Abstract
Shigella is an important human food-borne zoonosis bacterial pathogen, and can cause clinically severe diarrhea. There is an urgent need to develop a specific, sensitive, and rapid methodology for detection of this pathogen. In this study, loop-mediated isothermal amplification (LAMP) combined with magnetic immunocapture assay (IC-LAMP) was first developed for the detection of Shigella in pure culture, artificial milk, and clinical stool samples. This method exhibited a detection limit of 8.7 CFU/mL. Compared with polymerase chain reaction, IC-LAMP is sensitive, specific, and reliable for monitoring Shigella. Additionally, IC-LAMP is more convenient, efficient, and rapid than ordinary LAMP, as it is more efficiently enriches pathogen cells without extraction of genomic DNA. Under isothermal conditions, the amplification curves and the green fluorescence were detected within 30 min in the presence of genomic DNA template. The overall analysis time was approximately 1 h, including the enrichment and lysis of the bacterial cells, a significantly short detection time. Therefore, the IC-LAMP methodology described here is potentially useful for the efficient detection of Shigella in various samples.
Highlights
Shigella is recognized as an important bacterial pathogen worldwide, and may cause several diseases, such as purulent blood, diarrhea, spasms, and shocks, which leads a huge threat to society (Iseki et al, 2007)
loop-mediated isothermal amplification (LAMP) was combined with magnetic IC-LAMP for the detection of Shigella in processed milk and clinical samples (Supplementary Figure S3)
The IC-LAMP methodology was able to detect the pathogen in real-time in a reaction, and the detection and amplification reaction could be conducted simultaneously within 1 h including enrichment of Shigella (Wang et al, 2015)
Summary
Shigella is recognized as an important bacterial pathogen worldwide, and may cause several diseases, such as purulent blood, diarrhea, spasms, and shocks, which leads a huge threat to society (Iseki et al, 2007). Studies have revealed the association between Shigella and most of the bacillary dysentery in developing nations (Iseki et al, 2007; Lin et al, 2010). For use in the food industry and to protect public health, rapid, specific, and sensitive methodologies are required to detect this pathogen (Wang et al, 2015). All kinds of studies have reported that the large invasion plasmid is existed in all Shigella.
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