化学
单核细胞增生李斯特菌
重组酶聚合酶扩增
核酸
清脆的
聚合酶链反应
核酸扩增试验
计算生物学
细菌
生物化学
病毒学
基因
遗传学
生物
沙眼衣原体
作者
Yiran Xiao,Hong-Lin Ren,Han Wang,De-Ying Zou,Yixin Liu,Haosong Li,Pan Hu,Yansong Li,Zeng-Shan Liu,Shiying Lu
出处
期刊:Talanta
[Elsevier]
日期:2023-07-01
卷期号:259: 124558-124558
被引量:4
标识
DOI:10.1016/j.talanta.2023.124558
摘要
Listeria monocytogenes (LM) is an important foodborne pathogen that is associated with a high mortality rate. Currently, there is an urgent need for an inexpensive and rapid assay for the large-scale diagnosis and monitoring of LM. To meet these requirements, we designed a one-step, low-cost platform for the simultaneous amplification and detection of LM based on the CRISPR/Cas12a system with a micro-amplification (named Cas12a-MA). This method utilizes a combination of CRISPR/Cas12a and recombinase polymerase amplification (RPA) in the same vessel to provide a contamination-free platform for rapid nucleic acid detection with high specificity and ultra-sensitivity. In this study, we screened for three specific genes and selected the hly gene in LM as the final target. Our data showed that the number of amplification products plays a crucial role in the function of the CRISPR/Cas12a system. Our method was then further optimized for the specific detection of target DNA on 4.4 CFU/g in 25min. These assays successfully detected LM in spiked pork samples and natural meat samples (pork, beef, and mutton). All results indicate that Cas12a-MA shows great promise for foodborne pathogen detection.
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