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
By the electrochemical anodization method,we achieve the single-layer macroporous silicon on the N-type silicon,and prepare gold nanoparticles with sodium citrate reduction method. Through injecting the gold nanoparti...By the electrochemical anodization method,we achieve the single-layer macroporous silicon on the N-type silicon,and prepare gold nanoparticles with sodium citrate reduction method. Through injecting the gold nanoparticles into the porous silicon by immersion,the fluorescence quenching mechanism of porous silicon influenced by gold nanoparticles is analyzed. Then the macroporous silicon deposited with gold nanoparticles is utilized to enhance the fluorescence of rhodamine 6G(R6G). It is found that when the macroporous silicon is deposited with gold nanoparticles for 6 h,the maximum fluorescence enhancement of R6G(about ten times) can be realized. The N-type porous silicon deposited with gold nanoparticles can be an excellent substrate for fluorescence detection.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。
基金supported by the National Natural Science Foundation of China(Nos.61308120,61575168 and 11264038)the Doctor Startup Project of Xinjiang University(No.BS120122)+1 种基金the Young Talents Project in Xinjiang Uygur Autonomous Region(No.2013731003)the Xinjiang Science and Technology Project(Nos.2015211C262 and 2014211B003)
文摘By the electrochemical anodization method,we achieve the single-layer macroporous silicon on the N-type silicon,and prepare gold nanoparticles with sodium citrate reduction method. Through injecting the gold nanoparticles into the porous silicon by immersion,the fluorescence quenching mechanism of porous silicon influenced by gold nanoparticles is analyzed. Then the macroporous silicon deposited with gold nanoparticles is utilized to enhance the fluorescence of rhodamine 6G(R6G). It is found that when the macroporous silicon is deposited with gold nanoparticles for 6 h,the maximum fluorescence enhancement of R6G(about ten times) can be realized. The N-type porous silicon deposited with gold nanoparticles can be an excellent substrate for fluorescence detection.