数字化PCR芯片技术用于食源性细菌多重定量检测OA北大核心CSTPCD
Digital PCR Chip for Multiple Quantitative Detection of Foodborne Bacteria
食源性细菌可造成食物腐败变质并产生多种有害毒素,严重危害食品安全和人体健康.因此,开发食源性细菌有效的检测方法具有十分重要的意义.本研究提出了基于多重数字化聚合酶链式反应(PCR)芯片的分析方法,可对3种常见食源性细菌(金黄色葡萄球菌(Staphylococcus aureus)、沙门氏菌(Salmonella typhimurium)和单增李斯特菌(Listeria monocytogenes))进行快速且高效的检测.此芯片采用离心力驱动的数字化进样方式,15 min内即可完成数字化检测过程,操作简便.PCR实验结果表明,经历25个热循环后即可检出阳性信号.此芯片在低浓度细菌核酸定量检测方面具有显著优势,可有效避免假阴性结果.利用此芯片可对浓度为105～108 CFU/mL的细菌样本进行多重检测.相比荧光定量PCR技术,此芯片在分析低含量靶标时具有更高的准确度和灵敏度.
The presence of food-borne pathogenic bacteria poses a severe threat to human health. Therefore,it is essential to develop effective methods for bacteria detection. Herein,a method based on digital polymerase chain reaction (PCR) chip was developed for rapid and effective detection of three kinds of common food-borne bacteria (Staphylococcus aureus,Salmonella typhimurium and Listeria monocytogenes). Driven by centrifugal force,samples could be easily loaded and digitized in 15 min. PCR results showed that the positive microchambers in the chip could be identified after 25 thermal cycles. The chip had advantages in the quantitative detection of bacterial nucleic acid at low concentration,which could effectively avoid false negative results. Multiple detection of bacterial samples with concentration of 105-108 CFU/mL could also be realized using this chip. Compared with fluorescent quantitative PCR,this method had higher accuracy and sensitivity,especially for detection of targets at low concentration.
茅珍珍;胡文琪;唐曲;蒋汶君;秦玉岭;吴丽
南通大学公共卫生学院,南通市公共卫生与医学分析重点实验室,南通226019南通大学医学院,南通226001
数字化芯片多重聚合酶链式反应金黄色葡萄球菌沙门氏菌单增李斯特菌
Digital chipMultiple polymerase chain reactionStaphylococcus aureusSalmonellaListeria monocytogenes
《分析化学》 2024 (006)
789-798 / 10
国家自然科学基金项目(No.31901056)和江苏省杰出青年基金项目(No.BK20220060)资助.Supported by the National Natural Science Foundation of China(No.31901056)and the Excellent Youth Foundation of Jiangsu Scientific Committee(No.BK20220060).
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