梯度金黄色葡萄球菌DNA杂交分子信标激发荧光初探

Exploration for Different Concentrations of Staphylococcus aureus DNA Hybridize with Molecular Beacon that Motivates Fluorescence

DOI：10.3969/j.issn.1673-1689.2014.01.011

中文关键词: 分子信标 梯度DNA 金黄色葡萄球菌

英文关键词: beacon different concentration DNA Staphylocossus aureus

基金项目:国家质量监督检验检疫总局公益性行业科研专项项目(201310130);长沙市科技计划项目(K1203030-71)

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中文摘要:

验证不同浓度黄色葡萄球菌DNA与特殊设计的分子信标探针杂交,观察荧光强度的变化,并验证该探针对于金黄色葡萄球菌DNA的特异性。利用特殊ATMND染料特异性嵌入缺失DNA位点的独特分子信标设计,并巧妙利用了能与DNA茎环结构杂交互补金黄色葡萄球菌特异DNA序列,导致茎环结构构相改变,而使ATMND染料离开缺失位点被重新激发荧光。将不同浓度黄色葡萄球菌DNA与分子信标探针杂交,观察荧光强度的变化,并选择标准菌株DNA对该探针进行特异性验证。证实了这种探针能够特异性的选择杂交金黄色葡萄球菌目标序列,随着金黄色葡萄球菌DNA浓度的增强,荧光信号也随之增强,与其它沙门氏菌、大肠杆菌、阪崎肠杆菌、单核细胞增生李斯特菌和铜绿假单胞菌DNA相比,具有更为明显的荧光恢复功能。荧光信号随着金黄色葡萄球菌DNA浓度的增高而增强,该探针在金黄色葡萄球菌DNA的识别上具有一定的特异性。

英文摘要:

In this manuscript,the effect of Staphylococcus aureus DNA hybridize special molecular beacon on the fluorescence intensity was determined to designed a probe to sensitive to Staphylococcus aureus DNA. The method using unique molecular beacon combined with ATMND dye which embed into absent site to hybridize complementary Staphylococcus aureus specific DNA sequence,resulting in stem-loop structure phase change and the ATMND dye is excited fluorescence. Different concentration of Staphylococcus aureus DNA hybrid with designed molecular beacon and observe the changes of fluorescence intensity. Finally,the standard strains DNA were used to verify the sensitivity to the probe. The probe has high sensitive to Staphylococcus aureus target DNA,along with high concentration of target DNA. Compared with other Salmonella,Escherichia coli,Enterobacter sakazakii,Listeria monocytogenes and Pseudomonas aeruginosa,the probe has more obvious fluorescence recovery ability. The fluorescence signal increased along with the increase of Staphylococcus aureus DNA concentration,the probe has a certain specificity in identification of Staphylococcus aureus DNA.

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