本文選題：單壁碳納米管　切入點：熒光標(biāo)記的核酸探針　出處：《生物技術(shù)通報》2017年12期 　論文類型：期刊論文

【摘要】：基于熒光能量共振轉(zhuǎn)移(Fluorescence energy resonance transfer,FRET)原理利用單壁碳納米管(Single-walled carbon nanotubes,SWCNTs)構(gòu)建一種操作簡單、成本低、靈敏度高和選擇性強的納米生物傳感器快速檢測含有tst基因金黃色葡萄球菌。將tst基因互補序列設(shè)計為探針,利用單壁碳納米管聯(lián)合羧基熒光素(FAM)標(biāo)記的DNA分子探針(FAM-P)構(gòu)建出FAM-P/SWCNTs復(fù)合檢測體系。考察該體系對靶標(biāo)序列和靶標(biāo)菌的靈敏度和特異性。結(jié)果表明,該生物傳感器對靶標(biāo)序列的檢測下限低至10 nmol/L,并在10-100 nmol/L范圍內(nèi)具有較好的線性關(guān)系(R2=0.994),對靶標(biāo)菌的檢測下限低至102,且在102-107CFU/m L范圍內(nèi)均呈現(xiàn)良好的線性關(guān)系(R2=0.999)。此外,在錯配實驗和對非靶標(biāo)菌檢測時,該傳感器呈現(xiàn)了較高的特異性。本研究構(gòu)建的FAM-P/SWCNTs體系能快速檢測含tst基因金黃色葡萄球菌,并具有操作簡單、靈敏度高、特異性強等優(yōu)點,為食品檢測、臨床診斷等領(lǐng)域開辟了新途徑。
[Abstract]:Based on the principle of fluorescence energy resonance transfer (fret), a novel single-walled carbon nanotube SWCNTs was constructed with simple operation and low cost. A sensitive and selective nanosensor for rapid detection of staphylococcus aureus containing tst gene was developed. The complementary sequence of tst gene was designed as a probe. The FAM-P/SWCNTs complex detection system was constructed by using single-walled carbon nanotubes (SWNTs) and carboxyl fluorescein (FAM) labeled DNA probe FAM-P.The sensitivity and specificity of the system to the target sequence and the target bacteria were investigated. The detection limit of the target sequence by the biosensor is as low as 10 nmol / L, and the linear relationship is good in the range of 10-100 nmol/L. The detection limit for the target bacteria is as low as 102, and the linear relationship is good in the range of 102-107CFU/m L. The FAM-P/SWCNTs system constructed in this study can detect staphylococcus aureus containing tst gene quickly and has the advantages of simple operation, high sensitivity, strong specificity and so on. For food detection, clinical diagnosis and other fields opened up a new way.
【作者單位】： 中國醫(yī)學(xué)科學(xué)院北京協(xié)和醫(yī)學(xué)院醫(yī)學(xué)生物學(xué)研究所中心實驗室;長沙醫(yī)學(xué)院基礎(chǔ)醫(yī)學(xué)院應(yīng)用生物學(xué)教研室;
【基金】：高等學(xué)校博士學(xué)科點專項科研基金資助項目(20111106120056)
【分類號】：R446.5

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