癌症基因突變之生物檢測器研發與探討

近年來罹患癌症比率越來越高而且每個癌症起因很複雜，近三十年以來都以螢光原位雜合技術及鹼基定序法進行檢測基因，但耗時久、成本高等原因，促使需要研發有更快、更省成本的方法偵測突變點。我們的目標是要研發可有效偵測癌症相關基因突變的生物感測器，而我們設計三種突變型式的探針:刪除、插入、置換及野生型探針。將探針與目標基因片段進行雜合反應，確認偵測核苷酸配對數目的不同，而C-V曲線也會有差異出現，因此可以更快找出該癌症的基因突變區域並且採取適當的臨床用藥策略。我們實驗在pH 7緩衝液中進行，利用各種突變型式之探針進行雜合，並經由C-V曲線的位移差異性來確定其基因突變。從這些感測實驗的結果可知目標基因片段與各種探針雜和後有錯誤配對的存在，其C-V曲線皆有差異性。實驗結果得知此半導體DNA生物感測器可非常迅速偵測不同型態之基因鹼基突變。未來仍需持續提高生物感測器的準確性及穩定性，期許在臨床癌症相關DNA的突變偵測能有很大之幫助。

關鍵字

螢光原位雜合技術；生物感測器；探針；突變偵測

並列摘要

There had been a significant increase in the cancer incidence, but the underlying causes for each type of cancer remain complex. Gene’s mutations detection often was used in the past three decades that used of fluorescence in situ hybridization technique and nucleotide sequencing has been the predominant method. However, these methods are time-consuming and expensive. For these reasons, a more rapid and cost-effective method for detecting gene mutations needs to be developed. Therefore, the aim of our study is to develop a biosensor that can be used to effectively detect gene mutations that may be associated with cancer. Firstly, a fragment of the target gene is hybridized to each of the wild-type and mutational probes. The differences in the number of pairing nucleotides between each hybridization are then determined, and this can be used to generate different C-V curves. Thus, the certain region of cancer gene that contains the mutation can be rapidly determined, and provided the appropriate clinical treatment strategies can be based on this information. We conducted experiments in the pH 7 buffer, using various types of mutations probe is hybridized to fragment of the target gene; and then observation C-V curves of shift differences to determine gene mutations. From these sensing experiments show that fragment of the target gene is hybridized to each of mutational probes presence mismatch pairs, cause have differences in C-V curves. The results show that the semiconductor DNA biosensor can detect different types of gene mutations rapidly. In the future, the accuracy and stability of the biosensor still needs improvement, and expect can be a great help for detection of cancer-associated DNA mutations in clinical.

並列關鍵字

Fluorescence in situ hybridization (FISH) ； biosensors ； probes ； mutation detection

參考文獻

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癌症基因突變之生物檢測器研發與探討

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