TW570982B - Nucleic acid as marker for product anti-counterfeiting and identification - Google Patents

Abstract

This invention features a method of labeling objects for anti-counterfeit purpose, especially refers to a method employing medium containing ribonucleic acid for product anti-counterfeit labeling and authenticity verification by PCR method. The procedure involves label objects with medium which can readily mix and protect ribonucleic acid. For verification of authenticity, the medium is removed and extracted for ribonucleic acid, which is then amplified by PCR method for comparison.

Description

570982 _Case No. 89108443_ Modification of the month and month _ V. Description of the invention (1) The present invention is a method for marking articles to achieve anti-counterfeiting purposes, especially a method using RNA as a product anti-counterfeiting mark and verification method. The medium mixed with RNA is labeled on the target, remove the medium during the test, extract the RNA with the solvent and recover, and then use the polymerase chain reaction to amplify the RNA signal for inspection. According to the results, identify the target Authenticity. [Background note]

One of the problems that often occurs in product manufacturing and marketing is the problem of counterfeiting and counterfeiting. Counterfeit and fake products use the appearance and brand of the genuine product to use the image of the genuine product for the purpose of profit; because counterfeits and fake products are usually similar in appearance and brand, but are made of inferior materials; there are also sales of genuine quality Counterfeit products, but because it does not cost advertising and marketing costs, it can capture the market for genuine products at lower prices. In addition, items with high unit prices, such as famous paintings, jewelry, and souvenirs, as well as items with financial functions and values, such as credit cards, passbooks, and valuable securities, often face the problem of counterfeiting. Such problems will not only damage the goodwill of genuine products and affect the sales of genuine products, but also hinder the financial order and ability of invention and innovation of a country. necessity. Existing methods to prevent counterfeiting, in addition to winning consumers' trust with their obviously special shape and quality, also have other ways to achieve anti-counterfeiting purposes, such as magnetic barcodes on passbooks and laser label patterns on financial cards. And secrets displayed only at special wavelengths, there are also proteins, antibodies, fluorescent substances, etc. added to protected objects, which are inspected in special ways to verify their integrity

Page 6 570982 _Case No. 89108443_Year Month Amendment_ V. Description of the Invention (2); Such means are intended to create a technical or methodic obstacle that makes it difficult for imitation and counterfeiters to imitate easily. What is provided is the protection of the technical barrier, but such protection can still be easily imitated by those with the same technical ability. Therefore, the present invention provides a more specific anti-counterfeiting method that cannot be counterfeited by those with the same technical ability. [Brief description of the invention]

The invention utilizes the characteristic of sequence specificity of ribonucle 1 eicacid. After the ribonucleic acid is mixed with the medium, the medium with the ribonucleic acid can be coated on the target to be forged or penetrated into the target. Inside the object, the RNA carried on the target object can be checked to determine whether the target object was originally labeled.

The characteristics of the medium are that it can be fully mixed with RNA and is not part of the target. The composition of the RNA is designed to have a specific length and sequence, and this sequence can only use a specific polymerase during detection. Chain reaction primers can detect and verify the authenticity of the RNA contained in the medium. During the operation, the medium is first dissolved in a solvent to liquefy the medium, and then a quantitative and known sequence of ribonucleic acid is added to the medium solution and uniformly mixed. At this time, the medium solution with the ribonucleic acid can be used to coat or fill the target; After evaporation, the target is a medium with mixed RNA. Once you want to verify whether the object is a target with a specific RNA sequence, you can dissolve a small part of the medium on the target in the solvent and add Another solvent that has high RNA solubility but does not affect the target is fully mixed. At this time, most of the RNA will be dissolved in the high RNA solubility solvent, and then the different solvents are separated and separated by centrifugation. RNA extracted at this time

Page 7 570982 _Case No. 89108443_Year Month Amendment _ V. Description of the invention (3) The concentration is sufficient to verify the authenticity of the RNA with specific primers from the polymerase chain reaction; by this, if the target If there is the original RNA, the polymerase chain reaction will amplify the original RNA into millions of times the same size of the RNA, and if the test object does not contain the original RNA, the polymerase chain reaction The reaction cannot carry out the amplification reaction of ribonucleic acid, so by comparing the size and quantity of the products, it can be discriminated whether the target object to be identified is the one originally marked.

Because the RNA sequence is specific, the correct nucleic acid size can only be obtained by using specific primers when performing polymerase chain reaction verification, and because the concentration of RNA in the medium is extremely low, it cannot be obtained by gene transfer. The sequence and size of the nucleic acid is known, so the confidentiality and specificity are extremely high. [Detailed description of the invention] The present invention utilizes the sequence characteristics of RNA. The principle that it cannot be copied unless the content at both ends of the sequence is known. It saves the RNA in the medium, and then marks the medium on the target as an identity mark. . When you want to verify the authenticity of the target at a certain time, you only need to remove the medium to test the RNA contained in it to determine the authenticity of the target.

The so-called ribonucleic acid is a general term for deoxyribonucleic acid and ribonucleic acid. The source of ribonucleic acid can come from organic organisms such as animals, plants, bacteria, viruses, etc. But it can also artificially synthesize the ribonucleic acid of vector or fragment. The characteristic of ribonucleic acid is that it The special sequence can be copied by polymerase chain reaction using corresponding primers, but the prerequisite is that the sequences at both ends of the nucleic acid fragment to be copied must be known in order to design appropriate primers for replication.

Page 8 570982 Case No. 89108443 _η Amendment 5. Explanation of invention (4) System reaction. The so-called medium is a homogeneous mixture of the intermediaries of other objects, and must be able to cover the plasticity and appropriate strength. For example: polycarbonate, the so-called target is solid; liquids such as lubricating day, jewelry, and financial products can be The method of marking the target object can be a credit card. It can be mixed in an object such as a seal. Examples of inventions are used to contain RNA and used to attach or mix substances. A good medium must be able to protect the RNA from the RNA. When damaged, the medium must also be able to easily attach to the marked object, such as acrylic or plastic. Articles marked as anti-counterfeiting can be liquid or oil, gasoline and other oil products; solid objects such as antiques, credit cards, etc. have commemorative or practical value. 0 is to apply the medium on the surface of the target, such as inside an object such as Ink and oil can be filled during the day and can be designed according to actual needs. The following is the first embodiment of the present invention. Example 1: Polycarbonate is used as a medium to label 8 0 b ρ DNA on a plastic film. Material: Polycarbonate 95% alcohol propane gas imitation DuPont polycarbonate Chinese Pharmacopoeia Taiwan Merck UN 1 0 9 0 Merck Taiwan UN1 888 The 8 0 0 b ρ synthesized by polymerase chain reaction alcohol

Page 9 570982 Case No. 89108443 Amendment V. Description of the invention (5) Add acetone to dissolve it, and then mix it thoroughly with aerosol-soluble polycarbonate and apply it on the plastic film. After drying, place them in a 40 ° C refrigerator, in a dark place, or expose them to the sun for one day before recycling. When recovering, cut a small number of membranes, dissolve them in aerosol, and mix them with buffer solution and centrifuge to obtain a clear solution. Take a small amount of clarification solution as a template for polymerase chain reaction. The polymerase chain reaction products were separated by electrophoresis and stained. Please refer to the first figure, which shows an example using polycarbonate as a medium and 800 b p DNA as a label. From left to right, L1 is a 100bp DNA standard, L2 is a dark treatment, L3, L4, and L5 are exposed to the sun, and L6, L7, and L8 are a 40 C refrigerator. The results showed that all the treatments were as expected, with a smooth recovery of 8 0 0 b p DNA. Example 2: Labeling a human DNA genome on a plastic film using polycarbonate as a medium. Material: Polycarbonate S, 95% Alcohol, Acetone, Chloroform Taiwan> 'Bend Du Pont Polycarbonate Chinese Pharmacopoeia Taiwan Merck UN 1 0 9 0 Taiwan Merck UN 1 8 8 8 DNA from human white blood cells extracted with DNA Acetone is dissolved, and then fully mixed with polycarbonate dissolved in chloroform, and then applied to a plastic film. After drying, place in a 40 ° C refrigerator, a dark place, or expose to sunlight for one day before recycling. Cut off a small number of membranes during recovery and dissolve in chloroform

Page 10 570982 _ case number 89108443 _ month and month __ V. Description of the invention (6) Then, the buffer solution is thoroughly mixed and centrifuged to obtain a clear solution. A small amount of clear solution was used as a template for polymerase chain reaction. The polymerase chain reaction product is separated by electrophoresis and stained. Please refer to the second figure, which shows a conventional example using the polycarbonate as a medium ' From left to right, L1 is a 100 bp DNA standard, L2 and L3 are the results of 1 ul clarified solution as a template, L 4 and L 5 are the results of 2 u 1 clarified solution as a template, and L 6 is the result of no template added. Negative control group, L 7 is the positive control group of human genome DNA. The results showed that all treatments, except L 3 were less pronounced, recovered 600 b p human genes as expected.

Page 11 570982 _Case No. 89108443_Year Month Amendment _ Brief Description of the Drawings The first diagram shows that polycarbonate is used as the medium, and 800 bp DNA is labeled on a plastic film. After recycling, it is polymerized. Example in which the signal is amplified by an enzyme chain reaction and separated by electrophoresis to obtain 800 bp DNA. The second picture shows the use of polycarbonate as a medium to label the human DNA genome on a plastic membrane. After recovery, the signal is amplified by polymerase chain reaction and separated by electrophoresis to obtain 600 bp deoxyribose. Example of Nucleic Acid

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Claims (1)

570982 _Case No. 89108443_Year_Month__ VI. Application for Patent Scope Agent. 7. The method of applying the RNA as an anti-counterfeiting mark on a product as described in item 6 of the scope of the patent application, wherein the aforementioned organic solvent may be alcohol, acetone or chloroform. 8. As the method of anti-counterfeiting labeling of RNA as described in item 1 of the scope of patent application, the aforementioned target objects can be inks, lubricants, gasolines, antiques, famous items, jewelry, financial credit cards, or seals. Page 14