利用奈米金增強電化學阻抗分析探針發展快速檢測抗穆勒氏荷爾蒙平台

Abstract

抗穆勒氏荷爾蒙(Anti-Mullerian hormone)，簡稱AMH，又名為穆勒氏抑制物質(Mullerian inhibiting substance, MIS)，是轉化生長因子-家族(TGF-family)的一員。在婦女妊娠期(gestation) 7週時，可於男性胎兒血液中發現AMH，此時的AMH是由塞托利氏細胞(Sertoli cell)所分泌，分泌的AMH會與AMH receptor結合並抑制穆勒氏管的演化，導致無法發育形成子宮。在女性胎兒，AMH可於妊娠期36週時被偵測到，此時的AMH是被前有腔濾泡(preantral)的顆粒性細胞(granulosa cell)所分泌。另外，婦女體內的AMH濃度可顯示其卵巢儲備量，因此於生殖醫學鑑定方面，AMH已被廣泛應用於評估是否適合進行人工催卵，但目前之檢測方式的穩定性與檢測時間均有待加強。又電化學阻抗譜儀，簡稱EIS，其利用交流信號提供用於檢測的少量生物分子的信息。在我們的實驗室中，我們建立了一個簡單，快速，可靠，靈敏的生物感測檢測平台，以量化大腸桿菌71型(Enterovirus 71, EV71)，血紅蛋白(Hemoglobin)與血紅素結合蛋白(Haptoglobin)的濃度，並在本研究中，我們運用已有之技術平台創建了一個具AMH專一性的探針以進行AMH快速檢測及定量，從樣品浸泡以及電化學頻譜儀偵測的時間僅需約18分鐘，並可與傳統的酵素免疫分析法ELISA進行比較。這會是一個快速AMH檢測平台並可用來量化婦女血清中的AMH。

Anti-Mullerian hormone (AMH), also named Mullerian inhibiting substance (MIS), is a member of the transforming growth factor-beta (TGF-) family. This hormone regulates regression of Mullerian ducts in male fetuses and the hormone is produced by fetal Sertoli cells of the fetal testis at 7 weeks of gestation. In females, AMH is secreted by the granulosa cells of the preantral and small antral follicles; it can be detected in human fetal ovaries as early as 36 weeks of gestation. AMH has been found to be an excellent marker of ovarian reserve in women. That is, the expression level of AMH in women had been used to evaluate the feasibility of artificial ovulation. Moreover, the stability and detection time of traditional method should be improved. It is known that the AC signal of electrochemical impedance spectroscopy (EIS) provides more information for detecting the small amount bio-molecules. In our laboratory, we had established a simple, fast, reliable and sensitive biosensor platform to quantify concentration of Ev71, Hemoglobin and Haptoglobin. In this study we have created a specific probe for rapid detecting and quantification of AMH. At the same time, the AMH binding could be detected and quantified within 18 min. This is a new platform to quantify AMH in the serum of women to our knowledge. The EIS methods were compared with ELISA in this study, too.