Gu Aoting, Dong Yongzhen, Li Letian, Yu Deyang, Zhang Jiangjiang, Chen Yiping
College of Food Science and Technology, Huazhong Agricultural University, Shizishan Street, Hongshan District, Wuhan, Hubei 430070, China.
State Key Laboratory of Marine Food Processing and Safety Control, Dalian Polytechnic University, Dalian, Liaoning 116034, China.
J Agric Food Chem. 2024 Sep 11;72(36):20130-20139. doi: 10.1021/acs.jafc.4c05540. Epub 2024 Aug 28.
We combined a CRISPR/Cas12a system with a hybridization chain reaction (HCR) to develop an ultrasensitive magnetic relaxation switching (MRS) biosensor for detecting viable (). Magnetic nanoparticles of two sizes (30 and 1000 nm: MNP and MNP, respectively) were coupled through HCR. The gene-activated CRISPR/Cas12a system released MNP from the MNP-HCR-MNP complex through a trans-cleavage reaction. After magnetic separation, released MNP was collected from the supernatant and served as a transverse relaxation time (T) signal probe. Quantitative detection of is achieved by establishing a linear relationship between the change in T and the target gene. The biosensor's limit of detection was 77 CFU/mL (LOD = 3/, = 22.30, = 0.87), and the linear range was 10-10 CFU/mL. The accuracy for detecting in real samples is comparable to that of qPCR. Thus, this is a promising method for the rapid and effective detection of foodborne pathogens.
我们将CRISPR/Cas12a系统与杂交链式反应（HCR）相结合，开发出一种用于检测活（）的超灵敏磁弛豫开关（MRS）生物传感器。两种尺寸的磁性纳米颗粒（分别为30和1000 nm：MNP和MNP）通过HCR耦合。基因激活的CRISPR/Cas12a系统通过反式切割反应从MNP-HCR-MNP复合物中释放出MNP。磁分离后，从上清液中收集释放的MNP，并将其用作横向弛豫时间（T）信号探针。通过建立T变化与靶基因之间的线性关系来实现对的定量检测。该生物传感器的检测限为77 CFU/mL（LOD = 三分之三， = 22.30， = 0.87），线性范围为10-10 CFU/mL。在实际样品中检测的准确性与qPCR相当。因此，这是一种用于快速有效检测食源性病原体的有前景的方法。 （注：原文中存在部分括号缺失内容，翻译时保留了原文格式）
