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利用载色体chromatophore上的F0F1-ATPase分子马达生物传感器,建立了应用在食品检测中快速检测方法。首先从嗜热菌中提取载色体后,标记荧光探针F-DHPE,合成生物素化单增李氏菌prfA探针,在已标记F-DHPE的载色体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-prfA探针,将待测单核细胞增生李斯特菌标准菌株和阴性对照分别与此生物传感器结合,通过环境H+量测定ATP产生量,进而对单核细胞增生李斯特菌DNA进行检测。结果表明,chroprfA(连接在载色体chro上的prfA探针)的浓度在0.052mg/mL,单核细胞增生李斯特菌DNA浓度在40ng/mL为最适检测条件。通过传统检测方法及PCR检测方法对照,本方法具有良好的检测符合性。
The F0F1-ATPase molecular motor biosensor on chromatophore was used to establish a rapid detection method for food testing. Firstly, the carrier was extracted from thermophilic bacterium, and the fluorescent probe F-DHPE was labeled to synthesize the biotinylated Prolifera listeria prfA probe. The epsilon subunit of the chromosomal ATP synthase of labeled F-DHPE Linked to the epsilon subunit antibody-biotin-streptavidin-biotin-prfA probe, the standard strain of Listeria monocytogenes to be tested and the negative control were respectively bound to the biosensor, and the amount of H + ATP production, and then detect the Listeria monocytogenes DNA. The results showed that the concentration of chroprfA (prfA probe linked to chromate on chromosome) was 0.052 mg / mL, and the concentration of Listeria monocytogenes DNA was 40 ng / mL was the optimal detection condition. Through the traditional detection methods and PCR detection methods control, the method has good detection compliance.