Listeria monocytogenes is a gram-positive bacterium that infects individuals after consuming contaminated food and it’s linked to a high mortality rate worldwide. This experiment aimed to determine the gene expression of IL-1β in the small intestine in diarrhea patient exposed to L. monocytogenes. The investigation was carried out using the small intestine of rabbit injected with 1 ml of a 107 CFU/ml of L. monocytogenes, and small intestine of rabbit injected with 1 ml of PBS as a control. Eight hours later of in vivo incubation time, the gene expression level of IL-1β in was determined in the rabbit small intestinal tissue injected by L. monocytogenes using RT-qPCR technique. Result indicated an apparent increase in ILــ1β gene expression level in the rabbit small intestine injected by L. monocytogenes compared to the control. The expression level of ILــ1β was significantly increased with 14.8-fold change in intestinal tissue injected by L. monocytogenes compared to 6.9-fold change in intestinal tissue of the control. This study proposes a strategy for understanding the intestinal tissue immune response to L. monocytogenes invasion.