KR20200137312A - Composition for diagnosing diabetes using tear - Google Patents
Composition for diagnosing diabetes using tear Download PDFInfo
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- KR20200137312A KR20200137312A KR1020190063444A KR20190063444A KR20200137312A KR 20200137312 A KR20200137312 A KR 20200137312A KR 1020190063444 A KR1020190063444 A KR 1020190063444A KR 20190063444 A KR20190063444 A KR 20190063444A KR 20200137312 A KR20200137312 A KR 20200137312A
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Abstract
Description
본 발명은 눈물의 당 수준을 측정하여 당뇨병을 진단하는 것을 특징으로 하는 당뇨병 진단용 조성물에 관한 것이다.The present invention relates to a composition for diagnosing diabetes, characterized in that diagnosing diabetes by measuring the level of sugar in tears.
당뇨병은 혈액 내에 존재하는 포도당이 소변을 통해 배출되는 질환으로써 현대인에게 가장 많이 발생되는 비전염성 만성질환이자 근본적인 치유가 되지 않는 만성 퇴행성 질환 중의 하나이며, 인슐린 작용과 인슐린 분비에 의해 발생하는 고혈당을 특징으로 하는 대사장애 증후군으로써 자가면역기전에 의해서 췌장의 베타세포가 파괴되면서 인슐린이 절대적으로 부족하게 되는 것에서부터 인슐린의 작용에 대한 저항성에 이르기까지 다양한 원인으로 발병하게 된다.Diabetes is a disease in which glucose present in the blood is excreted through urine. It is one of the most common non-infectious chronic diseases in modern people and one of the chronic degenerative diseases that cannot be fundamentally cured. It is characterized by high blood sugar caused by insulin action and insulin secretion. As a metabolic disorder syndrome caused by autoimmune mechanisms, the beta cells of the pancreas are destroyed, resulting in a variety of causes ranging from absolute lack of insulin to resistance to the action of insulin.
이러한 당뇨병은 고혈당이 만성으로 지속되게 하면서 당질대사 뿐만 아니라 지질이나 단백질 대사장애를 유발하게 하여 망막, 신장, 신경, 심혈관계 등의 합병증을 유발시키는 심각한 질병이며 다양한 합병증으로 인해 수명이 5~10년 정도 단축될 수 있고, 현재 우리나라에서도 사망원인 중 4번째의 질병이다.Such diabetes is a serious disease that causes complications such as retina, kidney, nerve, cardiovascular system, etc. by causing not only glucose metabolism, but also lipid or protein metabolism, while maintaining high blood sugar chronically. It can be shortened, and is currently the fourth disease among the causes of death in Korea.
최근 급속한 경제발전에 따른 식생활의 변화로 당뇨병 유병율은 해마다 증가하여 국내의 경우 약 5~10%에 달하고 있으며, 당뇨병 환자는 급속히 증가하고 있는 반면, 지속적이고 적절한 치료가 어려운 상황이기 때문에 이런 당뇨병을 미리 진단하고 꾸준한 관리할 수 있는 방법이 중요하다.Due to the recent rapid economic development and dietary changes, the prevalence of diabetes has increased year by year, reaching about 5-10% in Korea, and the number of diabetic patients is increasing rapidly, but it is difficult to treat such diabetes in advance. It is important to be able to diagnose and manage consistently.
가장 많이 사용되는 당뇨검사 키트는 채혈침을 사용하여 손가락 끝의 혈액을 이용해 자가진단하는 방법으로 이러한 방법은 매 검사시 신체에 상처를 입혀야하고 고통이 수반되는 단점이 있다. 이에 따라 비침습적인 방법으로 환자의 고통을 최소하하며 정확한 당 수치를 측정할 수 있는 당뇨 진단 방법에 대한 개발이 필요하다.The most commonly used diabetes test kit is a method of self-diagnosis using blood from the tip of a finger using a blood collection needle. This method has the disadvantage of injuring the body during each test and accompanying pain. Accordingly, there is a need to develop a diabetic diagnosis method capable of minimizing patient pain and measuring accurate glucose levels through a non-invasive method.
본 발명은 기존의 당뇨 진단법의 문제점을 해결하기 위하여 눈물 속 당을 활용하여 비침습적인 방법으로 당 수치를 측정하여 당뇨병을 진단하기 위한 조성물을 제공하고자 한다.An object of the present invention is to provide a composition for diagnosing diabetes by measuring sugar levels in a non-invasive method using sugar in tears in order to solve the problems of the existing diabetes diagnosis method.
본 발명은 눈물의 당 수준을 측정하여 당뇨병을 진단하는 것을 특징으로 하는 당뇨병 진단용 조성물을 제공한다.The present invention provides a composition for diagnosing diabetes, characterized in that diagnosing diabetes by measuring the level of sugar in tears.
본 발명에 따르면, 기본적인 효소 반응을 이용하여 색 변화를 통한 눈물 내 포도당 검출에 있어서, 효소와 함께 알지네이트를 포함시킨 조성물은 종래 효소 반응을 이용한 당뇨병 진단 방법보다 눈물 내 저농도의 포도당까지 뚜렷한 색변화를 통하여 검출하는 것으로 확인됨에 따라, 상기 조성물은 눈물을 이용하여 간편하고 비침습적인 당뇨 진단용 시약 및 진단용 키트로 제공될 수 있다.According to the present invention, in the detection of glucose in tears through color change using a basic enzymatic reaction, a composition containing alginate together with an enzyme exhibits a distinct color change to a low concentration of glucose in tears than a conventional diabetic diagnosis method using an enzymatic reaction. As it is confirmed through detection, the composition may be provided as a simple and non-invasive diabetic diagnostic reagent and a diagnostic kit using tears.
도 1은 포도당 측정을 윈한 주요 효소 반응을 나타낸 모식도이다.
도 2는 기본적인 효소 반응을 활용한 포도당 농도별 색 변화를 확인한 결과이다.
도 3은 알지네이트 첨가를 통하여 포도당 농도별 색 변화를 확인한 결과이다.
도 4는 다양한 알지네이트 농도에서 포도당 검출 색 변화를 확인한 결과이다.
도 5는 포도당 농도에 따른 키토산과 알지네이트 색 변화를 확인한 결과이다.1 is a schematic diagram showing the main enzyme reactions for glucose measurement.
2 is a result of confirming the color change for each glucose concentration using a basic enzymatic reaction.
3 is a result of confirming the color change by glucose concentration through the addition of alginate.
4 is a result of confirming the color change of glucose detection at various alginate concentrations.
5 is a result of confirming the color change of chitosan and alginate according to the glucose concentration.
이하, 본 발명의 이해를 돕기 위하여 실시예를 들어 상세하게 설명하기로 한다. 다만 하기의 실시예는 본 발명의 내용을 예시하는 것일 뿐 본 발명의 범위가 하기 실시예에 한정되는 것은 아니다. 본 발명의 실시예는 당업계에서 평균적인 지식을 가진 자에게 본 발명을 보다 완전하게 설명하기 위해 제공되는 것이다.Hereinafter, examples will be described in detail to aid understanding of the present invention. However, the following examples are for illustrative purposes only, and the scope of the present invention is not limited to the following examples. The embodiments of the present invention are provided to more completely explain the present invention to those of ordinary skill in the art.
<실시예 1> 포도당 측정의 기본원리 및 종이 선정<Example 1> Basic principle of glucose measurement and paper selection
당을 측정하기 위한 기본 원리는 도 1과 같이 포도당과 포도당 산화효소와의 반응, 상기 반응의 산물인 과산화수소와 과산화효소, 발색 시약(Dye)과의 반응이다.The basic principle for measuring sugar is a reaction between glucose and glucose oxidase, as shown in FIG. 1, and a reaction between hydrogen peroxide and peroxidase, which is a product of the reaction, and a color developing reagent (Dye).
먼저, 기본적인 효소만을 사용하여 포도당 농도 별 색 변화를 관찰하기 전에 키트 개발에 사용될 3가지 후보의 종이 중 눈물을 쉽게 흡수할 수 있는 능력을 확인할 수 있는 특징인 종이의 미세한 구멍 크기와 물을 흡수하는 속도를 확인하였다.First, before observing the color change by glucose concentration using only basic enzymes, among the three candidate papers to be used in the kit development, it is possible to confirm the ability to easily absorb tears. Check the speed.
그 결과, 표 1과 같이 미세 구멍의 크기가 크고 물 100ml을 흡수하는데 걸리는 시간이 짧은 Whatman사의 filter paper No.41을 선정하였다.As a result, as shown in Table 1, filter paper No. 41 of Whatman, which has a large pore size and a short time to absorb 100 ml of water, was selected.
<실시예 2> 포도당 색 변화 검출을 위한 조성물 확인<Example 2> Confirmation of the composition for detection of glucose color change
1. 효소를 사용한 포도당 농도 별 색 변화 확인1. Check the color change by glucose concentration using enzyme
기본적인 포도당 측정 방법인 포도당 산화효소와 과산화효소를 사용하고, 발색 시약으로 TMB(3,3',5,5'-Tetramethylbenzidine) 사용하여 포도당 농도에 따른 발색 시약(TMB)의 색 변화 여부를 확인하였다.Glucose oxidase and peroxidase, which are basic glucose measurement methods, were used, and TMB (3,3',5,5'-Tetramethylbenzidine) was used as a color developing reagent to check the color change of the color development reagent (TMB) according to the glucose concentration. .
그 결과, 도 2와 같이 2번의 실험에서 포도당 농도가 높아짐에 따라, 색의 변화는 뚜렷하게 나타났지만, 농도가 낮은 0.1 nM에서의 색 차이를 구별하기는 어려웠다.As a result, as the glucose concentration increased in the second experiment as shown in FIG. 2, the color change was clearly observed, but it was difficult to distinguish the color difference at 0.1 nM with a low concentration.
2. 알지네이트(Alginate)를 첨가한 포도당 농도 별 색 변화 확인2. Check the color change according to the glucose concentration added with alginate
기본적인 효소 반응을 활용하여 포도당 농도별 색 변화를 확인한 결과, 저농도에서의 색 변화는 확인이 어려웠으며, 이를 해결하기 위해 여러 첨가물질을 조사하던 중, 효소 고정화 능력이 있는 알지네이트를 2% 및 5% 농도로 사용하여 포도당 농도별 색 변화를 확인하였다.As a result of confirming the color change by glucose concentration using a basic enzymatic reaction, it was difficult to confirm the color change at low concentration, and while investigating various additives to solve this problem, alginate with enzyme immobilization ability was 2% and 5%. It was used as a concentration to check the color change by glucose concentration.
그 결과, 도 3과 같이 알지네이트를 첨가한 실험군의 색변화가 기본적인 반응만을 사용한 대조군보다 색 변화가 뚜렷하고 저농도에서도 색 변화가 분명하게 나타났다. As a result, as shown in FIG. 3, the color change of the experimental group to which alginate was added was more pronounced than the control group using only the basic reaction, and the color change was evident even at low concentration.
상기 결과로부터 효소 고정화 능력이 있고 색 변화를 뚜렷하게 만들어 주는 알지네이트를 당 측정 키트의 보조물질로 선정하고, 알지네이트의 최적의 알지네이트 농도를 확인하기 위해, 포도당 농도를 0.5mM로 고정하고 알지네이트의 농도 별 색 변화를 확인하였다.From the above results, alginate, which has enzyme immobilization ability and makes color change distinct, was selected as an auxiliary material of the sugar measurement kit, and in order to check the optimal alginate concentration of alginate, the glucose concentration was fixed at 0.5 mM and the color of each alginate concentration Confirm the change.
그 결과, 도 4와 같이 알지네이트 농도가 변함에 따라 색 변화는 나타나지 않았다. 다만 알지네이트 농도가 높아짐에 따라 물에 쉽게 녹지 않고 점성이 높아져 종이에 스며들어 퍼지지 못하고 뭉쳐 있는 경향이 확인되었다.As a result, color change did not appear as the alginate concentration was changed as shown in FIG. 4. However, as the alginate concentration increased, it was found that it was not easily dissolved in water and became more viscous, so that it did not penetrate into the paper and could not spread.
상기 결과로부터 고농도의 알지네이트는 부적합한 것이 확인됨에 따라, 물에 잘 용해되고 점성이 높지 않은 2% 알지네이트가 가장 적합한 것을 확인할 수 있었다.From the above results, as it was confirmed that alginate at a high concentration was unsuitable, it was confirmed that 2% alginate, which is well soluble in water and not high in viscosity, is most suitable.
<실시예 3> 포도당 농도에 다른 키토산 및 알지네이트 성능 비교<Example 3> Comparison of chitosan and alginate performance in different glucose concentrations
앞서 보고되어진 키토산과 알지네이트와의 성능을 비교하기 위해 도 5와 같이 여러 농도의 포도당을 사용하여 색 변화를 확인하였으며, 색을 분석할 수 있는 모델 중 하나인 RGB와 HSV에서 R 값과 S 값만을 이용하여 각각의 포도당 농도에서의 색을 값으로 변환하여 분석하였다.In order to compare the previously reported performance of chitosan and alginate, as shown in FIG. 5, color change was confirmed using glucose of various concentrations, and only R and S values in RGB and HSV, which are one of the models capable of analyzing color, The color at each glucose concentration was converted into values and analyzed.
그 결과 표 2와 같이 모바일 알고리즘을 고려하여 포도당 앞 농도와의 값 차이를 확인한 결과, R 값을 활용하여 분석하였을 때 가장 적합한 것으로 확인되었다.As a result, as shown in Table 2, as a result of confirming the difference in the value from the concentration in front of glucose in consideration of the mobile algorithm, it was found to be the most suitable when analyzed using the R value.
이상으로 본 발명 내용의 특정한 부분을 상세히 기술하였는 바, 당업계의 통상의 지식을 가진 자에게 있어서, 이러한 구체적 기술은 단지 바람직한 실시양태일 뿐이며, 이에 의해 본 발명의 범위가 제한되는 것이 아닌 점은 명백할 것이다. 따라서 본 발명의 실질적인 범위는 첨부된 청구항들과 그것들의 등가물에 의하여 정의된다고 할 것이다.As described above, specific parts of the present invention have been described in detail, and for those of ordinary skill in the art, it is obvious that these specific techniques are only preferred embodiments, and the scope of the present invention is not limited thereby. something to do. Therefore, it will be said that the practical scope of the present invention is defined by the appended claims and their equivalents.
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| KR20220125621A (en) | 2021-03-05 | 2022-09-14 | 경희대학교 산학협력단 | Method and kit for diagnosing diabetes using tear samples |
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| KR20170002318A (en) | 2015-06-29 | 2017-01-06 | 고려대학교 산학협력단 | Composition for early diagnosing diabetes using metabolomics |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20220125621A (en) | 2021-03-05 | 2022-09-14 | 경희대학교 산학협력단 | Method and kit for diagnosing diabetes using tear samples |
| KR20230038173A (en) | 2021-03-05 | 2023-03-17 | 경희대학교 산학협력단 | Method and kit for diagnosing diabetes using tear samples |
| KR20230038174A (en) | 2021-03-05 | 2023-03-17 | 경희대학교 산학협력단 | Method and kit for diagnosing diabetes using tear samples |
| KR20230038172A (en) | 2021-03-05 | 2023-03-17 | 경희대학교 산학협력단 | Method and kit for diagnosing diabetes using tear samples |
| KR20230038681A (en) | 2021-03-05 | 2023-03-21 | 경희대학교 산학협력단 | Method and kit for diagnosing diabetes using tear samples |
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