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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,合成生物素化宋内氏志贺氏菌IpaH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-IpaH探针,将待测宋内氏志贺氏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合成10 min后的ATP产生量,进而对宋内氏志贺氏菌DNA进行检测。ATP合成的量可以通过luciferase-luciferin检测试剂所体现的荧光强度大小标定。结果表明,chroIpaH(连接在载色体chro上的IpaH探针)的浓度在0.031 mg/mL,宋内氏志贺氏菌DNA浓度在50 ng/mL为最适检测条件。通过与实际检测样品的传统检测方法及PCR检测方法对照,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. First, the carotenoid was extracted from the thermophilic bacterium to synthesize the biotinylated IpaH probe of Shigella sonnei, and the epsilon subunit of the chromosomal ATP synthase was linked to the epsilon subunit antibody biotin-streptavidin And biotin-IpaH probe. The standard strain of Shigella sonnei to be tested and the negative control were respectively combined with the biosensor to compare the production of ATP after 10 min of ATP synthesis, Shigella DNA was tested. The amount of ATP synthesis can be quantified by the magnitude of the fluorescence reflected by the luciferase-luciferin assay. The results showed that the concentration of chroIpaH (IpaH probe attached to chromatoid chro) was 0.031 mg / mL and the DNA concentration of Shigella sonnei was 50 ng / mL. Through the actual test samples with the traditional detection methods and PCR detection methods control, with good detection of compliance.