Gentry M K, Dalrymple J M
J Clin Microbiol. 1980 Sep;12(3):361-6. doi: 10.1128/jcm.12.3.361-366.1980.
The cytotoxic activity of Shigella dysenteriae 1 was assayed by exposing HeLa cells in microtiter cultures to dilutions of toxin. Exposure to toxin caused either failure of cells in suspension to attach or detachment of cells from established monolayers. Estimates of toxin potency were made by staining residual cells with crystal violet and visually inspecting the stained plates. Quantitation of the cytotoxic effect was made possible by eluting and spectrophotometrically measuring the stain. The dilution of toxin causing 50% cell detachment, the endpoint chosen for the assay, was estimated from plots of dye absorbance versus toxin dilution. The 50% cell detachment dilution of toxin varied as a function of cell concentration, incubation of toxin with cells in suspension or as established monolayers, and the cell line used for assay. The HeLa cell line was the most sensitive of the cell lines examined. The method was easily utilized to monitor toxin purification and to measure antitoxin neutralization of toxin activity.
通过在微量培养板中将HeLa细胞暴露于不同稀释度的毒素来检测痢疾志贺氏菌1的细胞毒性活性。暴露于毒素会导致悬浮细胞无法附着或已形成单层的细胞脱离。通过用结晶紫对残留细胞进行染色并目视检查染色后的培养板来估计毒素效价。通过洗脱并分光光度法测量染色剂，实现了对细胞毒性作用的定量。从染料吸光度与毒素稀释度的曲线图中估计导致50%细胞脱离的毒素稀释度，该终点被选作检测指标。毒素导致50%细胞脱离的稀释度随细胞浓度、毒素与悬浮细胞或已形成单层的细胞的孵育情况以及用于检测的细胞系而变化。在所检测的细胞系中，HeLa细胞系最为敏感。该方法易于用于监测毒素纯化以及测量抗毒素对毒素活性的中和作用。
