檢測弱抗原血型之微流體凝膠管柱晶片製備

Fabrication of Microfluidic – based Gel Column for the Determination of Weak Blood Group

黃至堅(Chih-Chien Huang)

指導教授 ： 謝明發

中原大學/工學院/生物醫學工程研究所/碩士(2012年)

https://doi.org/10.6840/cycu201201011

摘要

人類血型中除了常見的ABO型外還有Rh血型；而 – Del – 血型(D elution)為RhD血型中紅血球表面抗原量最少的血型，無法藉由一般血型篩檢快速判別Del 血型而造成輸血危險，如新生兒溶血症、血栓或全身性溶血症等不良反應。而傳統篩檢 Del 血型時，檢驗過程中紅血球與抗體反應時間過於費時，且需花費大量的檢體與試劑，臨床上期望能縮小檢體與試劑的用量、降低檢測過程中所耗費之時間且須準確的檢驗出弱抗原血型。本研究是結合凝膠管柱及微流道晶片製作成一可用於血液凝集測試之微量管柱，其微流道長10微米、寬1微米與高1微米；將層析凝膠注入一微流道中，並在微流道中注入血液與各種抗體進行血液凝集分層，所用之分層透析凝膠的體積為3 毫克、血液用量25微升與檢測液體的用量如抗體使用量25微升比較傳統檢測法所使用的樣品量300至500微升與常規檢測法的使用量皆有大幅降低，所以耗費的成本也將大幅度的降低，且本研究之微管柱晶片不單用於弱抗原血型之凝集使用，也可用於一般ABO血型常規檢測中使用，並且利用簡單的方式製做出微凝膠管柱晶片來進行血液檢測，且進行單次檢測費時5至10分鐘；本研究進行了多組檢體的實驗並與臨床上所使用之傳統檢測法進行比較，其結果皆為相同；其可望開發出一檢體與檢測液體使用量少於傳統檢測法並減少檢測時間且能精確檢驗弱抗原反應之微流道晶片。

關鍵字

分層透析膠 ； 血液檢測 ； 弱抗原血型 ； 微管柱

並列摘要

Beside the ABO blood types, there’s another one which’s called as Rh blood type. The false negatives of red blood cells (RBCs) with Rh (Del) cannot be quickly determined by the general blood center using the method Ortho BioVue System Microtyping card. The result in subsequent disease such as hemolytic disease of newborn、thrombus and hemolytic infants. The traditional method of identifying Del, namely the adsorption and elution methods are time-consuming, require large quantities of samples and reagents hence, it raises the cost of pre-transfusion testing. It’s expected to reduce the sample、agents usages 、require time in the clinic trials and hope to detect weak blood group precisely. This study therefore aims to fabricate a microfluidic-based gel column to address the problem above by combining sephadex-gel with the microfluidic chip (PDMS chip). The length is 10 mm, width is 1 mm and the height is 1 mm. This column was used as the blood agglutination test. The blood is combined with a variety of antibodies then injected into the microfluidic chip and coagulate into the microflow channel. The new material requires only approximately 25 microliters each of the blood sample and reagent and exactly 0.003 g of sephadex gel to produce one column. These values are much lower than what is required by currently used procedure for blood agglutination test. Therefore, the newly fabricated material relatively reduces cost for blood agglutination test mainly because it uses only very small quantities of blood sample hence the reagents to react with it and cost 5 to 10 minutes for each test. This study has done a lot of experiments for sample and compare with the traditional method which’s used in clinic, and came the same results Furthermore, the new device can quickly be used for blood agglutination test to determine blood type with weak antigen. In addition, it can also be used for the detection of the general ABO blood groups.

並列關鍵字

Blood test ； weak antigens ； Blood groups ； Microfluidic-based Gel Column ； Sephadex-gel

參考文獻

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