KR20230124855A - Portable LAMP-PCR device for POCT - Google Patents
Portable LAMP-PCR device for POCT Download PDFInfo
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- 238000012123 point-of-care testing Methods 0.000 title 1
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- 108020004707 nucleic acids Proteins 0.000 claims abstract description 31
- 238000011901 isothermal amplification Methods 0.000 claims abstract description 27
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- 238000001514 detection method Methods 0.000 claims abstract description 5
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- 241000191967 Staphylococcus aureus Species 0.000 claims abstract description 4
- 229960003085 meticillin Drugs 0.000 claims abstract description 4
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- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 6
- 238000007397 LAMP assay Methods 0.000 claims description 6
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 6
- 230000003321 amplification Effects 0.000 claims description 5
- 238000012408 PCR amplification Methods 0.000 claims description 4
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- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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- B01L7/00—Heating or cooling apparatus; Heat insulating devices
- B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
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- 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
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/689—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for bacteria
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- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
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Abstract
Description
본 발명은 휴대용 등온 핵산 증폭기에 관한 것으로서, 보다 상세하게는 생체물질이나 미생물 등을 검출하여 질병의 유무 또는 바이러스 감염들을 진단하는 데에 있어서, 누구나 현장에서 신속하게 이를 검출할 수 있는 휴대용 핵산 등온증폭기에 관한 것이다. The present invention relates to a portable isothermal nucleic acid amplifier, and more particularly, to a portable nucleic acid isothermal amplifier that anyone can quickly detect in the field in diagnosing the presence or absence of a disease or viral infection by detecting biological materials or microorganisms. It is about.
일반적으로 생체물질이나 미생물 등을 검출함으로써 질병의 유무 또는 바이러스 등의 감염을 진단하기 위해서는 시료를 채취한 후에, 시료에 포함되어 있는 미량의 DNA 및 RNA와 같은 핵산의 유전정보를 분석에 필요한 양 만큼 대량으로 증폭할 필요가 있다. In general, in order to diagnose the presence or absence of a disease or infection such as a virus by detecting a biological material or microorganism, after taking a sample, the genetic information of nucleic acids such as DNA and RNA contained in the sample is collected in an amount necessary for analysis. It needs to be massively amplified.
이를 위해서는 시료 내에 포함되어 있는 핵산을 증폭하기 위하여 다양한 방법을 적용하고 있다. 주로 사용되는 핵산의 증폭 방법으로는, LCR(Ligase Chain Reaction), SDA(Strand Displacement Amplification), NASBA(Nucleic Acid Sequence-Based Amplification), LAMP(Loop-mediated isothermal amplification)와 같은 등온 증폭(isothermal amplification) 방법과, 중합 효소 연쇄 반응(PCR: Polymerase Chain Reaction)과 같은 비등온 증폭(non-isothermal amplification) 방법이 있다. To this end, various methods are applied to amplify the nucleic acid contained in the sample. Mainly used nucleic acid amplification methods include isothermal amplification such as Ligase Chain Reaction (LCR), Strand Displacement Amplification (SDA), Nucleic Acid Sequence-Based Amplification (NASBA), and Loop-mediated isothermal amplification (LAMP). method, and a non-isothermal amplification method such as polymerase chain reaction (PCR).
비등온 증폭 방법의 대표적인 예인 중합 효소 연쇄 반응은 열변성(Denaturation), 결합(Annealing), 및 신장(Extension) 등 일련의 온도 효소 반응 단계로 진행되고, 이들 반응 단계는 각각 일정한 온도 범위에서 진행되어야 양질의 핵산을 고수율로 수득할 수 있다. Polymerase chain reaction, which is a typical example of a non-isothermal amplification method, proceeds through a series of temperature enzyme reaction steps such as thermal denaturation, annealing, and extension, and each of these reaction steps must be carried out at a certain temperature range. High-quality nucleic acids can be obtained in high yield.
등온증폭 방법 및 비등온 증폭방법 모두 반응 온도를 일정하게 유지하는 것이 바람직한 결과를 수득할 수 있는 중요한 인자이다. For both the isothermal amplification method and the non-isothermal amplification method, keeping the reaction temperature constant is an important factor for obtaining desirable results.
위와 같이 반응 온도를 일정하게 유지하기 위하여 종래의 휴대용 핵산 증폭기의 경우에는 휴대용 증폭기 내에 축전기 등에 의해 전원을 공급하고, 전원에 의해서 발열하는 광학소자 등을 구비하여 상기 휴대용 증폭기의 온도를 유지하는 방식을 적용하고 있다. In order to keep the reaction temperature constant as described above, in the case of a conventional portable nucleic acid amplifier, power is supplied by a capacitor in the portable amplifier, and an optical element that generates heat by the power supply is provided to maintain the temperature of the portable amplifier. are applying
본 발명은 상기와 같은 전원에 의해서 발열하는 광학소자를 사용하지 않고서도 반응온도를 유지할 수 있는 휴대용 등온증폭기를 제공하기 위한 것이다. An object of the present invention is to provide a portable isothermal amplifier capable of maintaining a reaction temperature without using an optical element that generates heat by the power source as described above.
본 발명은 생체물질 또는 미생물을 검출함으로써 질병의 진단 및 바이러스 등의 감염을 현장에서 신속하고 정확하게 판단하기 위한 휴대용 등온증폭 핵산 증폭기에 있어서, 상기 증폭기의 반응온도를 일정하게 유지시킬 수 있는 전원 공급장치와 광학소자 등을 사용하지 않고, 휴대용 발열수단을 이용해서 상기 증폭기를 가동시킬 수 있는 휴대용 핵산 등온증폭기를 제공하고자 하는 것이다. The present invention is a portable isothermal amplifying nucleic acid amplifier for quickly and accurately determining disease diagnosis and virus infection in the field by detecting biomaterials or microorganisms, and a power supply device capable of maintaining a constant reaction temperature of the amplifier. It is an object of the present invention to provide a portable nucleic acid isothermal amplifier capable of operating the amplifier using a portable heating means without using an optical device or the like.
본 발명은 상기와 같은 과제를 해결하기 위하여, 본 발명에서 제공하는 증폭기의 상부에는 시료를 채취한 다수의 PCR 튜브를 고정하는 튜브홀더와 뚜껑을 구비하고, 상기 증폭기의 하부에는 손난로 등의 발열수단을 삽입할 수 있는 수납공간을 설치함으로써 본 발명을 완성하게 되었다. In order to solve the above problems, the present invention provides a tube holder and a lid for fixing a plurality of PCR tubes from which samples are taken at the upper part of the amplifier provided by the present invention, and a heating device such as a hand warmer is provided at the lower part of the amplifier. The present invention has been completed by installing a storage space into which means can be inserted.
본 발명에서 제공하는 휴대용 핵산의 등온증폭기는 현장에서 채취한 샘플에 포함되어 있는 핵산 등을 신속하게 현장에서 증폭할 수 있는 기기로서 누구나 간편하게 사용할 수 있고, 또한 반응온도를 유지하기 위하여 종래에 사용되어왔던 전원의 공급에 의한 발열수단을 사용하지 않고, 접근성이 용이한 손난로 등의 발열수단을 이용하여 핵산의 증폭을 위한 온도를 유지할 수 있으므로 사용이 간편할 뿐만 아니라, 제작이 용이하고 제작비용도 저렴하다는 장점이 있다. The portable nucleic acid isothermal amplifier provided by the present invention is a device that can quickly amplify nucleic acid contained in a sample taken in the field on the spot, and can be used conveniently by anyone, and is conventionally used to maintain the reaction temperature. Since the temperature for nucleic acid amplification can be maintained using an easily accessible heating means such as a hand warmer without using a heating means by the supply of power, it is not only easy to use, but also easy to manufacture and reduce the manufacturing cost. It has the advantage of being cheap.
도 1은 본 발명에서 제공하는 휴대용 등온증폭장치를 나타낸다.
도 2는 본 발명에서 제공하는 휴대용 등온증폭장치의 수납공간을 설명한 것이다.
도 3은 본 발명에서 제공하는 휴대용 등온증폭장치를 이용하여 MRSA를 검출하는 실시예의 결과를 나타낸다.1 shows a portable isothermal amplifier provided by the present invention.
2 illustrates the storage space of the portable isothermal amplifier provided by the present invention.
Figure 3 shows the results of an embodiment of detecting MRSA using a portable isothermal amplifier provided by the present invention.
이하 첨부된 도면을 참조로 본 발명의 바람직한 실시예를 상세히 설명하기로 한다. 이에 앞서, 본 명세서 및 청구범위에 사용된 용어나 단어는 통상적이거나 사전적인 의미로 한정해서 해석되어서는 아니되며, 발명자는 그 자신의 발명을 가장 최선의 방법으로 설명하기 위해 용어의 개념을 적절하게 정의할 수 있다는 원칙에 입각하여 본 발명의 기술적 사상에 부합하는 의미와 개념으로 해석되어야만 한다. 따라서, 본 명세서에 기재된 실시예와 도면에 도시된 구성은 본 발명의 가장 바람직한 일 실시예에 불과할 뿐이고 본 발명의 기술적 사상을 모두 대변하는 것은 아니므로, 본 출원시점에 있어서 이들을 대체할 수 있는 다양한 균등물과 변형예들이 있을 수 있음을 이해하여야 한다. Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings. Prior to this, the terms or words used in this specification and claims should not be construed as being limited to the usual or dictionary meaning, and the inventor appropriately uses the concept of the term in order to explain his/her invention in the best way. It should be interpreted as a meaning and concept consistent with the technical idea of the present invention based on the principle that it can be defined. Therefore, since the embodiments described in this specification and the configurations shown in the drawings are only one of the most preferred embodiments of the present invention and do not represent all of the technical ideas of the present invention, various alternatives may be used at the time of this application. It should be understood that there may be equivalents and variations.
병원이나 진료소 등의 현장에서 미생물 또는 바이러스의 감염을 진단하기 위해서는 상기 튜브를 이용하여 현장에서 시료를 수득한 후에 이를 상기 미생물 또는 바이러스를 검출하는 실험실 등에 이송한 후에, 상기 실험실 등에 비치되어 있는 PCR 등온증폭기 등을 이용하여 튜브에 포함되어 있는 핵산을 증폭하고, 이를 형광진단 등의 검출방법을 통해 진단하여야 한다. In order to diagnose microbial or virus infection in a hospital or clinic, a sample is obtained in the field using the tube, transferred to a laboratory that detects the microbial or virus, and PCR isotherms provided in the laboratory. The nucleic acid contained in the tube should be amplified using an amplifier, etc., and diagnosed through a detection method such as fluorescence diagnosis.
따라서 상기와 같은 과정 중에 현장에서 채취한 시료가 오염되거나 형질이 변환된 가능성이 있어서, 이와 같은 문제점을 해결하기 위해서는 현장에서 미생물 또는 바이러스를 신속하게 검출하는 것이 중요한 사항이다. Therefore, there is a possibility that the sample collected in the field may be contaminated or transformed during the above process, and in order to solve this problem, it is important to quickly detect microorganisms or viruses in the field.
상기와 같은 문제점을 해결하기 위하여 본 발명에서 제공하는 휴대용 등온증폭장치(100)는 현장에서 시료에 포함되어 있는 타겟 핵산을 증폭할 수 있는 PCR 튜브를 사용하고, 상기 증폭과정 중에 핵산이 효율적으로 증폭될 수 있도록 반응 온도를 일정하게 유지할 수 있는 발열수단을 상기 휴대용 등온증폭장치의 내부에 구비하는 것을 특징으로 한다. In order to solve the above problems, the portable isothermal amplification device 100 provided by the present invention uses a PCR tube capable of amplifying the target nucleic acid contained in the sample in the field, and the nucleic acid is efficiently amplified during the amplification process. It is characterized in that a heating means capable of maintaining a constant reaction temperature is provided inside the portable isothermal amplifier.
본 발명에서 제공하는 휴대용 등온증폭장치는 병원이나 보건소 등의 현장에서 미생물 및 바이러스 등을 신속하게 검출할 수 있는 휴대용 등온증폭장치에 대한 것으로서, 상기 휴대용 등온증폭장치(100)는 PCR법을 이용하여 검출대상이 되는 핵산을 증폭할 때에 고리매게등온증폭방법(LAMP)을 적용하는 것을 특징으로 하고 있다. The portable isothermal amplification device provided by the present invention relates to a portable isothermal amplification device that can quickly detect microorganisms and viruses in a hospital or public health center. The portable isothermal amplification device 100 uses the PCR method It is characterized by applying a cyclic isothermal amplification method (LAMP) when amplifying a nucleic acid to be detected.
본 발명의 휴대용 등온증폭장치는 상부에는 시료가 포함되어 있는 PCR 튜브(10)를 고정할 수 있는 PCR 튜브 삽입부(20)가 형성되어 있고, 상기 PCR 튜브(10)를 삽입한 후에 상기 튜브를 고정하기 위한 커버(30)가 형성되어 있다. The portable isothermal amplification device of the present invention has a PCR tube insert 20 capable of fixing a PCR tube 10 containing a sample at the top, and after inserting the PCR tube 10, the tube A cover 30 for fixing is formed.
또한, 본 발명에서 제공하는 휴대용 등온증폭장치(100)의 하부에는 상기 PCR 튜브(10)내에 포함되어 있는 핵산을 LAMP법에 의해서 증폭하기 위한 가장 중요한 조건인 반응온도를 일정하게 유지하기 위한 손난로 등과 같은 발열수단(40)을 수납하기 위한 수납공간(50)이 형성되어 있고, 상기 수납공간(50)을 외부와 차단하여 상기 발열수단(40)에 의해서 발생되는 열을 본 발명의 휴대용 등온증폭기(100)의 내부에서만 유지되도록 하는 수납공간 커버(60)가 구비되어 있다. In addition, at the bottom of the portable isothermal amplification device 100 provided by the present invention, a hand warmer is provided to keep the reaction temperature constant, which is the most important condition for amplifying the nucleic acid contained in the PCR tube 10 by the LAMP method. A storage space 50 for accommodating a heating means 40 such as a lamp is formed, and the storage space 50 is blocked from the outside to transfer heat generated by the heating means 40 to the portable isothermal amplifier of the present invention. A storage space cover 60 is provided so as to be maintained only inside the 100.
본 발명에서 제공하는 휴대용 등온증폭장치(100)의 내부에 포함되는 상기 PCR 튜브는 시료에 포함되어 있는 DNA 또는 RNA 등의 타겟 핵산을 증폭하기 위한 PCR 증폭시약과 더불어 상기 타겟 핵산을 검출하기 위한 프로브와 형광물질로 이루어지는 진단시약이 모두 포함되어 있다. The PCR tube included in the portable isothermal amplification device 100 provided by the present invention includes a PCR amplification reagent for amplifying a target nucleic acid such as DNA or RNA contained in a sample and a probe for detecting the target nucleic acid. and diagnostic reagents composed of fluorescent substances are all included.
따라서, 현장에서 시료를 채취한 후에 상기 PCR 튜브(10)에 시료를 투입하고, 이를 본 발명에서 제공하는 휴대용 등온증폭장치(100)에 삽입하고, 일정한 시간이 지나면, 상기 발열수단에 의해서 상기 휴대용 등온증폭장치에 일정한 온도가 유지되고, 이로 인해 PCR 튜브 내에 포함되어 있는 핵산이 증폭하고, 튜브 내에 있는 프로브와 반응하여 튜브 내의 시약의 색이 변하게 된다. Therefore, after taking a sample in the field, the sample is put into the PCR tube 10, inserted into the portable isothermal amplification device 100 provided by the present invention, and after a certain period of time, the portable isotherm is activated by the heating means. A constant temperature is maintained in the isothermal amplification device, whereby the nucleic acid contained in the PCR tube is amplified and reacts with the probe in the tube to change the color of the reagent in the tube.
이와 같은 색의 변화를 이용하여 시료에 포함되어 있는 특정한 미생물 또는 바이러스의 존재를 현장에서 신속하게 확인할 수 있고, 또한 미생물이나 바이러스에 의한 현장의 감염여부도 확인할 수 있다. By using such a color change, the presence of a specific microorganism or virus contained in the sample can be quickly confirmed in the field, and also whether or not the microorganism or virus is infected in the field can be confirmed.
본 발명에서 제공하는 휴대용 등온증폭장치(100)의 내부에 거치되는 PCR 튜브(10)의 내부에 투입되는 시약은 어떠한 종류의 타겟 핵산이든 이를 증폭할 수 있는 PCR 증폭 시약과 상기 타겟 핵산을 검출할 수 있는 진단시약을 포함하여 이루어진다. The reagents introduced into the PCR tube 10 mounted inside the portable isothermal amplification device 100 provided by the present invention are PCR amplification reagents capable of amplifying any kind of target nucleic acid and those capable of detecting the target nucleic acid. It is made up of possible diagnostic reagents.
따라서 본 발명에서 제공하는 휴대용 등온증폭기는 상기 PCR 튜브(10)의 내부에 투입되는 시약에 따라서 현장에서 감염이 의심되는 어떠한 종류의 미생물 또는 바이러스에 대해서 적용이 가능하다. Therefore, the portable isothermal amplifier provided in the present invention can be applied to any kind of microorganism or virus suspected of being infected in the field according to the reagent injected into the PCR tube 10.
본 발명에서 제공하는 휴대용 등온증폭장치(100)에서 상기 PCR 튜브내에 투입되는 시료의 핵산의 PCR 증폭을 위하여 열을 제공하는 발열수단(40)은 손난로, 발열팩, 발열패드 등을 본 발명의 휴대용 등온증폭장치(100)에 구비되어 있는 수납공간(50)에 직접 끼워 넣을 수 있다. In the portable isothermal amplification device 100 provided in the present invention, the heating means 40 that provides heat for PCR amplification of the nucleic acid of the sample put into the PCR tube is a hand warmer, a heating pack, a heating pad, etc. It can be directly inserted into the storage space 50 provided in the portable isothermal amplifier 100.
또한, 상기 발열수단으로서는 철의 산화를 통하여 발생하는 열을 이용할 수 있는 발열 조성물을 상기 수납공간(50)에 직접 넣어 사용할 수 있는데, 상기 발열 조성물은 철가루, 소금, 활성탄, 톱밥, 질석, 규조토 및 물 등을 포함하여 이루어지는 고체형 조성물일 수 있다. In addition, as the heating means, a heating composition capable of using heat generated through oxidation of iron may be directly put into the storage space 50 and used. The heating composition includes iron powder, salt, activated carbon, sawdust, vermiculite, and diatomaceous earth. And it may be a solid composition comprising water and the like.
상기 발열 조성물은 철가루가 산소와 물을 만나면 발생하는 철의 산화과정에서 발생하는 열을 이용한다. The exothermic composition uses heat generated in the oxidation process of iron generated when iron powder meets oxygen and water.
또한 상기 발열 조성물로서는 액체형 발열 조성물을 사용할 수 있는데, 상기 액체형 조성물은 티오황산나트륨 또는 아세트산나트륨과 금속압전체를 포함하여 이루어진다. In addition, a liquid type exothermic composition may be used as the exothermic composition, and the liquid composition includes sodium thiosulfate or sodium acetate and a metal piezoelectric body.
본 발명에서 사용되는 발열 수단은 상기의 발열 조성물 이외에도 수납이 가능하면 어떠한 발열수단이라도 모두 사용될 수 있다. As the heat generating means used in the present invention, any heat generating means other than the above heat generating composition can be used as long as it can be accommodated.
도 2는 본 발명에서 제공하는 휴대용 등온증폭장치의 수납공간은 자세히 설명한 것이다. 2 is a detailed description of the storage space of the portable isothermal amplifier provided by the present invention.
도 2의 상부 도면은 수납공간의 평면도를 나타내는데, 수납공간의 내부에는 상기 발열수단의 온도를 측정할 수 있는 열감지스티커(70)가 구비되어 있다. 상기 열감지스티커(70)는 수납공간의 상부에서 발열수단에 접촉하도록 설치되어 있고, 상기 열감지스티커(70)에 의해서 발열수단에서 발생하는 열의 온도를 측정할 수 있는데, 발열수단에서 열이 발생할 경우에 색이 변하면서 특정한 온도(약 60℃)를 상회할 경우에는 다른 색으로 완전히 변화한다. The upper drawing of FIG. 2 shows a plan view of the storage space, and a heat sensing sticker 70 capable of measuring the temperature of the heating means is provided inside the storage space. The heat detection sticker 70 is installed to contact the heat generating means at the top of the storage space, and the temperature of the heat generated from the heat generating means can be measured by the heat detection sticker 70, where heat is generated from the heat generating means. When the temperature exceeds a certain temperature (about 60 ℃) while the color changes, it completely changes to a different color.
이와 같은 온도의 변화를 육안으로 관찰할 수 있고, 예를 들면 흰색을 나타내던 상기 열감지스티커(70)가 발열수단에서 발행하는 열에 의해서 적색을 나타낼 때에 핵산의 등온증폭이 활발하게 일어나고 있음을 알 수 있다. Such a change in temperature can be observed with the naked eye, and it can be seen that, for example, isothermal amplification of nucleic acids is actively occurring when the heat-sensing sticker 70, which was white, turns red due to the heat generated from the heating means. can
도 2의 하부 도면은 수납공간을 분리한 것인데, 수납공간의 내부의 하부에는 단열재(80)가 구비되어 있어서, 발열수단에서 발생하는 열이 외부로 이탈되는 것을 방지하여 핵산의 효율적인 등온증폭이 수행되도록 한다. The lower drawing of FIG. 2 shows the separated storage space. Insulator 80 is provided at the lower part of the storage space to prevent the heat generated from the heating unit from escaping to the outside, so that efficient isothermal amplification of nucleic acids is performed. Let it be.
<실시예><Example>
다음은 본 발명에 의해 제공되는 휴대용 등온증폭장치를 이용하여 메티실린 내성 황색포도상구균(MRSA)을 검출한 실시예에 대해서 설명한다.Next, an embodiment in which methicillin-resistant Staphylococcus aureus (MRSA) was detected using the portable isothermal amplifier provided by the present invention will be described.
본 발명에서 제공하는 휴대용 등온증폭장치를 이용하여 MRSA를 검출하기 위하여, 상기 PCR 튜브(10)에는 MRSA의 농도를 변화시킨 시약을 25㎕를 주입하였고, 상기 발열 수단으로서는 철가루, 활성탄, 염화나트륨, 질석 및 고흡습성 폴리머로 이루어지는 고체형 발열 수단은 상기 수납공간(50)에 넣고 반응시킨 후에 반응시간이 25분이 되는 시간부터 5분 간격으로 상기 PCR 튜브에 주입된 시약의 색의 변화를 관찰하였다.In order to detect MRSA using the portable isothermal amplification device provided by the present invention, 25 μl of a reagent having a changed concentration of MRSA was injected into the PCR tube 10, and iron powder, activated carbon, sodium chloride, The solid-type heating means made of vermiculite and highly hygroscopic polymer was placed in the storage space 50 and reacted, and then the color change of the reagent injected into the PCR tube was observed at 5-minute intervals from the time when the reaction time was 25 minutes.
도 3에 제시된 바와 같이, 본 발명에서 제공하는 휴대용 등온증폭장치를 이용하여 MRSA를 검출할 경우에 MRSA의 농도가 극소량이라 하더라도 신속하게 검출할 수 있었다. As shown in FIG. 3, when MRSA was detected using the portable isothermal amplification device provided in the present invention, even if the concentration of MRSA was extremely small, it could be quickly detected.
도 3에서 각 숫자는 PCR 튜브에 포함되어 있는 MRSA의 농도를 나타내고, PC는 양성 컨트롤, NC는 음성 컨트롤을 나타내고, 도 3a는 상기 PCR 튜브를 휴대용 등온증폭장치(100)에 삽입하기 전의 상태를 나타내고, 도 3b는 상기 PCR 튜브를 휴대용 등온증폭장치(100)에 삽입한 후의 시간 및 MRSA의 농도에 따른 시약의 색변화를 나타낸다. In Figure 3, each number represents the concentration of MRSA contained in the PCR tube, PC represents a positive control, NC represents a negative control, and Figure 3a shows the state before inserting the PCR tube into the portable isothermal amplifier 100. 3b shows the color change of the reagent according to the time and MRSA concentration after the PCR tube is inserted into the portable isothermal amplification device 100.
본원 발명을 상기의 실시예 및 도면에 의해서 설명하였으나, 본원 발명의 기술적 사상은 상기 실시예 등에 의해 한정되지 않고, 본원 발명의 기술적 특징을 포함하는 모든 구성까지 확장될 수 있다는 것은 통상의 기술자에게는 주지의 사항이다. Although the present invention has been described with the above embodiments and drawings, it is well known to those skilled in the art that the technical spirit of the present invention is not limited by the above embodiments and can be extended to all configurations including the technical features of the present invention. is the matter of
본 발명은 산업상 이용가능하다.The present invention is industrially applicable.
100 휴대용 등온증폭장치 10 PCR 튜브
20 튜브 삽입부 30 커버
40 발열수단 50 수납공간
60 수납공간 커버
70 열감지스티커 80 단열재100 portable isothermal amplifier 10 PCR tubes
20 Tube insert 30 Cover
40 heat source 50 storage space
60 Storage Cover
70 Thermal sticker 80 Insulator
Claims (1)
상기 수납공간(50)의 내부에는 외부와 열의 출입을 차단하는 단열재(80)와 색변화에 의해 상기 발열수단(40)의 온도를 감지하여 핵산의 등온증폭이 발생하는 것을 가시적으로 확인할 수 있는 열감지스티커(70)가 구비되어 있고,
상기 PCR 튜브(10)의 내부에 주입되는 시약은 메티실린 내성 황색포도상구균의 타겟 핵산을 증폭하기 위한 PCR 증폭시약과 상기 타겟 핵산을 검출하기 위한 진단시약을 모두 포함하여 형성되며,
상기 발열수단(40)은 철가루, 소금, 활성탄, 톱밥, 질석 및 물을 포함하여 형성되는 고체형 발열 조성물인 것을 특징으로 하는 휴대용 등온증폭장치
It relates to a portable isothermal amplification device (100) for amplification of nucleic acid extracted from methicillin-resistant Staphylococcus aureus using loop-mediated isothermal amplification (LAMP), wherein a PCR tube ( 10) is formed, a cover 30 covering the tube insertion part 20 is provided, and a power supply device is provided at the bottom of the portable isothermal amplifier 100. A storage space 50 capable of accommodating a heating means 40 that does not require a storage space 50 and a storage space cover 60 that blocks the storage space from the outside are provided,
Inside the storage space 50, heat insulator 80 that blocks heat from entering and leaving the outside and heat that can be visually confirmed that isothermal amplification of nucleic acid occurs by detecting the temperature of the heating means 40 by color change A detection sticker 70 is provided,
The reagent injected into the PCR tube 10 includes both a PCR amplification reagent for amplifying the target nucleic acid of methicillin-resistant Staphylococcus aureus and a diagnostic reagent for detecting the target nucleic acid,
The heating means 40 is a portable isothermal amplification device, characterized in that the solid heating composition formed by including iron powder, salt, activated carbon, sawdust, vermiculite and water
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| KR101033554B1 (en) * | 2010-08-31 | 2011-05-11 | 두산동아 주식회사 | Thermosensing label having temperature indicating function |
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| KR20180055610A (en) * | 2016-11-17 | 2018-05-25 | 변창훈 | Cool Hand fire in the form of gloves |
| KR20180117454A (en) | 2017-04-19 | 2018-10-29 | 주식회사 넥서스비 | Portable real-time pcr measuring instrument |
| KR102113957B1 (en) * | 2019-12-05 | 2020-05-21 | 제주특별자치도(제주특별자치도해양수산연구원장) | Portable Gene Extraction and Analysis Device for PCR |
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- 2021-02-26 KR KR1020210026615A patent/KR20220122244A/en active Application Filing
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| KR200430299Y1 (en) * | 2006-08-01 | 2006-11-13 | (주)아모레퍼시픽 | A pack for eye warming massage |
| KR20100119127A (en) * | 2009-04-30 | 2010-11-09 | 단국대학교 산학협력단 | Chip for cell amplification reaction and isothermal amplification device using the same |
| KR101033554B1 (en) * | 2010-08-31 | 2011-05-11 | 두산동아 주식회사 | Thermosensing label having temperature indicating function |
| WO2015021425A1 (en) * | 2013-08-09 | 2015-02-12 | The Regents Of The University Of California | Digital fluid sample separation apparatus and methods for one-step quantitative sample analysis |
| KR20180055610A (en) * | 2016-11-17 | 2018-05-25 | 변창훈 | Cool Hand fire in the form of gloves |
| KR20180117454A (en) | 2017-04-19 | 2018-10-29 | 주식회사 넥서스비 | Portable real-time pcr measuring instrument |
| KR102113957B1 (en) * | 2019-12-05 | 2020-05-21 | 제주특별자치도(제주특별자치도해양수산연구원장) | Portable Gene Extraction and Analysis Device for PCR |
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| KR102628993B1 (en) | 2024-01-25 |
| KR20220122244A (en) | 2022-09-02 |
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