Yong Li: Sino-UAE International Cooperative Joint Laboratory of Pathogenic Microorganism Rapid Detection, Qingdao Key Laboratory of Nucleic Acid Rapid Detection, Qingdao Nucleic Acid Rapid Detection Engineering Research Center, College of Biological Engineering, Qingdao University of Science and Technology, 266042 Qingdao, China. ORCID
Yanwen Qi: Sino-UAE International Cooperative Joint Laboratory of Pathogenic Microorganism Rapid Detection, Qingdao Key Laboratory of Nucleic Acid Rapid Detection, Qingdao Nucleic Acid Rapid Detection Engineering Research Center, College of Biological Engineering, Qingdao University of Science and Technology, 266042 Qingdao, China.
Jiaqi Liu: Sino-UAE International Cooperative Joint Laboratory of Pathogenic Microorganism Rapid Detection, Qingdao Key Laboratory of Nucleic Acid Rapid Detection, Qingdao Nucleic Acid Rapid Detection Engineering Research Center, College of Biological Engineering, Qingdao University of Science and Technology, 266042 Qingdao, China.
Pengyu Wang: Sino-UAE International Cooperative Joint Laboratory of Pathogenic Microorganism Rapid Detection, Qingdao Key Laboratory of Nucleic Acid Rapid Detection, Qingdao Nucleic Acid Rapid Detection Engineering Research Center, College of Biological Engineering, Qingdao University of Science and Technology, 266042 Qingdao, China.
Jiayu Zheng: Sino-UAE International Cooperative Joint Laboratory of Pathogenic Microorganism Rapid Detection, Qingdao Key Laboratory of Nucleic Acid Rapid Detection, Qingdao Nucleic Acid Rapid Detection Engineering Research Center, College of Biological Engineering, Qingdao University of Science and Technology, 266042 Qingdao, China.
Xiangyu Chen: Sino-UAE International Cooperative Joint Laboratory of Pathogenic Microorganism Rapid Detection, Qingdao Key Laboratory of Nucleic Acid Rapid Detection, Qingdao Nucleic Acid Rapid Detection Engineering Research Center, College of Biological Engineering, Qingdao University of Science and Technology, 266042 Qingdao, China.
Ye Wang: University of Health and Rehabilitation Sciences, Qingdao Central Hospital, 266042 Qingdao, China.
Xiaowen Zhao: University of Health and Rehabilitation Sciences, Qingdao Central Hospital, 266042 Qingdao, China.
Yingqiu Xie: Department of Biology, School of Sciences and Humanities, Nazarbayev University, Astana 010000, Kazakhstan.
Chao Shi: Qingdao Nucleic Acid Rapid Testing International Science and Technology Cooperation Base, College of Life Sciences, Department of Pathogenic Biology, School of Basic Medicine, Department of the Clinical Laboratory, The Affiliated Hospital of Qingdao University, Qingdao University, 266071, Qingdao, China.
Cuiping Ma: Sino-UAE International Cooperative Joint Laboratory of Pathogenic Microorganism Rapid Detection, Qingdao Key Laboratory of Nucleic Acid Rapid Detection, Qingdao Nucleic Acid Rapid Detection Engineering Research Center, College of Biological Engineering, Qingdao University of Science and Technology, 266042 Qingdao, China. ORCID
Rapid and precise identification of the pathogens causing sepsis remains a significant diagnostic challenge. Blood culture is time-consuming and insensitive, while molecular diagnostic techniques, such as the polymerase chain reaction (PCR), are fast but greatly influenced by template quality. Here, we present a new approach to separate trace amounts of pathogen DNA from blood, which utilizes lysozyme to destroy bacteria and release DNA, followed by enrichment and purification using magnetic nanoparticles (MNPs) modified with kanamycin (Kan) or tobramycin (TM). We demonstrate that the prepared Kan@MNPs and TM@MNPs can efficiently adsorb DNA, with the mechanism involving interaction with the minor groove of DNA. Notably, the adoption of lysozyme ensures bacterial lysis while avoiding damage to blood cells, minimizing the interference from human genomic DNA background and inhibitory components, thereby obtaining relatively pure bacterial DNA. For artificially infected whole blood samples, our method shortens the sample processing time to 35 min and achieves a 10-fold improvement in PCR sensitivity compared to a commercial kit. Through clinical evaluation of blood samples collected from suspected infected patients, we identified positive samples that were 100% consistent with the clinical practice. Therefore, this method holds promising potential for clinical application in advancing rapid sepsis diagnosis and earlier interventions.
