
PCT/DE1998/000113
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German (de)
French (fr)
Inventor
Wolf Bertling
Manfred Gareis
Martin Groschup
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.)
November AG Novus Medicatus Bertling Gesellschaft fuer Molekular Medizin
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November AG Novus Medicatus Bertling Gesellschaft fuer Molekular Medizin
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Application filed by November AG Novus Medicatus Bertling Gesellschaft fuer Molekular Medizin filed Critical November AG Novus Medicatus Bertling Gesellschaft fuer Molekular Medizin
Priority to EP98905259A priority Critical patent/EP1047940A1/en
Priority to PCT/DE1998/000113 priority patent/WO1999036775A1/en
Priority to CA002315023A priority patent/CA2315023A1/en
Priority to JP2000540436A priority patent/JP2002509251A/en
Publication of WO1999036775A1 publication Critical patent/WO1999036775A1/en
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
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K11/00—Marking of animals

Definitions

  • the invention relates to a method and a use for verifying the origin of farm animals, in particular cattle, pigs and the like. the like , as well as products from it.
  • livestock such as cattle and pigs
  • an identifier for proof of origin which is stamped on a tag attached to the ear of the livestock (tattoo, transponder, etc. systems).
  • the farm animals are grouped according to their origin.
  • the meat of the slaughtered animals is then given a group-specific stamp.
  • the company has started to produce accompanying documents for every farm animal.
  • the proof of origin used according to the prior art is insufficient.
  • the object of the invention is to eliminate the disadvantages of the prior art.
  • forgery-proof proof is to be provided, with which the origin of both the live farm animal and products derived from it, such as meat, meat products and food of animal origin, can be reliably verified.
  • the method according to the invention has the advantage that the biological marking is inextricably linked to the farm animal and therefore cannot be exchanged or counterfeited. It is also possible to prove the origin of meat without a stamp of origin, even of meat used in the manufacture of meat products.
  • a protein and / or peptide is / are advantageously used as the immunogen (s), which / s neither as a vaccine nor via the food chain, nor via the environment in which usual livestock farming gets into the organism of the livestock and against which no antibodies are naturally formed in the livestock.
  • a protein and / or peptide coupled with a carrier is expediently used. - This enables a specific marking to be achieved in a simple manner, which is easily detectable and completely harmless for human consumption.
  • the immunogen is designed in such a way that the specific antibodies formed remain permanently in the organism of the livestock and can be detected by means of an enzyme-immunological or immunochemical detection method.
  • the marking can be carried out by a single application, which is expediently carried out at the earliest possible time after the birth of the farm animal.
  • the application can take place by the immunogen being applied to the farm animal by injection.
  • the immunogen can also pass through the mucous membrane, preferably the nasal mucosa, of the farm animal, e.g. with a nasal spray or by means of an implant.
  • a mixture of several different immunogens is advantageously applied.
  • a first immunogen can be used to identify the region of origin, such as Western Europe, Eastern Europe, South America, etc.
  • a second immunogen can be used to identify the country of origin
  • a third immunogen can be used to identify administrative units in the country concerned.
  • individual production and sales organizations eg producer rings, quality meat programs
  • the DNA coding for it can also be applied with a suitable expression vector. In this way, the immunogen is repeatedly re-expressed. In this case, repeated application of the immunogen to refresh the label is unnecessary.
  • Blood, milk or other body secretions can be used to detect the antibodies in the live farm animal, and blood and body secretions such as the dripping or squeezed juice of the meat can also be used in the slaughtered animal.
  • enzyme immunological or immunochemical detection is particularly simple if it is carried out using test sticks. - If the marking remains reserved for veterinarians under state supervision, the origin of e.g. products obtained from slaughtered livestock are practically checked and ascertained by everyone.
  • the immunogen can be a protein and / or peptide or a mixture of these which does not enter the organism of the livestock either as a vaccine or as a medicament, nor via the food chain in normal livestock farming and against which no antibodies are naturally formed in the livestock ; the protein and / or peptide can be coupled to a carrier.
  • an immunogen which is foreign to the organism to be labeled but which is harmless for biological labeling enables a surprisingly simple, inexpensive and forgery-proof proof of origin.
  • the DNA coding for it can also be applied.
  • the corresponding immunogen is formed in the body of the farm animal by expression. This ensures permanent marking.
  • the farm-animal can immunogens such as "keyhole limpet hemocyanin", "green fluorescent protein” are administered from Aequoria victoria, inactive snake toxins and Viruspro ⁇ teine.
  • immunogens such as "keyhole limpet hemocyanin", "green fluorescent protein” are administered from Aequoria victoria, inactive snake toxins and Viruspro ⁇ teine.
  • naturally occurring peptides or polypeptides such as hirudin, pheromonotropin or ranalexin can also be applied.
  • synthetically produced fragments of partial amino acid sequences of naturally occurring immunogens can also be used for biomarking.
  • artificial proteins and peptides, their amino acid sequence can also serve as immunogens does not correspond to any previously known substance, but is only assembled or manufactured for the purpose of biomarking
  • a DNA can also be applied which has a nucleotide sequence coding for the immunogen.
  • the DNA can contain partial sequences or the entire gene of "keyhole limpet hemocyanine". However, it can also be designed such that it describes only a short peptide, it being possible for several immunogens to be encoded on a single DNA. In the case of naturally occurring genes or sequences derived therefrom, both cDNA and genomic DNA with intron sequences can be applied.
  • the DNA can contain further sequences which are important for the expression and processing of the immunogen in the livestock organism.
  • promoter sequences and sequences which bring about transcription termination and polyadenylation of the mRNA and intracellular "trafficking" of the immunogen.
  • Suitable expression regulators are promoters which ensure a high transcription rate of the DNA, e.g. the "immediate early" promoter of the cytomegalovirus or the promoter of a livestock-specific "house-keeping" gene such as ⁇ -actin.
  • promoters are also suitable which specifically restrict the expression of the immunogen to predetermined body cells.
  • the polypeptide formed can be directed into the lumen of the endoplasmic reticulum by suitable "trafficking" signal sequences and directed into cellular distribution channels for secretion or expression and presentation on the cell surface.
  • the antibodies induced by the substances mentioned are detected by common detection methods such as ELISA, EIA and RIA, preferably using test sticks, e.g. be immersed in the dripping juice of the meat of the slaughtered farm animal.

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  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Immunology (AREA)
  • Hematology (AREA)
  • Environmental Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Urology & Nephrology (AREA)
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  • Biotechnology (AREA)
  • Physics & Mathematics (AREA)
  • Birds (AREA)
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  • Zoology (AREA)
  • Animal Husbandry (AREA)
  • Biodiversity & Conservation Biology (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Cell Biology (AREA)
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Abstract

The invention relates to a method for detecting the origin of livestock, especially cattle, swine or the like, comprising the following steps: a) biological marking the live animal by applying at least one antigen that is harmless both for the animal and for human beings and b) identifying the specific antibodies formed by the application of the at least one antigen using an immuno-enzymatic or immuno-chemical detection technique.

Description

Verfahren zum Herkunftsnachweis von Nutztieren sowie davon stammenden ProduktenProof of origin of farm animals and products derived from them

Die Erfindung betrifft ein Verfahren und eine Verwendung zum Herkunftsnachweis von Nutztieren, insbesondere von Rindern, Schweinen u. dgl . , sowie davon stammenden Produkten.The invention relates to a method and a use for verifying the origin of farm animals, in particular cattle, pigs and the like. the like , as well as products from it.

Nach dem Stand der Technik werden Nutztiere, wie Rinder und Schweine, zum Herkunftsnachweis mit einer Kennung versehen, die in eine am Ohr des Nutztiers befestigte Kennmarke eingeprägt ist (Tätowierung, Transponder u.a. Systeme). Vor dem Schlachten werden die Nutztiere in Abhängigkeit ihrer Herkunft in Gruppen zusammengefaßt. Das Fleisch der geschlachteten Tiere wird sodann mit einem gruppenspezifischen Stempel versehen. - Neuerdings ist man zur Erhöhung der Zuverlässigkeit des Herkunftsnachweises dazu übergegangen, zusätzlich zu jedem Nutztier Begleitpapiere zu erstellen.According to the state of the art, livestock, such as cattle and pigs, are provided with an identifier for proof of origin, which is stamped on a tag attached to the ear of the livestock (tattoo, transponder, etc. systems). Before slaughter, the farm animals are grouped according to their origin. The meat of the slaughtered animals is then given a group-specific stamp. - Recently, in order to increase the reliability of the proof of origin, the company has started to produce accompanying documents for every farm animal.

Dennoch sind die nach dem Stand der Technik derzeit verwende- ten Herkunftsnachweise in vielerlei Hinsicht nachteilig. So ist es Gang und Gebe, daß zum Unterlaufen von Importbeschränkungen oder -sperren die Kennmarken ausgetauscht oder gefälscht und die entsprechenden Begleitpapiere ebenfalls ge¬ fälscht werden. Das Erstellen von Begleitpapieren verursacht einen hohen bürokratischen Aufwand. Außerdem sind die herkömmlichen Herkunftsnachweise nicht dauerhaft.Nevertheless, the guarantees of origin currently used according to the state of the art are disadvantageous in many respects. Thus, it is walk and add that exchanged about undermining import restrictions or turnstile stops the characteristic marks or falsified and the corresponding accompanying documents also ge ¬ be deflected. The creation of accompanying documents causes a lot of bureaucratic effort. In addition, the traditional proof of origin is not permanent.

Gerade zur zuverlässigen Eindämmung des illegalen Handelns mit bsp. potentiell BSE-Erreger kontaminierten Rindfleisch bzw. dessen Verwendung zur Herstellung von Fleischprodukten, wie Hundefutter u.a., ist der nach dem Stand der Technik ver¬ wendete Herkunftsnachweis unzureichend. Aufgabe der Erfindung ist es, die Nachteile nach dem Stand der Technik zu beseitigen. Insbesondere soll ein fälschungssicherer Nachweis bereitgestellt werden, mit dem die Herkunft sowohl des lebenden Nutztiers als auch davon stammender Pro- dukte, wie Fleisch, Fleischerzeugnisse und Lebensmittel tierischer Herkunft, zuverlässig nachweisbar sind.Especially for the reliable containment of illegal actions with e.g. Beef potentially contaminated with BSE or its use for the production of meat products, such as dog food, etc., the proof of origin used according to the prior art is insufficient. The object of the invention is to eliminate the disadvantages of the prior art. In particular, forgery-proof proof is to be provided, with which the origin of both the live farm animal and products derived from it, such as meat, meat products and food of animal origin, can be reliably verified.

Diese Aufgabe wird durch die Merkmale der Ansprüche 1 und 15 gelöst. Zweckmäßige Weiterbildungen ergeben sich aus den Merkmalen der Ansprüche 2 bis 14 bzw. 16 bis 25.This object is solved by the features of claims 1 and 15. Appropriate further developments result from the features of claims 2 to 14 and 16 to 25.

Zur Lösung der Aufgabe ist ein Verfahren vorgesehen, welches die folgenden Schritte umfaßt:To achieve the object, a method is provided which comprises the following steps:

a) Biologische Markierung des lebenden Nutztiers durch Applikation mindestens eines für das Nutztier und den Menschen unschädlichen Immunogens, unda) biological labeling of the live farm animal by application of at least one immunogen which is harmless to the farm animal and to humans, and

b) Nachweis der durch die Applikation von mindestens einem Immunogen gebildeten spezifischen Antikörper im lebenden Nutztier oder in davon stammenden Produkten mittels eines enzymimmunologischen oder immunochemisehen Nachweisverfahrens.b) Detection of the specific antibodies formed by the application of at least one immunogen in the live farm animal or in products derived therefrom by means of an enzyme-immunological or immunochemical detection method.

Das erfindungsgemäße Verfahren hat den Vorteil, daß die bio- logische Markierung untrennbar mit dem Nutztier verbunden und daher nicht ausgetauscht oder gefälscht werden kann. Außerdem ist es möglich, die Herkunft von Fleisch ohne Herkunftsstempel, sogar von zur Herstellung von Fleischprodukten verwendetem Fleisch nachzuweisen.The method according to the invention has the advantage that the biological marking is inextricably linked to the farm animal and therefore cannot be exchanged or counterfeited. It is also possible to prove the origin of meat without a stamp of origin, even of meat used in the manufacture of meat products.

Vorteilhafterweise wird/werden als Immunogen (e) ein Protein und/oder Peptid verwendet, das/die weder als Impfstoff noch noch über die Nahrungskette, noch über die Umwelt, bei der üblichen Nutztierhaltung in den Organismus des Nutztiers gelangt/gelangen und gegen das/die in den Nutztieren natürlicherweise keine Antikörper gebildet werden. Dabei wird zweckmäßigerweise ein mit einem Trägerstoff gekoppeltes Protein und/oder Peptid verwendet. - Damit kann auf einfache Weise eine spezifische Markierung erzielt werden, die leicht nachweisbar und für den menschlichen Verzehr völlig unbedenklich ist.A protein and / or peptide is / are advantageously used as the immunogen (s), which / s neither as a vaccine nor via the food chain, nor via the environment in which usual livestock farming gets into the organism of the livestock and against which no antibodies are naturally formed in the livestock. A protein and / or peptide coupled with a carrier is expediently used. - This enables a specific marking to be achieved in a simple manner, which is easily detectable and completely harmless for human consumption.

Nach einem Ausgestaltungsmerkmal ist das Immunogen derart beschaffen, daß die gebildeten spezifischen Antikörper dauerhaft im Organismus des Nutztiers verbleiben und mittels eines enzymimmunologischen oder immunochemisehen Nachweisverfahrens nachweisbar sind. - Auf diese Weise kann die Markierung durch eine einzige Applikation durchgeführt werden, die zweckmäßigerweise zum frühest möglichen Zeitpunkt nach der Geburt des Nutztiers erfolgt. Die Applikation kann erfolgen, indem das Immunogen dem Nutztier durch Injektion appliziert wird. Daneben kann das Immunogen auch über die Schleimhaut, vorzugswei- se die Nasenschleimhaut, des Nutztiers, bsp. mit einem Nasenspray, oder mittels eines Implantats appliziert werden.According to one design feature, the immunogen is designed in such a way that the specific antibodies formed remain permanently in the organism of the livestock and can be detected by means of an enzyme-immunological or immunochemical detection method. In this way, the marking can be carried out by a single application, which is expediently carried out at the earliest possible time after the birth of the farm animal. The application can take place by the immunogen being applied to the farm animal by injection. In addition, the immunogen can also pass through the mucous membrane, preferably the nasal mucosa, of the farm animal, e.g. with a nasal spray or by means of an implant.

Vorteilhafterweise wird eine Mischung von mehreren verschiedenen Immunogenen appliziert. Dabei kann ein erstes Immunogen zur Kennzeichnung der Herkunftsregion, wie Westeuropa, Osteuropa, Südamerika usw., ein zweites Immunogen zur Kennzeichnung des Herkunftsstaats und ein drittes Immunogen zur Kennzeichnung von Verwaltungseinheiten des betreffenden Staats verwendet werden. Alternativ oder ergänzend können auch ein- zelne Produktions- und Vertriebsorganisationen (z.B. Erzeugerringe, Qualitätsfleischprogramme) eine biologische Markierung ihrer Nutztierbestände bzw. davon stammender Produkte anwenden. Anstelle des Immunogens kann auch die dafür kodierende DNA mit einem geeigneten Expressionsvektor appliziert werden. Auf diese Weise wird das Immunogen immer wieder neu exprimiert. Eine wiederholte Applikation des Immunogens zur Auffrischung der Markierung erübrigt sich in diesem Fall.A mixture of several different immunogens is advantageously applied. A first immunogen can be used to identify the region of origin, such as Western Europe, Eastern Europe, South America, etc., a second immunogen can be used to identify the country of origin and a third immunogen can be used to identify administrative units in the country concerned. As an alternative or in addition, individual production and sales organizations (eg producer rings, quality meat programs) can also use a biological label for their livestock or their products. Instead of the immunogen, the DNA coding for it can also be applied with a suitable expression vector. In this way, the immunogen is repeatedly re-expressed. In this case, repeated application of the immunogen to refresh the label is unnecessary.

Zum Nachweis der Antikörper können beim lebenden Nutztier das Blut, die Milch oder andere Körpersekrete, beim geschlachte- ten Tier ebenfalls Blut und Körpersekrete, wie der Tropfoder Pressaft des Fleisches, verwendet werden.Blood, milk or other body secretions can be used to detect the antibodies in the live farm animal, and blood and body secretions such as the dripping or squeezed juice of the meat can also be used in the slaughtered animal.

Zur Identifikation der Antikörper wird als enzymimmunologisches oder immunochemisches vorzugsweise eines oder mehrere der folgenden Nachweisverfahren angewendet: Enzyme Linked Im- munosorbent Assay (= ELISA) , Enzyme Immuno Assay (= EIA) und Radio Immuno Assay (= RIA) . Besonders einfach ist der enzymimmunologische oder immunochemische Nachweis, wenn er unter Verwendung von Teststäbchen durchgeführt wird. - Wenn die Markierung unter staatlicher Aufsicht stehenden Tierärzten vorbehalten bleibt, kann die Herkunft von bsp. aus geschlachteten Nutztieren gewonnenen Produkten praktisch durch jedermann geprüft und festgestellt werden.To identify the antibodies, one or more of the following detection methods are preferably used as enzyme immunological or immunochemical: enzyme linked immunosorbent assay (= ELISA), enzyme immuno assay (= EIA) and radio immuno assay (= RIA). Enzyme immunological or immunochemical detection is particularly simple if it is carried out using test sticks. - If the marking remains reserved for veterinarians under state supervision, the origin of e.g. products obtained from slaughtered livestock are practically checked and ascertained by everyone.

Nach Maßgabe der Erfindung ist des weiteren die Verwendung mindestens einesAccording to the invention, the use of at least one is also

a) für Nutztiere und den Menschen unschädlichen Immunogens zur biologischen Markierung des lebenden Nutztiers sowie da- von stammender Produkte, unda) for livestock and human-harmless immunogen for the biological labeling of live livestock and products derived therefrom, and

b) eines enzymimmunologischen oder immunochemischen Nachweisverfahrens zum Nachweis der durch die Applikation des mindestens einen Immunogens gebildeten spezifischen Antikörper im lebenden Nutztier oder in davon stammenden Produkten vorgesehen.b) an enzyme immunological or immunochemical detection method for the detection of the by the application of at least one specific antibody formed in the live farm animal or in products derived therefrom.

Dabei kann das Immunogen ein Protein und/oder Peptid oder ein Gemisch aus diesen sein, das weder als Impfstoff noch als Arzneimittel, noch über die Nahrungskette bei der üblichen Nutztierhaltung in den Organismus des Nutztiers gelangt und gegen das in den Nutztieren natürlicherweise keine Antikörper gebildet werden; das Protein und/oder Peptid kann mit einem Trägerstoff gekoppelt sein. - Die Verwendung eines dem zu markierenden Organismus fremden, aber unschädlichen Immunogens zur biologischen Markierung erlaubt einen überraschend einfachen, kostengünstigen und fälschungssicheren Herkunfts- nachweis.The immunogen can be a protein and / or peptide or a mixture of these which does not enter the organism of the livestock either as a vaccine or as a medicament, nor via the food chain in normal livestock farming and against which no antibodies are naturally formed in the livestock ; the protein and / or peptide can be coupled to a carrier. - The use of an immunogen which is foreign to the organism to be labeled but which is harmless for biological labeling enables a surprisingly simple, inexpensive and forgery-proof proof of origin.

Anstelle des Immunogens kann auch die dafür kodierende DNA appliziert werden. Das entsprechende Immunogen wird im Körper des Nutztiers durch Expression gebildet. Das gewährleistet eine dauerhafte Markierung.Instead of the immunogen, the DNA coding for it can also be applied. The corresponding immunogen is formed in the body of the farm animal by expression. This ensures permanent marking.

Beispiele :Examples:

Als Proteine, die eine ausgezeichnete spezifische Antikörperbildung stimulieren, können dem Nutztier Immunogene wie "keyhole limpet hemocyanine" , "green fluorescent protein" aus Aequoria victoria, inaktive Schlangentoxine sowie Viruspro¬ teine verabreicht werden. Daneben können auch natürlich vorkommende Peptide oder Polypeptide wie Hirudin, Pheromonotro- pin oder Ranalexin appliziert werden. Des weiteren können zur Biomarkierung auch synthetisch hergestellte Fragmente von Am- minosäureteilsequenzen natürlich vorkommender Immunogene verwendet werden. Schließlich können als Immunogene auch artifi- zielle Proteine und Peptide dienen, deren Amminosäuresequenz keinem bisher bekannten Stoff entspricht, sondern ausschließlich zum Zweck der Biomarkierung zusammen- bzw. hergestellt werden.As proteins that stimulate an excellent specific antibody formation, the farm-animal can immunogens such as "keyhole limpet hemocyanin", "green fluorescent protein" are administered from Aequoria victoria, inactive snake toxins and Viruspro ¬ teine. In addition, naturally occurring peptides or polypeptides such as hirudin, pheromonotropin or ranalexin can also be applied. Furthermore, synthetically produced fragments of partial amino acid sequences of naturally occurring immunogens can also be used for biomarking. Finally, artificial proteins and peptides, their amino acid sequence, can also serve as immunogens does not correspond to any previously known substance, but is only assembled or manufactured for the purpose of biomarking

Zur Markierung kann auch eine DNA appliziert werden, welche eine das Immunogen kodierende Nukleotidfolge aufweist. Die DNA kann Teilsequenzen oder das gesamte Gen von "keyhole lim- pet hemocyanine" enthalten. Sie kann aber auch so ausgebildet sein, daß sie nur ein kurzes Peptid beschreibt, wobei mehrere Immunogene auf einer einzigen DNA kodiert sein können. Im Fall natürlich vorkommender Gene oder davon abgeleiteter Sequenzen kann sowohl cDNA als auch genomische DNA mit Intron- sequenzen appliziert werden. Neben den das Immunogen kodierenden Nukleotiden kann die DNA weitere Sequenzen enthalten, die für die Expression und Prozessierung des Immunogens im Nutztierorganismus von Bedeutung sind. Dabei handelt es sich um Promotorsequenzen sowie um Sequenzen, die eine Transkrip- tionstermination und Polyadenylierung der mRNA sowie ein intrazelluläres "Trafficking" des Immunogens bewirken. Als Ex- pressionsregulatoren eigenen sich Promotoren, die eine hohe Transkriptionsrate der DNA gewährleisten, z.B. der "immediate early" Promotor des Cytomegalovirus oder der Promotor eines nutztierspezifischen "house-keeping" Gens wie ß-Aktin. Es sind aber auch Promotoren geeignet, welche die Expression des Immunogens gezielt auf vorgegebene Körperzellen beschränken. Durch geeignete "Trafficking" Signalsequenzen kann das gebildete Polypeptid in das Lumen des endoplasmatischen Retikulums dirigiert und in zelluläre Verbreitungswege zur Sekretion oder Expression und Präsentation an der Zelloberfläche gelei- tet werden.For labeling, a DNA can also be applied which has a nucleotide sequence coding for the immunogen. The DNA can contain partial sequences or the entire gene of "keyhole limpet hemocyanine". However, it can also be designed such that it describes only a short peptide, it being possible for several immunogens to be encoded on a single DNA. In the case of naturally occurring genes or sequences derived therefrom, both cDNA and genomic DNA with intron sequences can be applied. In addition to the nucleotides encoding the immunogen, the DNA can contain further sequences which are important for the expression and processing of the immunogen in the livestock organism. These are promoter sequences and sequences which bring about transcription termination and polyadenylation of the mRNA and intracellular "trafficking" of the immunogen. Suitable expression regulators are promoters which ensure a high transcription rate of the DNA, e.g. the "immediate early" promoter of the cytomegalovirus or the promoter of a livestock-specific "house-keeping" gene such as β-actin. However, promoters are also suitable which specifically restrict the expression of the immunogen to predetermined body cells. The polypeptide formed can be directed into the lumen of the endoplasmic reticulum by suitable "trafficking" signal sequences and directed into cellular distribution channels for secretion or expression and presentation on the cell surface.

Der Nachweis der durch die genannten Stoffe induzierten Antikörper erfolgt durch gängige Nachweisverfahren, wie ELISA, EIA und RIA, vorzugsweise unter Verwendung von Teststäbchen, die bsp. in den Tropfsaft des Fleischs des geschlachteten Nutztiers getaucht werden. The antibodies induced by the substances mentioned are detected by common detection methods such as ELISA, EIA and RIA, preferably using test sticks, e.g. be immersed in the dripping juice of the meat of the slaughtered farm animal.

Claims

Patentansprüche claims 1. Verfahren zum Herkunftsnachweis von Nutztieren, insbesondere von Rindern, Schweinen u. dgl . , sowie davon stammen- den Produkten, umfassend die folgenden Schritte:1. Procedure for verifying the origin of farm animals, especially cattle, pigs and the like. the like , as well as products derived from it, comprising the following steps: a) Biologische Markierung des lebenden Nutztiers durch Applikation mindestens eines für das Nutztier und den Menschen unschädlichen Immunogens, unda) biological labeling of the live farm animal by application of at least one immunogen which is harmless to the farm animal and to humans, and b) Nachweis der durch die Applikation des mindestens einen Immunogens gebildeten spezifischen Antikörper oder spezifischen T-Zellen oder deren Rezeptoren im lebenden Nutztier oder in davon stammenden Produkten mittels eines enzymimmuno- logischen oder immunochemisehen Nachweisverfahrens.b) detection of the specific antibodies or specific T cells or their receptors formed by the application of the at least one immunogen in the live farm animal or in products derived therefrom by means of an enzyme-immunological or immunochemical detection method. 2. Verfahren nach Anspruch 1, wobei als Immunogen (e) mindestens ein Protein und/oder Peptid verwendet wird/werden, das/die weder als Impfstoff (e) noch als Arzneimittel, noch über die Nahrungskette, noch über die Umwelt bei der üblichen Nutztierhaltung in den Organismus des Nutztiers gelangt/gelangen und gegen das/die in den Nutztieren natürlicherweise keine Antikörper gebildet werden.2. The method according to claim 1, wherein at least one protein and / or peptide is used as the immunogen (s), which / s neither as a vaccine (s) nor as a medicament, nor via the food chain, nor about the environment in the usual way Livestock farming gets into the organism of the livestock and against which no antibodies are naturally formed in the livestock. 3. Verfahren nach Anspruch 2, wobei ein mit einem Trägerstoff gekoppeltes Protein und/oder Peptid verwendet wird/werden.3. The method according to claim 2, wherein a protein and / or peptide coupled with a carrier is / are used. 4. Verfahren nach einem der vorhergehenden Ansprüche, wobei das Immunogen derart beschaffen ist, daß die gebildeten spezifischen Antikörper dauerhaft im Organismus des Nutztiers verbleiben und mittels enzymimmunologischer oder immunochemi- scher Nachweisverfahren nachweisbar sind. 4. The method according to any one of the preceding claims, wherein the immunogen is such that the specific antibodies formed remain permanently in the organism of the farm animal and can be detected by means of enzyme immunological or immunochemical detection methods. 5. Verfahren nach einem der vorhergehenden Ansprüche, wobei das Immunogen dem Nutztier durch Injektion appliziert wird.5. The method according to any one of the preceding claims, wherein the immunogen is applied to the farm animal by injection. 6. Verfahren nach einem der vorhergehenden Ansprüche, wobei das Immunogen mittels eines Implantats appliziert wird.6. The method according to any one of the preceding claims, wherein the immunogen is applied by means of an implant. 7. Verfahren nach einem der vorhergehenden Ansprüche, wobei das Immunogen über die Schleimhaut, vorzugsweise die Nasen- Schleimhaut, des Nutztiers appliziert wird.7. The method according to any one of the preceding claims, wherein the immunogen is applied via the mucous membrane, preferably the nasal mucous membrane, of the farm animal. 8. Verfahren nach einem der vorhergehenden Ansprüche, wobei eine Mischung von mehreren verschiedenen Immunogenen appliziert wird.8. The method according to any one of the preceding claims, wherein a mixture of several different immunogens is applied. 9. Verfahren nach einem der vorhergehenden Ansprüche, wobei anstelle des Immunogens eine dafür kodierende Desoxyribonu¬ kleinsäure (DNA) appliziert wird.9. The method according to any one of the preceding claims, wherein instead of the immunogen a coding for it Deoxyribonu ¬ small acid (DNA) is applied. 10. Verfahren nach einem der vorhergehenden Ansprüche, wobei der Tropf-/Pressaft des Fleischs des geschlachteten Nutztiers zum Nachweis der spezifischen Antikörper verwendet wird.10. The method according to any one of the preceding claims, wherein the drip / press juice of the meat of the slaughtered farm animal is used to detect the specific antibodies. 11. Verfahren nach einem der vorhergehenden Ansprüche, wobei die Milch oder andere Körpersekrete zum Nachweis der spezifischen Antikörper verwendet wird.11. The method according to any one of the preceding claims, wherein the milk or other body secretions is used to detect the specific antibodies. 12. Verfahren nach einem der vorhergehenden Ansprüche, wobei das Blut, seine Bestandteile oder andere Körperbestandteile des Nutztiers zum Nachweis der spezifischen Antikörper bzw. der spezifischen T-Zellen oder deren Rezeptoren verwendet wird. 12. The method according to any one of the preceding claims, wherein the blood, its components or other body components of the livestock is used to detect the specific antibodies or the specific T cells or their receptors. 13. Verfahren nach einem der vorhergehenden Ansprüche, wobei als enzymimmunologisches oder immunochemisches eines oder mehrere der folgenden Nachweisverfahren verwendet wird/werden: Enzyme Linked Im unosorbent Assay (= ELISA) , Enzyme Im- uno Assay (= EIA) ) , Immunoblot (WESTERN) und Radio Immuno Assay (= RIA) .13. The method according to any one of the preceding claims, wherein one or more of the following detection methods is / are used as enzyme immunological or immunochemical: enzyme linked im unosorbent assay (= ELISA), enzyme immuno assay (= EIA)), immunoblot (WESTERN) and radio immunoassay (= RIA). 14. Verfahren nach einem der vorhergehenden Ansprüche, wobei der enzymimmunologische oder immunochemische Nachweis unter Verwendung von Teststäbchen oder Testchips durchgeführt wird.14. The method according to any one of the preceding claims, wherein the enzyme immunological or immunochemical detection is carried out using test sticks or test chips. 15. Verwendung mindestens eines15. Use at least one a) für Nutztiere und den Menschen unschädlichen Immunogens zur biologischen Markierung des lebenden Nutztiers sowie davon stammender Produkte, und(a) for livestock and harmless immunogen for the biological labeling of live livestock and products derived therefrom, and b) eines enzymimmunologischen oder immunochemisehen Nachweisverfahrens zum Nachweis der durch die Applikation des mindestens einen Immunogens gebildeten spezifischen Antikörper im lebenden Nutztier oder in davon stammenden Produkten.b) an enzyme-immunological or immunochemical detection method for the detection of the specific antibodies formed by the application of the at least one immunogen in the live farm animal or in products derived therefrom. 16. Verwendung nach Anspruch 15, wobei das Immunogen ein Protein und/oder Peptid ist, das weder als Impfstoff noch als16. Use according to claim 15, wherein the immunogen is a protein and / or peptide which is neither a vaccine nor Arzneimittel, noch über die Nahrungskette bei der üblichen Nutztierhaltung in den Organismus des Nutztiers gelangt und gegen das in den Nutztieren natürlicherweise keine Antikörper gebildet werden.Medicines that still enter the organism of the livestock via the food chain in normal livestock farming and against which no antibodies are naturally formed in the livestock. 17. Verwendung nach Anspruch 15 oder 16, wobei das Antigen derart beschaffen ist, daß die gebildeten spezifischen Antikörper dauerhaft im Organismus des Nutztiers verbleiben und mittels enzymimmunologischer oder immunochemischer Nachweisverfahren nachweisbar sind.17. Use according to claim 15 or 16, wherein the antigen is such that the specific antibodies formed remain permanently in the organism of the farm animal and are detectable by means of enzyme immunological or immunochemical detection methods. 18. Verwendung nach Anspruch 16 oder 17, wobei das Protein und/oder Peptid mit einem Trägerstoff gekoppelt ist.18. Use according to claim 16 or 17, wherein the protein and / or peptide is coupled to a carrier. 19. Verwendung nach einem der Ansprüche 15 bis 18, wobei eine Mischung von mehreren verschiedenen Immunogenen verwendet wird.19. Use according to any one of claims 15 to 18, wherein a mixture of several different immunogens is used. 20. Verwendung nach einem der Ansprüche 15 bis 19, wobei anstelle des Immunogens eine dafür kodierende Desoxyribonukleinsäure (DNA) appliziert wird.20. Use according to one of claims 15 to 19, wherein instead of the immunogen a coding for it deoxyribonucleic acid (DNA) is applied. 21. Verwendung nach einem der Ansprüche 15 bis 20, wobei der Tropf-/Pressaft des Fleischs des geschlachteten Nutztiers zum Nachweis der Antikörper verwendet wird.21. Use according to any one of claims 15 to 20, wherein the drip / press juice of the meat of the slaughtered farm animal is used to detect the antibodies. 22. Verwendung nach einem der Ansprüche 15 bis 21, wobei das Blut, seine Bestandteile oder andere Körperbestandteile des22. Use according to any one of claims 15 to 21, wherein the blood, its components or other body components of the Nutztiers zum Nachweis der spezifischen Antikörper bzw. der spezifischen T-Zellen oder deren Rezeptoren verwendet wird.Farm animal is used to detect the specific antibodies or the specific T cells or their receptors. 23. Verwendung nach einem der Ansprüche 15 bis 22, wobei die Milch des Nutztiers oder anderer Körpersekrete zum Nachweis der spezifischen Antikörper verwendet wird.23. Use according to any one of claims 15 to 22, wherein the milk of the farm animal or other body secretions is used to detect the specific antibodies. 24. Verwendung nach einem der Ansprüche 15 bis 23, wobei als enzymimmunologisches oder immunochemisches eines oder mehrere der folgenden Nachweisverfahren verwendet wird/werden: Enzyme Linked Immunosorbent Assay (= ELISA) , Enzyme Immuno Assay (= EIA) , Immunoblot (WESTERN) und Radio Immuno Assay (= RIA) . 24. Use according to any one of claims 15 to 23, wherein one or more of the following detection methods is / are used as enzyme immunological or immunochemical: enzyme linked immunosorbent assay (= ELISA), enzyme immuno assay (= EIA), immunoblot (WESTERN) and radio Immuno Assay (= RIA). 25. Verwendung nach einem der Ansprüche 15 bis 24, wobei zum enzymimmunologischen oder immunochemischen Nachweis Teststäbchen oder Testchips verwendet werden. 25. Use according to any one of claims 15 to 24, wherein test sticks or test chips are used for enzyme immunological or immunochemical detection.
PCT/DE1998/000113 1998-01-14 1998-01-14 Method for detecting the origin of livestock and the products obtained therefrom Ceased WO1999036775A1 (en)

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CA002315023A CA2315023A1 (en) 1998-01-14 1998-01-14 Method for detecting the origin of livestock and products originating therefrom
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WO2004073738A3 (en) * 2003-02-18 2004-10-28 Responsif Gmbh Composition to be administered to a living being and method for marking agents
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WO2001019178A1 (en) 1999-09-15 2001-03-22 november Aktiengesellschaft Gesellschaft für Molekulare Medizin Method for marking an animal and products derived therefrom
DE10011741A1 (en) * 2000-03-13 2001-09-20 Rausch Hans Biologically marking an animal for determining their place of origin comprises the oral administration of an immunogen which produces detectable changes
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DE10130321A1 (en) * 2001-06-22 2003-01-09 Hans-Lorenz Werner Identifying individuals, useful for preventing falsification of urine samples, comprises administering a marker that can be detected in body fluids
WO2004073738A3 (en) * 2003-02-18 2004-10-28 Responsif Gmbh Composition to be administered to a living being and method for marking agents
WO2004078926A3 (en) * 2003-02-28 2004-12-29 Univ Michigan Immunologigal markers

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