目的使用R6G-ddATP作为双脱氧荧光底物建立单碱基末端延伸(SNaPShot)-凝胶荧光法快速检测3种高危型人乳头瘤病毒(high risk human papillomavirus,HR-HPV)(HPV18、HPV33、HPV35)基因型。方法使用HPV质控品作为样本,R6G-ddATP双脱氧荧光...目的使用R6G-ddATP作为双脱氧荧光底物建立单碱基末端延伸(SNaPShot)-凝胶荧光法快速检测3种高危型人乳头瘤病毒(high risk human papillomavirus,HR-HPV)(HPV18、HPV33、HPV35)基因型。方法使用HPV质控品作为样本,R6G-ddATP双脱氧荧光试剂作为底物,首先利用通用引物对HPV进行扩增,得到第一轮扩增产物,经纯化后作为后续SNaPShot反应的模板;然后利用特异性的一步延伸引物进行SNaPShot反应,生成带有R6G荧光标记的DNA延伸产物;产物经过琼脂糖凝胶电泳,在凝胶成像仪下观察电泳结果,通过不同的一步延伸引物对HPV进行分型。每个样本均重复检测3次,并与DNA测序结果进行比较。结果优选的SNaPShot反应的退火温度为55℃;仅需3 h即可对HPV进行分型;在该最适条件下使用R6G-ddATP/SNaPShot-凝胶荧光法检测3种HPV基因型,检测结果与测序结果一致。结论成功建立了3种HR-HPV基因型的微量检测方法——R6G-ddATP/SNaPShot-凝胶荧光法,可用于HPV基因型的快速检测。展开更多
Europium ternary complex of Eu(NNA) 3(NNA=α-naphthylacetic acid) was embedded in silica matrix at different ratios by sol-gel method. The luminescence properties of silica composites were studied by comparing them wi...Europium ternary complex of Eu(NNA) 3(NNA=α-naphthylacetic acid) was embedded in silica matrix at different ratios by sol-gel method. The luminescence properties of silica composites were studied by comparing them with those of corresponding pure complex by means of excitation, emission spectra and lifetimes. The fluorescence lifetime was prolonged when the pure complex was incorporated in silica matrix. The relative fluorescence intensity and fluorescence lifetimes increased simultaneously with the increase of (Eu(NNA) 3.)展开更多
文摘目的使用R6G-ddATP作为双脱氧荧光底物建立单碱基末端延伸(SNaPShot)-凝胶荧光法快速检测3种高危型人乳头瘤病毒(high risk human papillomavirus,HR-HPV)(HPV18、HPV33、HPV35)基因型。方法使用HPV质控品作为样本,R6G-ddATP双脱氧荧光试剂作为底物,首先利用通用引物对HPV进行扩增,得到第一轮扩增产物,经纯化后作为后续SNaPShot反应的模板;然后利用特异性的一步延伸引物进行SNaPShot反应,生成带有R6G荧光标记的DNA延伸产物;产物经过琼脂糖凝胶电泳,在凝胶成像仪下观察电泳结果,通过不同的一步延伸引物对HPV进行分型。每个样本均重复检测3次,并与DNA测序结果进行比较。结果优选的SNaPShot反应的退火温度为55℃;仅需3 h即可对HPV进行分型;在该最适条件下使用R6G-ddATP/SNaPShot-凝胶荧光法检测3种HPV基因型,检测结果与测序结果一致。结论成功建立了3种HR-HPV基因型的微量检测方法——R6G-ddATP/SNaPShot-凝胶荧光法,可用于HPV基因型的快速检测。
文摘Europium ternary complex of Eu(NNA) 3(NNA=α-naphthylacetic acid) was embedded in silica matrix at different ratios by sol-gel method. The luminescence properties of silica composites were studied by comparing them with those of corresponding pure complex by means of excitation, emission spectra and lifetimes. The fluorescence lifetime was prolonged when the pure complex was incorporated in silica matrix. The relative fluorescence intensity and fluorescence lifetimes increased simultaneously with the increase of (Eu(NNA) 3.)
