傳統的戴奧辛檢測大都藉由如高解析度質譜儀等化學方法進行，其分析雖相當準確但相對成本非常昂貴且耗時，因此，若欲全面瞭解國內環境介質是否受戴奧辛污染，應建立快速且高敏感度的生物檢測系統。戴奧辛的生物毒性，主要是經由芳香族碳氫化合物受器（Aryl hydrocarbon receptor, AhR）所誘發。我們利用轉殖技術，建構一個含有dioxin responsive element（DRE）啟動子（promoter）之冷光酶基因（luciferase gene），並將含此基因之質體殖入人類的肝臟細胞中，經藥物篩選後，挑選出能夠以戴奧辛誘發冷光酶大量表現之穩定細胞株，當戴奧辛或類戴奧辛物質與此細胞內之AhR結合後，即會誘發冷光酶基因表現，因此，藉由偵測冷光酶的活性強度，可間接計算這些戴奧辛或類戴奧辛物質之總毒性當量。在此研究中，我們對胎盤檢體進行戴奧辛含量的生物及化學檢測，胎盤檢體的收集以台灣中部地區健康的生產婦女為主要對象，檢體選取標準包括：願意提供胎盤、完成問卷、在中部地區居住至少五年、無多氯聯苯（PCB）暴露病史，隨機選取32位產婦胎盤；經兩種不同固相萃取流程後，利用此細胞株進行生物檢測，分析胎盤檢體內戴奧辛含量之總毒性當量，分析結果顯示，此生物檢測方法偵測極限值為 0.19 pg-CALUX-TEQ/g lipid [萃取流程1：平均測值＝10.7 pg-CALUX-TEQ/g lipid, 標準偏差＝7.80 pg-CALUX-TEQ/g lipid；萃取流程2：平均測值＝36.9 pg-CALUX-TEQ/g lipid, 標準偏差＝11.9 pg-CALUX-TEQ/g lipid]；另外，在化學分析方面，相同的胎盤檢體，經離心、分離、去極性、濃縮等前處理步驟後，將含有戴奧辛及多氯聯苯之濃縮萃取液，以氣相層析高解析度質譜儀進行定性及定量之分析；由生物及化學檢測所得之數據，經統計分析結果顯示，兩者之間並無相關性（p ﹥0.5），因此，欲獲得更完整的資訊，建議進一步將樣本數提高。

Traditional chemical analysis, i.e., high-resolution gas chromatography and mass spectrometry, has been wildly adopted in dioxin measurements for its high sensitivity. However, the chemical analysis is costly and very time-consuming. Therefore, in the meantime, it is practically unlikely to monitor the whole spectrum of dioxin contamination in Taiwan by using the chemical analysis. The objective of this study is to develop a sensitive and high-throughput bioassay, i.e., CALUX, for detection of dioxin contaminations. The dioxin-induced toxicity is mainly through activation of aryl hydrocarbon receptor (AhR). We have cloned a DNA fragment with dioxin responsive elements (DRE) into the promoter region of a luciferase gene. Plasmid carrying the DRE-luciferase gene was transfected into human hepatoma cell lines. After G418 selection, the stably transfected cells with higher inductions of luciferase activity by dioxin were cloned. Upon binding with dioxin/dioxin-like chemicals, the ligand-activated AhR will then activate luciferase expression. Thus, total toxicity equivalency (TEQ) of dioxin compounds can be calculated from the readings of luciferase activity. In this study, we analyzed dioxin levels in human placenta samples by using bioassay (CALUX) and chemical analysis (HRGC/HRMS). The placenta samples were collected from those healthy pregnant women residing in central Taiwan. The recruited human subjects required fulfilling the following criteria, including willing to provide placental specimens, completing the questionnaire, with at least 5-year residence in central Taiwan, and no history of PCB exposure. Thirty-two specimens were randomly selected from the qualified participants. Then, we employed two different solid-phase extraction protocols to extract dioxin/dioxin-like chemicals form placenta samples for bioassay. Our data showed that dioxin detection limit of the bioassay is 0.19 pg-CALUX-TEQ/g lipid [Extraction 1: the average=10.7 pg-CALUX-TEQ/g lipid, standard deviation (SD)=7.80 pg-CALUX-TEQ/g lipid; Extraction 2: the average=36.9 pg-CALUX-TEQ/g lipid, SD=11.9 pg-CALUX-TEQ/g lipid]. For chemical analysis, the same placenta samples were homogenized, centrifuged, separated, depolarized, and concentrated before analysis. The extracts were identified and quantified by using HRGC/HRMS. Statistical analysis showed that there is no significant correlation (p＞0.5) between results from bioassay and chemical analysis. Therefore, it is suggested to increase the sample size for the analyses in order to get more information regarding the application of the CALUX bioassay.

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