title: Ultra-sensitive, high-throughput detection of infectious diarrheal diseases by portable chemiluminescence imaging.
abstract: This paper describes a rapid, ultra-sensitive, and high-throughput pathogenic DNA identification strategy for infectious diarrheal diseases diagnosis. This strategy is based on specific DNA hybridization and horseradish-peroxidase-catalyzed chemiluminescence (CL) detection. Probe DNA strands are covalently immobilized on the aldehyde-group-modified slide and hybridized with biotin-modified target DNA strands. Horseradish-peroxidase (HRP) is then combined with the target DNA via a biotin-streptavidin linkage. The subsequently added mixture of luminol and hydrogen peroxide is catalyzed by HRP and radiates photons. The photons are collected and read out by a portable imager. The specific detection of target DNA strands was realized at a detection limitation of about 0.75 nM. This strategy facilitates quantitative detection, as indicated by the fact that the CL signals were consistent well with a linear function. This method was applied to identify a myriad of real diarrheal pathogens samples, including Enterohemorrhagic Escherichia coli (EHEC), Vibrio cholerae (VBC), Shigella (SHLA), and Salmonella (SMLA). Triple-assay of six gene sequences from these pathogens was realized, which facilitates accurate, high-throughput identification of diarrheal pathogens. This CL assay strategy is appropriate for application in disease diagnosis and prevention.
authors.affiliation: Key Laboratory of New Molecular Diagnosis Technologies for Infectious Diseases, Institute of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850, PR China; College of Mechatronics and Automation, National University of Defense Technology, Changsha, Hunan 410073, PR China.
authors.forename: Chaoguang
authors.initials: C
authors.lastname: Wang
authors.affiliation: Key Laboratory of New Molecular Diagnosis Technologies for Infectious Diseases, Institute of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850, PR China.
authors.forename: Rui
authors.initials: R
authors.lastname: Xiao
authors.affiliation: College of Mechatronics and Automation, National University of Defense Technology, Changsha, Hunan 410073, PR China; State Key Laboratory of Transducer Technology, Chinese Academy of Science, Shanghai 200050, PR China. Electronic address: ptdong@nudt.edu.cn.
authors.forename: Peitao
authors.initials: P
authors.lastname: Dong
authors.affiliation: College of Mechatronics and Automation, National University of Defense Technology, Changsha, Hunan 410073, PR China.
authors.forename: Xuezhong
authors.initials: X
authors.lastname: Wu
authors.affiliation: Key Laboratory of New Molecular Diagnosis Technologies for Infectious Diseases, Institute of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850, PR China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Rong
authors.affiliation: Key Laboratory of New Molecular Diagnosis Technologies for Infectious Diseases, Institute of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850, PR China.
authors.forename: Lin
authors.initials: L
authors.lastname: Xin
authors.affiliation: Benxi County Hospital of Traditional Chinese Medicine, Benxi 117100, PR China.
authors.forename: Jun
authors.initials: J
authors.lastname: Tang
authors.affiliation: Key Laboratory of New Molecular Diagnosis Technologies for Infectious Diseases, Institute of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850, PR China. Electronic address: sqwang@bmi.ac.cn.
authors.forename: Shengqi
authors.initials: S
authors.lastname: Wang
mesh_terms: D015374:Biosensing Techniques / Q000191:economics / Q000379:methods*; D004269:DNA, Bacterial / Q000302:isolation & purification*; D004403:Dysentery / Q000175:diagnosis* / Q000382:microbiology*; D057166:High-Throughput Screening Assays / Q000191:economics / Q000379:methods; D006801:Humans; D057230:Limit of Detection; D008163:Luminescent Measurements / Q000191:economics / Q000379:methods*; D009693:Nucleic Acid Hybridization / Q000379:methods; D019095:Point-of-Care Systems / Q000191:economics
chemical_list: D004269:DNA, Bacterial
keywords: Chemiluminescence; DNA; High-throughput; Imager; Infectious diarrheal disease