Ultra-sensitive, high-throughput detection of infectious diarrheal diseases by portable chemiluminescence imaging

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.