Objective To develop an enzyme-linked immunochromatographic test strip for the determination of amanita toxin in mushroom based on immunochromatography. Methods N-hydroxysuccinimide (NHS) and N-(3-dimethylaminopropyl)-N’-ethylcarbodiimide hydrochloride (EDC) were used as activators, 6-aminohexanoic acid was introduced to obtain amanitin peptide hapten, and further conjugated with keyhole limpet hemocyanin (KLH) and bovine serum albumin (BSA) to prepare immunogen and coating antigen. Then, the immunogen was used to immunize Balb/c mice to prepare monoclonal antibodies. The colloidal gold immunochromatographic test strip for amanitin was successfully developed by preparing colloidal gold and gold labeled antibody, optimizing the concentration of coating antigen, secondary antibody and extracting solution, and then its cross reactivity, stability, repeatability, and accuracy were evaluated. Results The limits of detection of α-amatoxin, β-amatoxin, and γ-amatoxin peptide were 100, 20 and 50 μg/kg, respectively. The sensitivity was 100%, the false positive rate and false negative rate were 0%, and the results were consistent with the existing instrumental methods without cross reaction with other mushroom toxins. Conclusion The test strip has the characteristics of simple operation, high repeatability and good stability, and is suitable for on-sites creening and rapid detection of amanitin in mushrooms.
| 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 |