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本研究建立了应用F0F1-ATPase分子马达生物传感器快速检测阪崎肠杆菌的方法。自嗜热菌中提取载色体后,合成针对阪崎肠杆菌的生物素化ITS探针,在载色体ATP合酶的ε亚基上连接ε亚基抗体-生物素-链霉亲和素-生物素-ITS探针,将待测阪崎肠杆菌标准菌株和阴性对照分别与此生物传感器结合,比较其催化ATP产生量,进而对阪崎肠杆菌DNA进行检测。结果表明,chro ITS探针浓度0.019mg/mL,阪崎肠杆菌DNA浓度40ng/mL为最适检测条件。通过与传统检测方法及PCR检测方法对照,本方法具有良好的检测符合性。
This study established a F0F1-ATPase molecular motor biosensor for rapid detection of Enterobacter sakazakii method. After extracting the chromosome from the thermophilic bacterium, the biotinylated ITS probe for Enterobacter sakazakii was synthesized, and the epsilon subunit antibody-biotin-streptavidin was linked to the epsilon subunit of the chromosomal ATP synthase Biotin-ITS probe, Enterobacter sakazakii standard strains to be tested and the negative control were combined with the biosensor, compared with its catalytic ATP production, and then Enterobacter sakazakii DNA was detected. The results showed that chro ITS probe concentration of 0.019mg / mL, Enterobacter sakazakii DNA concentration of 40ng / mL is the best detection conditions. Compared with the traditional detection methods and PCR detection methods, this method has good detection of compliance.