本文選題：牛流行熱 + 病毒檢測�。� 參考：《華南農(nóng)業(yè)大學(xué)學(xué)報(bào)》2017年03期

【摘要】：【目的】建立一種能夠快速、簡便、可視化地檢測牛流行熱病毒的分子生物學(xué)方法。【方法】根據(jù)牛流行熱病毒G蛋白基因的6個(gè)保守區(qū)域設(shè)計(jì)2對引物,建立牛流行熱病毒可視化逆轉(zhuǎn)錄環(huán)介導(dǎo)等溫?cái)U(kuò)增(Reverse transcription loop-mediated isothermal amplification,RT-LAMP)檢測技術(shù),優(yōu)化RT-LAMP的反應(yīng)條件,將其與PCR方法進(jìn)行比較�！窘Y(jié)果】當(dāng)Mg2+濃度為3 mmol·L~(-1)、甜菜堿濃度為0.4 mol·L~(-1)、d NTPs mix濃度為1.2μmol·L~(-1)、內(nèi)外引物濃度比例為8∶1、反應(yīng)溫度為63℃時(shí),反應(yīng)梯形條帶最明顯,在反應(yīng)40 min后可以觀察到明顯的梯形條帶。建立的RT-LAMP檢測方法特異性好,只對牛流行熱病毒進(jìn)行擴(kuò)增;靈敏度比普通PCR高10倍�！窘Y(jié)論】該方法操作簡便,特異性強(qiáng),結(jié)果判讀方便,可用于牛流行熱的快速檢測。
[Abstract]:[objective] to establish a molecular biological method for rapid, simple and visual detection of bovine epidemic fever virus. [methods] two pairs of primers were designed according to 6 conserved regions of bovine prevalent fever virus G protein gene. In order to optimize the reaction conditions of RT-LAMP, a visualized reverse transcription loop-mediated isothermal amplification (RT-LAMPMPP) technique was established for the detection of bovine epidemic fever virus (BVV) by reverse transcription loop-mediated isothermal amplification. [results] when the concentration of Mg2 was 3 mmol / L ~ (-1), the concentration of betaine was 0.4 mol / L ~ (-1), the concentration of mix was 1.2 渭 mol / L ~ (-1), the ratio of internal and external primer was 8: 1 and the reaction temperature was 63 鈩，

本文編號：2018548