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This study aimed to establish a rapid and effective indirect ELISA method for detecting Listeria monocytogenes infection in samples. The InlB protein was expressed and purified in vitro using a prokaryotic expression system. The optimal parameters for the indirect ELISA method were determined through optimization， an indirect ELISA method was established. Soluble expression of InlB protein was achieved using the pCold II-10 × His vector. Optimal conditions determined by a grid-screen method were： InlB protein coating concentration of 4 μg·mL-1， blocking with 10% skim milk powder at room temperature for 2 hours， optimal serum dilution of 1∶400， incubation at 37 ℃ for 1 hour， the optimal dilution for the secondary antibody was 1∶2 500， with incubation at 37 ℃ in the dark for 8 min. The positive/negative cutoff value was determined to be 0.37. The indirect ELISA method established in this study reacted only with L. monocytogenes-positive sera， maintaining positive detection at a serum dilution of 1∶800. The coefficient of variation （CV） for intra-batch replicates ranged from 2.79% to 6.64%， while the CV for inter-batch replicates ranged from 2.61% to 7.75%. Compared with a commercial kit， the overall agreement rate was 95.83%. The indirect ELISA method established in this study for detecting L. monocytogenes-infected samples exhibits high specificity， sensitivity， and reproducibility. It can be used for rapid detection of L.monocytogenes-infected samples， providing a reference for monitoring listeriosis.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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