目的: 改良细菌生长曲线测定方法,系统评估检测设备(分光光度计vs酶标仪)、培养方式(集中vs试管)及检测时序(实时vs终点)对细菌生长曲线测定的影响。方法: 以大肠埃希菌和金黄色葡萄球菌为模式菌株,采用三因素正交实验设计(8种组合),通过分光光度计(OD600)与酶标仪(OD630)测定光密度值,绘制生长曲线并分析设备间相关性、时序等效性及培养稳定性。结果: 不同方法组合的生长曲线呈高度正相关(P＜0.01);实时检测与冷藏终点检测的OD值比较差异无统计学意义(P＞0.05)。优化后的试管培养-终点酶标仪(OD630)检测法通过预混接种标准化和单管连续取样,使曲线平滑度明显提升。结论: 基于试管预混培养、4 ℃冷藏终点检测及酶标仪高通量分析(OD630)的组合方法,可同步提升操作效率与数据一致性,为细菌生长曲线测定提供优化方案。
Objective: To optimize the bacterial growth curve determination method and systematically evaluate the effects of detection devices (spectrophotometer vs. microplate reader), culture methods (batch vs. aliquoted tube), and detection timing (real-time vs. endpoint) on growth curve analysis. Methods: Using Escherichia coli and Staphylococcus aureus as model strains, a three-factor orthogonal experimental design (8 combinations) was implemented. Optical density values were measured via spectrophotometer (OD600) and microplate reader (OD630) to plot growth curves. The growth curve was drawn and the correlation between devices, time sequence equivalence and culture stability were analyzed. Results: Growth curves under different method combinations showed strong positive correlations (P＜0.01). No significant differences were observed between real-time and endpoint refrigeration detection in OD values (P＞0.05). The optimized tube culture-endpoint-microplate reader (OD630) detection method significantly improved the smoothness of the curve by pre-mixed inoculation standardization and single-tube continuous sampling. Conclusion: The integrated protocol combining tube pre-mixed culture, 4 ℃ refrigeration endpoint detection, and microplate reader-based high-throughput analysis (OD630) improves operational efficiency and data consistency, providing an optimization scheme for bacterial growth curve determination.
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