2023-08-012024-05-15https://scholars.lib.ntu.edu.tw/handle/123456789/665997隨著精準醫療觀念興起，「精準輸血(precision blood transfusion)」的重要性也逐年受到重視。個人化的醫療與檢驗的觀念逐年普及於醫療端，也漸擴及於病人及一般大眾，也因此去中心化的定點照護檢驗(point-of-care testing、POCT)形成未來的趨勢。 GP.Mur是台灣最具臨床重要性的血型。然而各種不同、複雜的GP.血型難以用傳統血清學明確區別；中心化的高通量定序要精準鑑別也尚有其困難度；同時基因定序也難以應用至一般檢驗程序。 本研發計畫所設計構想GP.Mur血型及基因合子型(zygosity)的POCT流程，包括以recombinase polymerase amplification (RPA)進行等溫核酸擴增(isothermal nucleic acid amplification)、以Exo probe搭配RPA反應的螢光結果判讀(florescence readout)、以Nfo probe搭配側向流體免疫分析的結果判讀(lateral flow readout)，以及CRISPR-based nucleic acid detection。 初步成果顯示，計畫中區別GYPA、GYPB、和GP.Mur基因及基因合子型的部分關鍵設計是成功、可行的（詳見”研究方法”）。 本研發計畫將逐步執行並整合下列四大項：(1)各項設計構想的持續研發測試、(2)實體樣本測試、(3) POCT系統整合、(4)靈敏度(sensitivity)與特異度(specificity)的調整及驗證。 簡易、快速的GP.Mur血型定點照護分子檢驗套組除了可於醫療端使用，以滿足長期、需重覆輸血病人的需求外，更在近年醫療及檢驗個人化觀念的普及與趨勢下，可推展至個人的居家檢測(home self-testing kit)。 方便、快速的鑑別GP.Mur血型的實踐對台灣推展「精準輸血」是重要的一步。 With the rise of the concept of precision medicine, “precision blood transfusion” has attracted more and more attention in recent years. Personalized medical treatment and testing has been well recognized in the medical field and the concept has gradually spread to patients and the general public. This makes the decentralized point-of-care testing (POCT) a future trend. GP.Mur is the most clinically important blood type in Taiwan. Anti-Mia induced by GP.Mur is the most common alloantibody in Taiwan and is a clinically significant antibody that needs attention. Nevertheless, due to the complex serology of different GP. blood types and the rarity of specific antibodies for specific GP. antigens, it is difficult to use traditional serological test to identify specific GP. blood type accurately. And due to the high degree of homology among the gene sequences of different GP. blood types, molecular typing is also difficult to accurately distinguish GP. blood types. This R&D project aims to establish a POCT kit for identifying GP.Mur blood type and determining gene zygosity. The designed POCT processes include isothermal nucleic acid amplification using recombinase polymerase amplification (RPA), Exo probe in RPA reaction with fluorescence readout and Nfo probe with lateral flow readout to identify GYPB and GP.Mur genes, and CRISPR-based nucleic acid detection. Our preliminary results demonstrate that the key designs in RPA reactions and the Exo and Nfo probes to distinguish GYPA, GYPB, and GP.Mur genes and determine the gene zygosities are successful. (Detailed in “Research Methods”) This project will implement and integrate the following four major researches steps by steps: 1. Continuing developing and testing the designs and schemes proposed in the project, 2. Testing the true GP.Mur-blood type samples with different zygosities, 3. Integration of the POCT system, 4. Adjustment and verification of the sensitivity and specificity of the POCT system. Simple and fast POCT kit for GP.Mur-blood group typing and zygosity determination can be used in laboratory testing to meet the needs of patients who need repeated blood transfusions, and with the popularization and the trend of personalization of medical care and testing in recent years, it can be extended to personal use for home-based diagnostics. The practice of accurate and convenient determination of GP.Mur blood group is an important step to push forward "precision blood transfusion" in Taiwan.米田堡血型;定點照護;分子檢驗;精準醫療;基因型;blood groups; Miltenberger; point-of-care testing; precision medicine; genotyping