比較四種市售反轉錄酶商品在定量BCR-ABL1融合基因中的優劣

Comparison of Four Commercial Reverse Transcriptases for Detection of BCR-ABL1 Fusion gene

定量BCR-ABL1在評估慢性骨髓性白血病的療效，已是一種成熟且被全球實驗室廣為應用的方法。其中，cDNA的品質對於檢測BCR-ABL1 fusion gene的敏感度有很大的影響，進而影響到可測量殘留疾病量（measurable residual disease, MRD）的檢測正確性。本實驗根據ABL1和BCR-ABL1融合基因擴增的成功率，對四種可用於合成cDNA的市售反轉錄酶，分別對BCR-ABL1 fusion gene P210及P190的檢測結果進行評估。根據我們的實驗結果，Invitrogen Super Script™ IV VILO™反轉錄酶試劑盒在本實驗中用於cDNA合成最為理想，其次是由羅氏應用科學提供EvoScript Universal cDNA Kit。 Quantification of BCR-ABL1 to measure the measurable residual disease (MRD) in chronic myeloid leukemia (CML) patients is a well-established method used by clinical laboratories world-wide. The quality of cDNA is critical for high sensitivity in the detection of MRD. Herein, we evaluated four commercially available kits for the synthesis of cDNA. We compared the amplification success rate, and copy numbers of ABL1 and BCR-ABL1 fusion (P210 and P190) transcripts. Our results showed that Invitrogen SuperScript™ IV VILO™ Reverse Transcriptase kit had the best performance, followed by the EvoScript Universal cDNA Kit (Roche Applied Sciences).