利用即時定量聚合酶鏈鎖反應技術執行快速 ABO 血型分子檢驗鑑定

Rapid TaqMan qPCR assay for ABO Blood Typing

ABO基因分型常應用於多種情況，包括個人識別，疾病檢測和輸血。本研究利用水解探針搭配快速即時定量聚合酶鏈鎖反應技術，針對ABO血型基因核苷酸261（rs8176719）及796（rs8176746）位點，進行血液及口腔黏膜檢體之ABO血型檢驗，檢驗結果可進一步區分成AA, AO, BB, BO, AB及OO六種基因型。執行84例檢測樣本的基因型檢驗，確認共有AA型1位、AO型17位、BB型4位、BO型21位、OO型34位及AB型7位；分子檢驗與血清學檢驗方法兩者結果並無差異。本研究作業流程於一小時內即可得到正確檢驗結果，所使用之DNA濃度僅需10 ng。實驗結果證實，本研究所提供之分子檢驗方法，可做為輔助傳統血清學檢驗鑑定ABO血型的良好方法。利用可輕易取得含有細胞核之檢體，如口腔黏膜細胞等來鑑定ABO血型，可減少對新生兒採血量及次數。

ABO genotyping is commonly used in several situations, including personal identification, disease detection and blood transfusion. By rapid identifying two SNP nucleotides (rs8176719 and rs8176746), eighty-four blood and oral mucosa samples were determined to be AA (n=1), AO (n=17), BB (n=4), BO (n =21), OO (n=34), and AB (n=7). There is no discrepancy between phenotypes (determined by serological assay) and the genotypes (determined by qPCR assay). This method enabled analyses to be completed within one hour. The DNA concentration required for the experiment was 10 ng per reaction in each SNP site. Easy access oral mucosal cells instead of blood phlebotomy in order to identify ABO blood genotype. This rapid method, with good accuracy, provides a supplement to routine serological ABO typing.