PORTABLE DEVICE FOR LAMP PCR

Abstract

Provided is a portable isothermal amplification device for amplifying a nucleic acid using an isothermal amplification method, the device comprising: a tube insertion part into which a PCR tube is to be inserted, which is provided in an upper portion of the portable isothermal amplification device; a cover for covering the tube insertion part; a storage space for accommodating a heating device and a storage space cover for blocking the storage space from an outside, which is provided in a lower portion of the portable isothermal amplification device; and a heat insulator for blocking heat from the outside and a heat-sensing sticker capable of measuring a temperature of heat generated by the heating device, which is provided inside the storage space.

Claims

1. A portable isothermal amplification device for amplifying a nucleic acid using an isothermal amplification method, the device comprising: a tube insertion part into which a PCR tube is to be inserted, which is provided in an upper portion of the portable isothermal amplification device; a cover for covering the tube insertion part; a storage space for accommodating a heating device and a storage space cover for blocking the storage space from an outside, which is provided in a lower portion of the portable isothermal amplification device; and a heat insulator for blocking heat from the outside and a heat-sensing sticker capable of measuring a temperature of heat generated by the heating device, which is provided inside the storage space.

2. The portable isothermal amplification device of claim 1, wherein the heating device is selected from a hand warmer, a heating pack, and a heating pad.

3. The portable isothermal amplification device of claim 1, wherein the heating device is a solid heating composition comprising iron powder, salt, activated carbon, sawdust, vermiculite, water, or a combination thereof.

4. The portable isothermal amplification device of claim 1, wherein the heating device is a liquid heating device comprising sodium thiosulfate, sodium acetate, or a combination thereof.

5. The portable isothermal amplification device of claim 1, wherein the PCR tube includes a PCR amplification reagent for amplifying a target nucleic acid and a diagnostic reagent for detecting the target nucleic acid.

6. The portable isothermal amplification device of claim 5, wherein the target nucleic acid is a nucleic acid extracted from methicillin-resistant Staphylococcus aureus.

Description

BRIEF DESCRIPTION OF DRAWINGS

[0016] The above and other aspects, features and other advantages of the present aspects will be more clearly understood from the following detailed description taken in conjunction with the accompanying drawings, in which:

[0017] FIG. 1 shows a portable isothermal amplifier provided in exemplary embodiments of the present invention.

[0018] FIG. 2 illustrates a storage space of the portable isothermal amplification device provided in exemplary embodiments of the present invention.

[0019] FIGS. 3a and 3b show the results of experiments for detecting MRSA using the portable isothermal amplifier provided in exemplary embodiments of the present invention.

BRIEF DESCRIPTION OF EMBODIMENTS

[0020] Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings.

[0021] In addition, the terms or words used in the present specification and claims are not to be construed as being limited to their ordinary or dictionary meanings and the inventor may properly define the concept of the terms or words in order to best describe his invention. Based on the principle, the terms and words should be interpreted as meanings and concept consistent with the technical idea of the present invention.

[0022] Therefore, since the configurations shown in the embodiments and drawings described in this specification are only the most preferred embodiment of the present invention and do not represent all of the technical spirit of the present invention, it should be understood that various equivalents and modifications may be substituted for them at the time of filing the present application.

[0023] In order to diagnose infection of microorganisms or viruses in the field, such as hospitals or clinics, after obtaining a sample from the field using a tube and we should transfer it to a laboratory that detects the microorganism or virus. And after amplifying the nucleic acid contained in the tube using an isothermal PCR amplifier provided in the laboratory, we can diagnose it through a detection method such as fluorescence diagnosis.

[0024] Therefore, there is a possibility that the sample collected at the site may be contaminated or transformed during the above process. For this reason, in order to solve this problem, it is important to quickly detect the microorganism or virus at the site.

[0025] In order to solve the above problems, the portable isothermal amplification device (100) provided in exemplary embodiments of the present invention may use a PCR tube capable of amplifying a target nucleic acid contained in the sample obtained in the field.

[0026] In accordance with exemplary embodiments of the present invention, a heating device capable of maintaining a constant reaction temperature may be provided inside the portable isothermal amplifier so that the nucleic acid is efficiently amplified during the amplification process.

[0027] The portable isothermal amplification device (100) provided by exemplary embodiments of the present invention relates to a portable isothermal amplifying device capable of rapidly detecting microorganisms and viruses in a field such as a hospital or public health center.

[0028] The portable isothermal amplification device (100) in accordance with exemplary embodiments of the present invention is characterized by applying a ring-mediated isothermal amplification method (LAMP) when amplifying a nucleic acid to be detected using a PCR method.

[0029] The portable isothermal amplification device (100) in accordance with exemplary embodiments of the present invention may have a PCR tube insertion unit (20) at the top which is capable of fixing the PCR tube (10) containing the sample.

[0030] In addition, the portable isothermal amplification device (100) in accordance with exemplary embodiments of the present invention may also have a tube cover (30) for fixing the PCR tube (10) after inserting the PCR tube (10) at the top.

[0031] Additionally, in the lower part of the portable isothermal amplification device (100) provided in exemplary embodiments of the present invention, there is formed a storage space (50) for accommodating the heating device (40) such as a hand warmer for maintaining a constant reaction temperature, which is the most important condition for amplifying the nucleic acid contained in the PCR tube (10) by the LAMP method.

[0032] A storage space cover (60), which can block the storage space (50) from the outside, may be provided so that the heat generated by the heat-generating device (40) is maintained only inside the portable isothermal amplifier (100) in accordance with exemplary embodiments of the present invention.

[0033] The PCR tube (10) in the inside of the portable isothermal amplification device (100) provided in exemplary embodiments of the present invention may contain a PCR amplification reagent for amplifying a target nucleic acid such as DNA or RNA contained in the sample and a probe for detecting the target nucleic acid and diagnostic reagents composed of fluorescent substances.

[0034] Therefore, after taking a sample from the field, the sample is put into the PCR tube (10), and the tube (10) is inserted into the portable isothermal amplification device (100) provided in exemplary embodiments of the present invention. For the fluorescent solutions in the PCR tube (10), after a certain time has elapsed, there is a color change of the reagent in the tube (10) because the nucleic acid contained in the PCR tube (10) is amplified isothermally in the amplifying device (100) which amplifies the nucleic acid contained in the PCR tube (10) isothermally and reacts with the probe in the tube (10).

[0035] By using such a color change, the presence of a specific microorganism or virus contaminated in the sample can be rapidly confirmed at the field, and whether the field is infected by the microorganism or virus can be rapidly identified.

[0036] The reagent that is put into the inside of the PCR tube (10) mounted inside the portable isothermal amplification device (100) provided in exemplary embodiments of the present invention includes a PCR amplification reagent capable of amplifying any type of target nucleic acid and diagnostic reagents capable of detecting the target nucleic acid.

[0037] Therefore, the portable isothermal amplifier (100) provided in exemplary embodiments of the present invention can be applied to any kinds of microorganism or virus suspected of being infected in the field depending on the reagents injected into the PCR tube (10).

[0038] In the portable isothermal amplification device (100) provided in exemplary embodiments of the present invention, the heating device (40) for providing heat for PCR amplification of the nucleic acid of the sample contained into the PCR tube (10) may be a hand warmer, a heating pack, a heating pad, etc.

[0039] The heating device can be directly inserted into the storage space (50) provided in the portable isothermal amplification device (100) in accordance with exemplary embodiments of the present invention.

[0040] In addition, as the heating device (40), any heating composition capable of using heat-generator through oxidation of iron can be directly put into the storage space (50) and the heating composition can be a solid composition comprising iron powder, salt, activated carbon, sawdust, vermiculite, diatomaceous earth and water and the like.

[0041] The exothermic heating composition uses heat generated in the oxidation process of iron when iron powder meets oxygen and water.

[0042] In addition, as the heating composition, a liquid heating composition may be used, and the liquid composition includes sodium thiosulfate or sodium acetate and a metal piezoelectric body.

[0043] The heat generating device used in exemplary embodiments of the present invention can be any suitable heat generating device as long as it can be accommodated in the storage space (50).

[0044] FIG. 2 shows the storage space (50) of the portable isothermal amplifier (100) provided in exemplary embodiments of the present invention.

[0045] The upper drawing of FIG. 2 shows a plan view of the storage space (50), and inside of the storage space (50), there is a heat-sensing sticker (70) capable of measuring the temperature of the heating device (40) provided inside the storage space (50).

[0046] The heat-sensing sticker (70) is installed in contact with the heat-generating device (40) at the upper portion of the storage space (50), and the temperature of the heat generated from the heat-generating device (40) can be measured by the heat-sensing sticker 70.

[0047] The heat-sensing sticker (70) changes the color smoothly during the heating process. However, when the temperature exceeds a certain temperature (e.g., about 60? C.), the heat-sensing sticker (70) completely changes into another color, which can prove the isothermal amplification of the hexanes in the sample.

[0048] Such a change in temperature can be observed with naked eyes by the heat-sensing sticker (70). For example, when the heat-sensing sticker (70) which was white shows red color due to the heat generated by the heating device (40), it verifies that the isothermal amplification of the nucleic acid is actively occurring.

[0049] The lower view of FIG. 2 is a separate view of the storage space (50). An insulating material (80) is provided at the lower portion of the interior of the storage space (50), so that heat generated from the heating device is prevented from escaping to the outside. As the result, it makes sure that efficient isothermal amplification of nucleic acids is performed in the PCR tube (10).

EXAMPLES

[0050] Next, an embodiment in which methicillin-resistant Staphylococcus aureus (MRSA) is detected using the portable isothermal amplification device (100) in accordance with exemplary embodiments of the present invention will be described.

[0051] In order to detect MRSA using the portable isothermal amplification device provided in exemplary embodiments of the present invention, 25 ?l of a reagent in which the concentration of MRSA was changed was injected into the PCR tube (10) and as the exothermic heating device, a solid heat generating means made of iron powder, activated carbon, sodium chloride, vermiculite and a highly hygroscopic polymer was put into the storage space (50).

[0052] While the reaction proceeded, the change in the color of the reagent injected into the PCR tube (10) was observed at intervals of 5 minutes after the reaction was 25 minutes.

[0053] As shown in FIGS. 3a and 3b, when MRSA could be easily and quickly detected using the portable isothermal amplification device provided in exemplary embodiments of the present invention, although the concentration of MRSA was extremely small.

[0054] In FIGS. 3a and 3b, each number represents the concentration of MRSA contained in the PCR tube (10), while PC represents a positive control, NC represents a negative control.

[0055] FIG. 3a shows the state before inserting the PCR tube (10) into the portable isothermal amplifier (100), and FIG. 3b shows the time after inserting the PCR tube (10) into the portable isothermal amplifier (100) and the color change of the reagent according to the concentration of MRSA.

[0056] Although the present invention has been described with reference to the above embodiments and drawings, it is 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.