評估染劑轉換對於一市售雞隻微陣列核酸晶片之效能影響

核酸微陣列晶片分析技術能夠同時檢測大量基因的表現量差異，甚至能觀察到未知的調控路徑，是一種在探討基因變化相當實用的研究工具。然而此技術會因染劑與不同基因親和性或是染劑自身穩定度等因素，而影響晶片結果分析上的特異性及敏感度。隨著雞隻的全基因序列被全部解析，目前雖已有商品化的雞隻基因微陣列晶片，仍需要審慎評估其效能及使用上所必須注意的篩選標準。本實驗利用染劑轉換的試驗設計，評估一市售雞隻微陣列晶片其可能存在之染劑偏差效應及篩選標準的關係。結果發現的確存在染劑偏差的效應，因此該晶片數據需經適當的篩選進行進一步分析，當差異倍數判斷值設定在1.5倍時，染劑的偏差效應尚不足以影響基因改變量差異的判讀結果，因此建議後績研究者可使用此倍率做為判斷值；同時若未經過訊號強度（raw值）篩選的數據，即使將判斷標準提升至差異2倍的較高標準，也發現其迴歸線R^2值降低且整齊度不佳，結果可信賴度不佳，說明了以訊號強度做為篩選條件之一的重要性。本評估結果可供其他研究者日後進行本型晶片研究分析時的參考指標。

關鍵字

微陣列晶片；染劑偏差；染劑轉換；基因表現量；訊號強度篩選

並列摘要

Microarray has been considered as a useful tool for the studies of gene expression level changes. It is capable of detecting target cell gene expression level difference in a global view and even offers information of undiscovered regulation pathways. Meanwhile, the sensitivity and specificity of this technique has been questioned due to the stability of the dye and its affinity to genes. With the whole genome has been sequenced, commercial chicken whole genome microarray is available. However, the efficacy and the qualitative standard should be evaluated prior to the usage of the microarray. In the current study, an evaluation to dye-bias and qualitative standard was con- ducted by a dye-swap experiment. The results showed the existence of the dye-bias, and therefore a proper qualitative standard is needed for further study. It was also found that when the cut-off ratio of expression difference was set as 1.5-fold, the dye-bias would not be strong enough to interfere with the analysis results. Furthermore, even the cut-off ratio was lifted to 2.0-fold, data without being signal strength screened showed a low reliability. This result indicates the need of conducting signal strength screening during microarray analysis. This study provides useful information for further studies using commercial chicken microarray.

並列關鍵字

microarray ； dye bias ； dye swap ； gone expression level ； signal strength screening

國際替代計量

評估染劑轉換對於一市售雞隻微陣列核酸晶片之效能影響

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