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Salmonella  is a pathogenic Gram-negative bacterium and important zoonotic pathogen survive in host cell, which is well known to cause gastrointestinal infections and even death in human through food contamination.  Salmonella  survives and reproduces within cells in two forms of cytosolic  Salmonella  and vacuolar  Salmonella . However, the current researches on the mechanism of  Salmonella  infection was mostly based on the total numbers, and there is no clear distinction between cytosolic  Salmonella  and vacuolar  Salmonella . This article combined biotin-streptavidin-amplified enzyme-linked immunosorbent assay(BA-ELISA) and Flat colony counting method to separate and quantify cytosolic  Salmonella  and vacuolar  Salmonella.  The results showed that the anti- Salmonella  antibody with a dilution ratio of 1:500 can bind ~1700  Salmonella , and the binding rate reached to~70%. In terms of bacterial counts, vacuolar  Salmonella  dominates in the early stage of  Salmonella  infection, and cytosolic  Salmonella  dominates in the later stage, the reproduction rate of vacuolar  Salmonella  is significantly lower than cytosolic  Salmonella  during  Salmonella  infection, the dynamic growth curve of cytosolic  Salmonella  and vacuolar  Salmonella  intuitively reflected the proliferation status of the two forms of  Salmonella  during the entire process of  Salmonella  infection.

BA-ELISA isolation of cytosolic Salmonella and vacuolar Salmonella