M383733 五、新型說明： 【新型所屬之技術領域】 本創作係有關於一種糖化血色素比率測量裝置，尤指一種用 於計算糖化血色素(HbAlc)比率的連續式免疫分析定量裝置，包含 —反應管，可容納反應物及反應試劑於其内作用，以及一反應管 槽，其中之反應管槽底部置有磁性槽座，側邊兩端設有量測視窗， 以量測反應管中反應物之吸光值。 【先前技術】 血色素是紅血球内所含的一種蛋白質，為兩個一樣次單位蛋 白構成二聚體並進一步將兩個二聚體組成一異四聚體(heter〇tetramer) ，其主要功能是將氧氣帶到組織並將二氧化碳帶離組織。由於葡 萄糖可自由通過紅血球，所以血液中葡萄糖可經由可逆非酵素方 式與血色素結合，伴隨血中葡萄糖濃度與結合時間等因子之影響 衍生出不可逆的骑化血色素分子，直到紅血球達生命期而自 行裂解為止。因此每個紅血球内的少數血色素分子都有葡萄糖以 可逆或不可逆型式結合，#巾特定結合在▲色素料單位蛋白N 端的綠胺酸(VaHne)的血色素分子便稱為糖化血色素⑽Ak)。一 旦血液中葡萄糖濃度愈高時，則會有愈多靴▲色素分子，所以 糖化血色姐轉㈣之提昇。tg躲與血色素結合並轉變成 不可逆型式後，會-直以聽蛋白模式存在直到紅血球裂解為止 ’而紅血球解均壽麵四個月，再加上造·萄姆合血色素 3 M383733 至不可逆型式之機制是伴隨血糖濃度與作科間長鱗因素，因 此糖化A色·比率可充分反映出紅血球生存__平均血糖 濃度。即便在正常血糖濃度魏下血色素也會生成不可逆型式之 醣化蛋白，根職國絲病學會猶議—般人的糖化血色素正常 比率約在4-6%間。 目别已開發出用於檢聰化血色素比率的方法及I置約有3〇 ►多種’按照制原理可大賴分為兩大類H基於糖化血色 素與非糖化血色素分子上電荷的不同，再藉由電荷差異所導致在 官柱中靜相與流動相間移動的分佈差異加以分離之^^離子交 換層析和m-是基於血色素上是祕合葡雜所產生的結 構差異’再_可區分技有結合賴歡狀分子加以分離之 如親和層析和免疫分析。其中離子交換液相層析法是利用錄 素因糖化與否導致其帶紐有所的柯，於是樣品在通過離子交 換的分析管_，便會按攜帶電荷的差異於㈣濃麟度的緩衝 液刀別冲提出達到分離及分析定量目I至於硼酸⑻士 _) ^合力層析法也是彻管柱層析法，但所_是瓣會與糖類功 能基tos-diol)間產生特殊親和作用力結合糖化血紅素，因此所 備測的對象是所有被糖化的血色素，也就是說僅可區分血紅素是 有、。。酶類’而無法進一步確認該醣化蛋白是否就是糖化血色 素二雖然上述兩種檢測方法已廣泛被使収準確程度也符合量測 之需求’但執行量測時皆需在控制良好的環境下操作，以致無法 4 M383733 發展出小型可攜帶賴組而局限其應雜。除此之外，兩方法雖 然都可重複使用，但該的特色在維護不當下反而會產生量測結果 偏差更大的後果。至於硼酸親合力層析法因檢測原理無法直接獲 得糖化蛇素值’而需代人轉換公式求得之糖化血色素值。針對 上述量測方法之缺失，以免疫檢_則關發的糖化血色素比率 定量系統皆可有效克服，但以免疫檢測技術應用在糖化血色素比 率h_L仍有_量_理之需求所導致設計料（縣過高）及 準確性較差等若干缺點’躺影響免疫檢職術歧地應用在糖 化血色素比率定量上的可行性。 少數以免疫_技術關發定化血色纽率的商品目前 已知的有兩種’其-是採免疫比濁法取糖化血色素而定量糖 化血色素瞬則是應轉殊設計含試舰，如美國專利議5〇93 齡’在搭配已鍵人可提供自動化操作程序之量測設備下，使反應作 用可依序執狀。錢程如下：先制雜高鐵錄素比色法洌 定檢體帽血色素蛋^私競爭纽關法浙铺t糖化血 色素針對測疋糖化血色素部分，當紅血球被破壞並釋放血色素 後八中糖化血色素在與含有單株抗體之試劑進行特異結合，同 時試舰亦·有已缝姉微粒之娜化血色素分子複合物可 與單株抗體結合並產生_反應，由於血中糖化血色素會使得反 應槽内的雜毅下降，於是反應溶液财賴化減本中糖化 血色素的濃度成反比。最後再藉由已内鍵以驗性高鐵血紅素比色 5 M383733 法制定之總血色素濃度標準曲線及以競爭免疫比濁法制定之糖化 血色素濃度鮮曲線，分別計算出受齡血樣品中總血色素及糖 化血色素濃度’進而求得糖化血色素比率值。 另一已上市以免疫分析定量糖化血色素比率的商品，是應用 -傳統免疫偵測技術搭配微光學技術(MiCr〇-opticaItechn〇i〇gy)測定 糖化A色素的比率，主要技術是顧對血色素及糖化▲色素分子 •具專-結合能力之抗體，再分別標示不同的螢光物質，並藉由特 殊設計側流層析(lateralflow)裝置，如美國專利卿98聰，以達 到可平均分流受測樣品之方式，以便於可在同時間分別檢測出待 測物中總血色素及糖化金色素的含量，再搭配極精密之微光學技 術裝置戴取側流層析裝置上所呈現與總血色素及糖化血色素含量 成正比的訊號：¾後根據已内鍵入的總血色素及糖化血色素濃度 ‘準曲線’便可分別計算出受測全血樣品#總血色素及糖化血色 素〉辰度，進而求得糖化血色素比率值。 雖然已開發ib檢聰化血色素比率的方法及裝置，且有已上 市之商品採収疫侧技術酬糖化血色素比率，但是採取免疫 分析技術從事糖化血色纽率之測量傾分難得總血色素及糖 化血色兩者個別在同—待測物中之濃度。而任何量測技術應用繁 雜叹计並導人自動倾式，仍無法完全排_職不精確性，況 6 M383733 且目前以免疫偵測技術模式測量糖化血色素比率時都必須針對總 血色素及糖化血色素在同一待測物中之濃度分別執行檢洌，因此 在一模式下的不準確程度勢必遠高於其他方法。 -般人正常糖化血色素的正常值約在4_6%間，而糖尿病患者 的糖化喊雜也鮮㈣在_下，這錢找麵量糖^血 色素時所檢測總血色素及糖化血色素在訊號上有可能產生十倍以 上的差異’為了能同時解析並調和過高或過低訊號，就必須減損 對兩者量咖準確度，再加上所欲量贱糖化血色素在總灰色素 _率值，所时受計算雜域制之不準雜，進而導致測 量結果失真。本難即針對先前技術之不足，包括為了檢測糖化 血色素在總血色素的比率值所料人f瑣設計而造成過高材料成 本'使用不便捷及量測之不準確性等缺憾，開發出義免疫偵測 技術之糖化血色素比率測量方式。 【新型内容】 傳統以免疫侧技蚊化A色素比率的裝置大致可區分 成兩大類，其原理分贱應用抗體對標的物具專-辨識結合之特 性，並對待測樣品t血色素及糖化血色素各自進行量測；或是僅 使用抗體檢測糖化血色素含量在搭配已知之血色素定量方法下， 再計算出糖化血色素的比率。至於測量的方法及裝置則視檢測模 式採取不_型，糾抗财量槪血色敍血色鱗，則多藉 7 M383733 由抗體上的標定物，如：螢光物質或酵素，按結合標的物的抗體 分子之多寡，產生與其濃度成正相關的訊號，最後再計算出血樣 中糖化血色素的比率。在以光學方法定量總血色素是測量與反應 試劑作用後血色素在特定波長下的吸光值(〇ptical density)，至 於所搭配的免疫分析則是利用抗體-抗原複合體生合成，再搭配可 增強訊號的微粒子便可以光學方法量測糖化血色素濃度。請參閱 第一圖，本創作所採用適用於本裝置的試劑係以具官能基修飾的 籲磁性跡以共舰結合至抗糖化血色素分子抗體，製作成生物專 一性磁珠，一方面藉由抗體所具有專一辯識結合特性做為偵測工 具；另一方面在搭配可提供磁力裝置下，利用生物專一性磁珠分 離之，並藉以收集已結合收集裝置之糖化血色素。 請參閱第二圖，本創作之糖化血色素比率測量裝置包含一反 .應管，可谷納反應物及反應試劑於其内作用，以及一可提供做為 執行分離_之反鮮槽，除了反鮮本身可制在鮮量測上 ，並提供符合執行吸光值測量之需求，反應管槽亦供光學量測執 行之區域是水平且穩固。m，反應管所裝填之反應試劑包 含三種：一為溶血素，提供可分解紅血球之成分，如此一來當待 測樣品（全血)加入後，紅血球遭到破壞並釋出其中的血色素，亦 包括糖化血色素在内；另一反應試劑則為血色素定量試劑，其與 血色素結合’進而強化血色素在特定波長下吸光值，以便於定量 企色素浪度，以及如第二圖所示之生物專一性磁珠，由於以共價 8 M383733 結合了單株或多株之抗糖化血色素抗體，可專—性_糖化血色 素結合，受_性做㈣導至反絲底.反應f槽有兩特殊 設計’其-是位於反鮮槽侧邊兩端並可讓_光_利通過反 應管槽的制視窗，當執行血色诚度量測時，包括糖化血色素 濃度，特定波長光源便可透過管槽側邊量測視窗完狂色素（包括 糖化也色素)濃度量測。另-是錄反絲触部驗提供磁力的 磁性槽座，於是當反應管内有收集糖Μ色錄置並置於反應管 槽時，則底部磁性槽座便會將結合糖化&色素的複合物吸至反應 管底部。鱗透過本齡之糖化i色纽率測量裝置可將所量測 排除糖化血色素之血色魏度，整合兩:欠量聽果並配合修改過 的糖化血色素畔計算方程式，便可獲得酬樣品帽化血色素 佔總血色素百分比。詳細·計算方式如下：_後於特定波長 下執行量騎得血色素财A值；加Μ物專—性磁珠至反應管 卜使生物專-性磁珠與待測檢體中糖化血㈣結合，並透過磁 性槽座將與生物專-性磁珠結合之糖化血色素吸至反應管底部 於設定的特定波長下執行量晴得衫奴度β ;以及將A 與B帶入下述公式M383733 V. New description: [New technical field] This creation is about a glycosylated hemoglobin ratio measuring device, especially a continuous immunoassay quantitative device for calculating the ratio of glycated hemoglobin (HbAlc), including - reaction tube, The reaction medium and the reaction reagent can be accommodated therein, and a reaction tube tank is provided, wherein a magnetic tank seat is arranged at the bottom of the reaction tube groove, and measuring windows are arranged at both ends of the reaction tube to measure the light absorption of the reactants in the reaction tube value. [Prior Art] Hemoglobin is a protein contained in red blood cells, which constitutes a dimer of two identical subunit proteins and further forms two dimers into a heterotetramer (heter〇tetramer) whose main function is to Oxygen is brought to the tissue and the carbon dioxide is carried away from the tissue. Since glucose can pass freely through red blood cells, glucose in the blood can be combined with hemoglobin via a reversible non-enzymatic method, and irreversible riding of hemoglobin molecules can be derived from factors such as blood glucose concentration and binding time, until the red blood cells reach the life stage and lyse themselves. until. Therefore, a small number of hemoglobin molecules in each red blood cell have glucose combined in a reversible or irreversible manner. The hemoglobin molecule of phytic acid (VaHne) specifically bound to the N-terminus of the ▲ pigment unit protein is called glycated hemoglobin (10) Ak). Once the blood glucose concentration is higher, there will be more ▲ pigment molecules, so the glycosylation of the blood color sister (4) is improved. After tg hides and binds to hemoglobin and transforms into an irreversible pattern, it will be in the form of listening to the protein until the red blood cells are lysed, and the red blood cell is treated for four months, plus the mechanism of making the hemoglobin 3 M383733 to the irreversible pattern. Along with the blood glucose concentration and the long scale factor between the subjects, the saccharification A color ratio can fully reflect the red blood cell survival __ average blood glucose concentration. Even at normal blood glucose levels, hemoglobin produces an irreversible type of glycated protein, and the Root National Silk Disease Society believes that the normal ratio of glycated hemoglobin is about 4-6%. The method for detecting the hemoglobin ratio has been developed and I have about 3 〇 ► a variety of 'according to the principle of the system can be divided into two major categories of H based on the difference in the charge of glycated hemoglobin and non-glycated hemoglobin molecules, and then borrow The separation of the distribution of the movement between the stationary phase and the mobile phase caused by the difference in charge is separated by ion exchange chromatography and m- is based on the structural difference produced by the hemiglobin on the hemoglobin. There are binding to Laihuan molecules for separation such as affinity chromatography and immunoassay. Among them, ion exchange liquid chromatography is to use the pheromone due to saccharification or not to cause its kinetics, so the sample is in the analysis tube through ion exchange, and it will be different according to the carrying charge (4) dense cytoplasmic buffer. The knife is proposed to achieve separation and analysis of quantitative I. To boric acid (8) Shi _) ^ Synergy chromatography is also a column chromatography, but the _ is the valve and the sugar functional group tos-diol) to produce a special affinity In combination with glycated hemoglobin, the object to be tested is all glycated hemoglobin, that is to say, only hemoglobin can be distinguished. . Enzymes' cannot further confirm whether the glycated protein is glycated hemoglobin. Although the above two detection methods have been widely used to meet the measurement requirements, the measurement needs to be performed in a well-controlled environment. As a result, 4 M383733 could not develop a small portable group and it was limited. In addition, although the two methods can be reused, the characteristics will result in more deviations in measurement results due to improper maintenance. As for the boric acid affinity chromatography method, the glycated venom value cannot be directly obtained due to the detection principle, and the glycated hemoglobin value obtained by the substitution formula is required. In view of the above-mentioned lack of measurement methods, the glycosylated hemoglobin ratio quantification system can be effectively overcome by immunoassay, but the application of immunoassay technology in the glycated hemoglobin ratio h_L still results in the design of materials ( The county is too high) and the accuracy is poor, and so on. The effect of lying on the quantitative determination of glycosylated hemoglobin ratio is affected. A small number of products that use the immune-technical shutdown to determine the color of the blood color are currently known to have two kinds of 'these--the immunoturbidimetric method for taking glycated hemoglobin and the quantitative glycosylated hemoglobin is a design that should be changed to include a test ship, such as the United States. The patent proposal 5〇93 age' is used in the measurement equipment that can provide automatic operation procedures with the key person, so that the reaction can be executed in order. Qian Cheng is as follows: First, the production of high-speed iron recording colorimetric method, the detection of cap blood pigmented eggs, the private competition, the method of Zhepu, the glycosylation of hemoglobin, and the hemoglobin of the sputum, when the red blood cells are destroyed and the hemoglobin is released. It specifically binds to the reagent containing the monoclonal antibody, and the naval hemoglobin molecule complex with the spliced sputum particles can bind to the monoclonal antibody and produce a _reaction, which is caused by glycosylated hemoglobin in the blood. The amount of hydration decreased, so the concentration of glycated hemoglobin in the reaction solution was inversely proportional. Finally, the total hemoglobin concentration curve of the age-appropriate blood sample was calculated by the standard curve of the total hemoglobin concentration established by the method of the intrinsic methemoglobin colorimetric 5 M383733 method and the fresh curve of the glycated hemoglobin concentration by the competitive immunoturbidimetric method. And the glycated hemoglobin concentration' to obtain the glycated hemoglobin ratio value. Another commodity that has been marketed to quantify the ratio of glycated hemoglobin by immunoassay is the application of traditional immunodetection technology combined with micro-optical technology (MiCr〇-opticaItechn〇i〇gy) to determine the ratio of glycated A pigment. The main technique is to consider hemoglobin and Glycosylation ▲ pigment molecules • antibodies with specific-binding ability, respectively, labeling different fluorescent substances, and by special design lateral flow chromatography (lateralflow) device, such as US Patent Secretary 98 Cong, to achieve average shunt measurement The sample is so as to be able to detect the total hemoglobin and glycosylated gold content in the test object at the same time, and then with the ultra-precise micro-optical device to wear the side stream chromatography device and the total hemoglobin and saccharification. The hemoglobin content is proportional to the signal: after 3⁄4, the total hemoglobin and glycated hemoglobin concentration 'quasi-curve' can be calculated to calculate the total hemoglobin and glycated hemoglobin>, and then determine the glycated hemoglobin ratio. value. Although the method and device for detecting the hemoglobin ratio of ib have been developed, and there is a ratio of the glycosylated hemoglobin ratio of the commercially available epidemic technologies, the immunoanalytical technique is used to measure the glycosylation rate. The total hemoglobin and glycosylation are rare. The concentration of the two is in the same - the analyte. However, any measurement technology application is complicated and leads to automatic tilting. It is still impossible to fully align the inaccuracy. Condition 6 M383733 and currently measuring the glycated hemoglobin ratio in the immunodetection technology mode must be directed to total hemoglobin and glycated hemoglobin. The concentration in the same test object is separately checked, so the degree of inaccuracy in one mode is bound to be much higher than other methods. - The normal value of normal human glycosylated hemoglobin is about 4_6%, while the saccharification of diabetic patients is also fresh (4). Under the _, it is possible to detect the total hemoglobin and glycated hemoglobin in the signal. Produce a difference of more than ten times. In order to be able to simultaneously analyze and reconcile too high or too low a signal, it is necessary to detract from the accuracy of the two, and add the desired amount of glycated hemoglobin in the total gray pigmentation rate. The calculation of the miscellaneous system is not allowed to be mixed, which leads to distortion of the measurement results. This difficulty is aimed at the shortcomings of the prior art, including the development of the immunization for the purpose of detecting the ratio of the glycated hemoglobin in the total hemoglobin ratio, resulting in excessive material cost, inconvenience in use and inaccuracy in measurement. Detection technique for measuring the glycated hemoglobin ratio. [New content] Traditionally, the device that uses the immune side technology to mosquito A-pigment ratio can be roughly divided into two categories. The principle is to apply the antibody-specific object to the specific identification-binding characteristics, and to measure the sample t-hemoglobin and glycated hemoglobin. The measurement is performed; or the antibody is used to detect the glycated hemoglobin content, and the ratio of glycated hemoglobin is calculated by using the known hemoglobin quantification method. As for the method and device for measurement, depending on the detection mode, the type of the test is taken, and the amount of the blood and the color scale is used to rectify the amount of the blood, and the 7 383733 is used to calibrate the antibody, such as a fluorescent substance or an enzyme, according to the combination of the target. The amount of antibody molecules produces a signal that is positively correlated with its concentration, and finally the ratio of glycated hemoglobin in the blood sample is calculated. The quantitative determination of total hemoglobin by optical means is to measure the absorbance of hemoglobin at a specific wavelength after the action of the reaction reagent, and the immunoassay is to synthesize the antibody-antigen complex, and then match the enhanced signal. The microparticles can be used to optically measure the glycated hemoglobin concentration. Referring to the first figure, the reagents suitable for the device used in the present invention are functionally modified by a magnetically modified magnetic field to bind to an anti-glycated hemoglobin molecule antibody to prepare a biospecific magnetic bead. It has a unique combination of distinguishing characteristics as a detection tool; on the other hand, it is separated by bio-specific magnetic beads in combination with a magnetic device, and collects glycated hemoglobin of the combined collection device. Referring to the second figure, the glycated hemoglobin ratio measuring device of the present invention comprises a reverse tube, a reaction of the glutamic acid reactant and the reaction reagent, and a counter tank which can be provided as a separation separation. Fresh itself can be made on the freshness measurement and provides the requirement to perform the measurement of the absorbance value. The area where the reaction tube groove is also used for optical measurement is horizontal and stable. m, the reaction reagent filled in the reaction tube comprises three kinds: one is hemolysin, and provides a component capable of decomposing red blood cells, so that when the sample to be tested (whole blood) is added, the red blood cells are destroyed and the hemoglobin is released, and Including glycated hemoglobin; another reaction reagent is a hemoglobin quantification reagent, which binds to hemoglobin' to enhance the absorbance of hemoglobin at a specific wavelength, in order to quantify the pigmentation wave, and the biospecificity as shown in the second figure. Magnetic beads, due to the combination of a single or multiple strains of anti-glycated hemoglobin antibodies at a covalent price of 8 M383733, can be combined with _ glycosylated hemoglobin binding, by _ sex (four) lead to the anti-silk bottom. Reaction f-slot has two special designs' It is a window located at the opposite sides of the anti-fresh tank and allows the _light_ to pass through the reaction tube groove. When performing the blood color measurement, it includes the glycated hemoglobin concentration, and the specific wavelength light source can pass through the side of the tube groove. Measurement of the concentration of mad pigments (including saccharification and pigmentation) in the window. In addition, it is a magnetic magnetic socket that provides a magnetic force to the reverse silk contact. When the reaction tube has a collection of sugar and is placed in the reaction tube, the bottom magnetic housing will combine the glycation & pigment complex. Aspirate to the bottom of the reaction tube. The scales can be used to measure the blood color of the glycated hemoglobin through the saccharification i-color ratio measuring device of the same age, and integrate the two: the under-harvesting fruit and the modified glycated hemoglobin calculation equation, and the reward sample cap can be obtained. Hemoglobin accounts for the percentage of total hemoglobin. The detailed calculation method is as follows: _ after the execution of the specific wavelength, the hemoglobin A value is added; the sputum-specific magnetic beads are added to the reaction tube to combine the bio-specific magnetic beads with the glycated blood (4) in the sample to be tested. And through the magnetic socket, the glycated hemoglobin combined with the bio-specific magnetic beads is sucked to the bottom of the reaction tube at a specific wavelength to perform a smattering degree; and A and B are brought into the following formula
Hb Ale % = (A-B)/a^i〇〇 即可得到糖化血色素比率。 與現行採免疫偵測技術定量糖化血色素 創作在執行_谢輪邮__ •• 本 9 M383733 1.傳統採免疫檢測技術定量糖化血色素比率之習見裝置，其在針 對量測標的物’包括：總血色素及糖化企色素，有的是對兩者皆 使用對其具專-性之抗體進行量測；亦或是對其―，例如：使用 抗糖化血色素抗體，使用抗體進行偵測，再對另—制標目使用 其匕非免疫方式，例如：在特定波長下進行吸光度測量之光學分 析。無論是都使用或是僅對其—使用抗體做為勤江具，f尤應用 免疫檢測技術執行量測糖化血色素比率上是需制時使用到兩種 籲測量分析模式。也由於最終需納入上述兩標的測量結果，方能計 算出每一個體真正地糖化血色素比率（因每位受試者血色素的含 董有所差異）。因此’當使用免疫檢測技術定量糖化血色素比率 時，便會因整合兩種類型或是雖使用相同類型但整合兩不同偵測 工具時’導致#算所得之糖化血色素比率值易產生較大的變異， 進而導致欲採用免疫檢測技術開發定量糖化血色素比率產品有所 •阻礙i於本創作疋藉由導人生物專—(丨生磁珠成為-標記偵測裝 置，在磁性槽座下，便能發揮其特性將反應管内糖化血色素分子 加以I集至反應管底部近磁性槽座處。此時以光學比色裝置執行 里測時，所檢測出的血色素濃度則是已扣除糖化血色素之血色素 濃度值。另一方面’以光學比色農置在未加入糖化血色素抗體一生 物專一性磁珠複合體時執行量測，在搭配已知濃度的運算方程式 計算出總血色素濃度。更進―步導人計算方程式，可獲得待測物 中的糖化jk色素比率值。如H在同—彳貞測方法模式下，將 可排除免疫檢測技術在定4糖化金色素比料因_時使用到兩 10 M383733 所可能 兩種測量分龍式，無論是縣糾歧_檢測方法 導致最終量測結果有過大變異之現象。 之裝 2·由於採取免疫檢測技術朗於傳統定量糖化血色素 讀化血色素轉之概，而需針躲樣檢體在相 问反應程序下_執行糖化血色讀血色素濃度麵，勢必 輕計搭配_操作流程崎耻述㈣。如此這樣—來必定導 致製造成本增加及制獨確料良縣，她摘作可於常規 使用的光學比色管執行操作，藉助磁性槽座的特殊設計便可 達到分離彙化血色素分子，再應祕配上獅作程序之計算 方，式’便可十分容易獲知檢測樣品中糖化血色素的比率，而且 估算整體讀料成本亦遠傭财同_之商品。 3.已知-般人糖化蛇素的正常值約在4_哪間，而糖尿病患者 之糖化血色素值也盡可能㈣在_τ。但是定量槪血色素比 率時’必齡财色素錄化血色诚度，纽是說在測 量訊號值可能會有十倍以上的差距，若錢量測方法之偵測範圍 無法^效包含須檢種血色素及糖化血色素濃度之咖時，則最 終計算所得糖化血色素比率受影響並造成量測結果的不準確。至 於本創作不但僅針對私色诚度進行量測；而且僅使用相同檢 測方式壯色诚度進行制。最重要岐所制私色素濃度 範圍至多僅有十倍的差異，由之前習得檢測方式可知量測結果的 M383733 不準確來自上子，縣創作射__彡_聽果的干 擾因素’並有可能將以免疫分析定餘化血色素比補確性提 至與目前實驗室採取之標準檢測方法_之規格。 【實施方式】 有關本創作之詳細綱及技_容，現在配合圖式說明如下： 參一、糖化血色素比率測量裝置 請參閱「第二圖」，係本創作之糖化血色素比率測量裝置的主要 組件示意圖，其具體内容如下： 1. 一反應管4 ’可容納反應物及反應試劑於其内作用，反應試劑包 含用於破壞紅血球的溶血素、及血色素定量試劑，該血色素定量 試劑可與血色素作用而增強吸光值訊號，及生物專一性磁珠，可 •專一性的與糖化血色素結合，受到磁性槽座5的引導至反應管4 底部。這些反應試劑用於提供定量待測物中總血色素濃度及經藉 助磁力吸收之待測物中糖化血色素後而定量待測物中扣除糖化血 色素後之濃度。 2· —有執行量測及免疫作用的反應管4’其定義為具可裝盛反應溶 液且適用於吸光度測量的反應管4。根據Beer-Lambert Law定義， 吸光值Absorbance(OD) = E X C X L，吸光度等於待測物的消光 12 M383733 系數、光路徑及樣品漢度三者相乘，於是反應管4内部的直徑（光 徑為1 cm)，也就是說當反應管4置於反應管槽7中執行测量時的 光:徑是水平且穩固的，如此一來樣品的吸光度量測值才會與樣 品濃度成正比，並隨樣品濃度而增加。 _ 3·位於反應倾7絲具有—雖_5，其定義是可提供磁力使 得反應管4内的磁性物質，這裡是指糖化血色素-抗糖化血色素抗 癱體-生物專-磁珠複合物’在磁力作用下可彙集至反應管4底部， 經作用後反應管4内所懸浮便是未糖化之血色素，及置放反應管4 之反應管槽7側邊兩端具有一量測視窗6，用以讀取企色素濃度量 測值。 二、本創作配合糖化A色素收集裝置之生物專一性磁珠 請參閱「第一圖」，係本創作所導入抗糖化企色素抗體1以共 價鍵3結合之生物專—性磁珠之示意圖，在磁性槽座5搭配下可 收集待測物中糖化i色素4中抗糖化血色素抗體丨係指能辨識 且結合糖化血色素生物性分子，包括：單株或多株型式之抗體。 至於生物專-性磁珠是指以氧化鐵等材質做為核心（core)，外圍 再以石夕為外财裹且含有可倾結的具官能基修飾的磁性微粒 2例如.COOH、HSH等，經修飾並可朝磁場產生方位聚集 之金屬微粒。最後，_化學反應將二者在搭配合適的鍵結試劑 13 M383733 (視可供鍵結的官能基做為挑選依據）以共價鍵3連接成一複合分 子’使其分別具有感測與彙集雙重功能。 二、操作流程及所搭配之計算方程式 用於本創作之糖化血色素比率測量裝置，包括：反應管槽 7反應14及其所含之内容物、磁性槽座5，所需依循之量測 程序。其流程與執行靡的規齡1^加人制樣品（及反應物， 此為全血）’胁反絲4 _域試航分混合，使紅血球被分 轉出血色素蛋白’此時反應試劑另—成分：血色素定量試劑與 血色素蛋白作用，使之可於特定波長下執行血色素濃度量測由 反應管槽7側邊之量測視窗6讀取獲得一讀值以）。 之後以共價鍵3結合抗糖化血色素抗體丨及具官能基修飾的 磁性微粒2與反應管4 _化血色素分子作祕，在藉助磁性槽 座5所提供之磁力作用下，將待測樣品中糖化血色素連同生物專 -性磁珠-併彙集至底部，此時再次於特定波長下執行也色素濃 度量測，由反應管槽7側邊之量測視窗6讀取獲得一讀值⑻。本 創作之糖化血色素比率測量裝置中更包含一計算糖化企色素比 率之裝置，其可量測上述之A#B值後，已知也色素濃度桿 準轉將吸紐換算成血色素濃度，再代人搭配上賴作程序之 計算方程式(Hb Ale % = (A-B)/A_) ’便可計算出糖化血色素 比率。 以上所揭，僅為本創作的較佳實施方式而已，並非用 以限定本創作實施例的範圍，本技術領域内的—般技術人 M383733 員根據本創作所作的均等變化，以及本領域内技術人員熟 知的改變，仍應屬本創作涵蓋的範圍。 【圖式簡單說明】 第一圖本創作使用之生物專一^f生磁珠示意圖 第二圖本創作之裝置示意圖 鲁【主要元件符號說明】 1抗糖化血色素抗體 4 反應管 2具官能基修飾的磁性微粒5 磁性槽座 3共價鍵 6 量測視窗 7 反應管槽 15Hb Ale % = (A-B)/a^i〇〇 The glycated hemoglobin ratio can be obtained. Quantitative glycosylated hemoglobin creation with current immunosuppression technology is implemented in the implementation of _ _ _ _ _ _ _ Ben 9 M383733 1. Traditional mining immunoassay technology to quantify the glycated hemoglobin ratio of the device, its in the targeted measurement of the object 'includes: total Hemoglobin and glycosylated pigments, some of which are used to measure their specific antibodies; or for their use, for example: using anti-glycated hemoglobin antibodies, using antibodies for detection, and then for another The headings use their non-immune means, such as optical analysis of absorbance measurements at specific wavelengths. Whether it is used or only used - using antibodies as a diligent device, the application of immunoassay technology to measure the glycated hemoglobin ratio is a two-point measurement analysis mode. It is also possible to calculate the true glycated hemoglobin ratio of each individual due to the final inclusion of the above two standard measurements (due to the difference in the hemoglobin content of each subject). Therefore, when using the immunoassay technology to quantify the glycated hemoglobin ratio, it is easy to produce large variations due to the integration of the two types or the use of the same type but the integration of two different detection tools. , which led to the development of quantitative glycosylated hemoglobin ratio products using immunoassay technology. • Obstructed i in this creation, by guiding bio-specific (the magnetic bead becomes a marker detection device, under the magnetic slot, The characteristics of the glycated hemoglobin molecules in the reaction tube are collected into the near magnetic housing at the bottom of the reaction tube. At this time, when the optical colorimetric device is used for the measurement, the detected hemoglobin concentration is the hemoglobin concentration after deducting the glycated hemoglobin. On the other hand, the optical hemoglobin is used to perform the measurement when the glycosylated hemoglobin antibody-biospecific magnetic bead complex is not added, and the total hemoglobin concentration is calculated by the equation with the known concentration. Calculate the equation to obtain the ratio of the glycated jk pigment ratio in the test object. If H is in the same-test method, it will be excluded. The quarantine detection technology uses two types of measurement, the two types of saccharification, and the use of two 10 M383733. The two types of measurement are divided into two types, which are caused by the county entanglement _ detection method, which leads to excessive variation of the final measurement results. ·Because of the adoption of immunoassay technology, it is more important than the traditional quantitative glycosylated hemoglobin to read hemoglobin. However, it is necessary to remove the sample from the sample under the cross-reaction process. Performing the glycosylation hemoglobin concentration surface is inevitable. Said (4). In this way, it will inevitably lead to an increase in manufacturing costs and a good county. She is selected as an optical cuvette that can be used routinely. The special design of the magnetic housing can be used to separate the hemoglobin molecules. It should be able to know the ratio of glycated hemoglobin in the test sample, and estimate the cost of the whole reading material is also far from the commodity of the servant. 3. Known-like people The normal value of glucosinin is about 4%, and the glycated hemoglobin value of diabetic patients is as much as possible (4) in _τ. However, when the ratio of sputum hemoglobin is quantified, Degree, New Zealand means that there may be more than ten times the difference in the measured signal value. If the detection range of the money measurement method cannot be used to include the blood sugar and glycated hemoglobin concentration, the final calculated glycated hemoglobin ratio Affected and caused inaccurate measurement results. As for this creation, not only the measurement of private color integrity; but also the use of the same detection method to achieve color and integrity. The most important thing is that the range of private pigment concentration is at most Ten times the difference, from the previous acquisition detection method, the measurement result M383733 is inaccurate from the son, the county creates a __彡_the fruit's interference factor' and may use the immunoassay to determine the hemoglobin ratio The specifications of the standard test method adopted by the current laboratory. [Embodiment] The detailed outline and technical information of this creation are as follows: Refer to the second figure for the measurement of glycosylated hemoglobin ratio. The main components of the glycated hemoglobin ratio measuring device of the present invention are as follows: 1. A reaction tube 4' can accommodate the reaction And a reaction reagent comprising a hemolysin for destroying red blood cells and a hemoglobin quantifying reagent, the hemoglobin quantifying agent can enhance the absorbance signal and the biospecific magnetic bead with hemoglobin, and can be specific In combination with the glycated hemoglobin, it is guided by the magnetic housing 5 to the bottom of the reaction tube 4. These reagents are used to quantify the total hemoglobin concentration in the test substance and the glycated hemoglobin in the test object by magnetic absorption to quantify the concentration of the glycated hemoglobin after subtraction of the test substance. 2· A reaction tube 4' having a measurement and an immunological function is defined as a reaction tube 4 having a reaction solution and suitable for absorbance measurement. According to the Beer-Lambert Law definition, the absorbance Absorbance (OD) = EXCXL, the absorbance is equal to the extinction of the object to be tested 12 M383733 coefficient, the light path and the sample Han multiplied, so the diameter of the inside of the reaction tube 4 (light path is 1 Cm), that is to say when the reaction tube 4 is placed in the reaction tube tank 7 to perform the measurement of the light: the diameter is horizontal and stable, so that the absorbance measurement value of the sample is proportional to the sample concentration, and with the sample Increase in concentration. _ 3· Located in the reaction, the 7-filament has - although _5, which is defined as a magnetic substance that can provide magnetic force in the reaction tube 4, here refers to a glycated hemoglobin-anti-glycated hemoglobin anti-steroidal-biospecific-magnetic bead complex' Under the action of magnetic force, it can be collected to the bottom of the reaction tube 4. After the action, the suspension in the reaction tube 4 is unsweetened hemoglobin, and the measuring tube 4 is placed at both ends of the reaction tube 4 to have a measuring window 6 at both ends. It is used to read the measured value of the pigment concentration. 2. The bio-specific magnetic beads of this creation combined with the glycation A pigment collection device, please refer to the "first map", which is a schematic diagram of the bio-specific magnetic beads in which the anti-glycation antibody 1 is combined with the covalent bond 3. The anti-glycated hemoglobin antibody in the saccharified i-glycan 4 can be collected and combined with the glycated hemoglobin biological molecules, including single-type or multi-type antibodies. As for the bio-specific magnetic beads, a material such as iron oxide is used as a core, and the outer periphery is wrapped with a stone-like outer layer and contains a pegable magnetic particle having a functional group modification such as COOH, HSH, etc. Metal particles that have been modified and can be aggregated toward the magnetic field. Finally, the _ chemical reaction combines the two with a suitable bonding reagent 13 M383733 (depending on the functional group available for bonding) and a covalent bond 3 to form a composite molecule Features. Second, the operation process and the calculation equations used in this creation of the glycated hemoglobin ratio measuring device, including: reaction tube tank 7 reaction 14 and its contents, magnetic housing 5, the required measurement procedure. The process and the implementation of the 靡 规 ^ 加 加 加 加 加 加 加 加 加 加 加 加 胁 胁 胁 胁 胁 胁 胁 胁 胁 胁 胁 胁 胁 胁 胁 胁 胁 胁 胁 胁 胁 胁 胁 胁 胁 胁 胁 胁 胁 胁 胁 胁 胁 胁Ingredients: The hemoglobin quantification reagent interacts with hemoglobin to perform hemoglobin concentration measurement at a specific wavelength and is read by the measurement window 6 on the side of the reaction tube groove 7 to obtain a read value. Then, the covalent bond 3 is combined with the anti-glycated hemoglobin antibody 丨 and the magnetic particle 2 modified with the functional group to react with the reaction tube 4 _ hemoglobin molecule, and the sample to be tested is subjected to the magnetic force provided by the magnetic housing 5 The glycated hemoglobin is combined with the biospecific magnetic beads - and collected to the bottom. At this time, the pigment concentration measurement is performed again at a specific wavelength, and a reading value (8) is obtained by reading from the measurement window 6 on the side of the reaction tube groove 7. The saccharified hemoglobin ratio measuring device of the present invention further comprises a device for calculating the ratio of saccharification and pigmentation. After measuring the A#B value, it is known that the pigment concentration rod is converted into a hemoglobin concentration, and then replaced by a person. The glycated hemoglobin ratio can be calculated by using the calculation equation of the procedure (Hb Ale % = (AB)/A_). The above description is only for the preferred embodiment of the present invention, and is not intended to limit the scope of the present creative embodiment. The average change made by the general practitioner M383733 in the art according to the present invention, and the technology in the field. Changes familiar to the person should still be covered by this creation. [Simple description of the diagram] The first picture of the bio-specific ^f magnetic beads schematic diagram of the second picture of the creation of the device schematic diagram [main components symbol description] 1 anti-glycated hemoglobin antibody 4 reaction tube 2 functional group modified Magnetic Particles 5 Magnetic Slots 3 Covalent Keys 6 Measuring Window 7 Reaction Tube Slots 15