目的 采用“细菌计数－内毒素定量－电镜形态”多维评价体系，比较过氧乙酸、次氯酸钠、酶清洗剂、碱洗及蒸汽5种方法对透析供水管路生物膜的清除效能。 方法 在316 L不锈钢载体上建立铜绿假单胞菌(pseudomonas aeruginosa ATCC 27853)成熟生物膜模型。将样本分对照组、0.2%过氧乙酸、0.2%次氯酸钠、酶清洗剂、碱洗、热蒸汽6组处理，每组独立重复3次。处理后，采用Luria-Bertani(LB)琼脂培养基与寡营养的Reasoner's 2A(R2A)琼脂培养基检测细菌残留，计算对数减少率(log reduction value，LRV)；采用动态浊度法测定内毒素LRV；并通过扫描电镜(scanning electron microscopy，SEM)验证物理清除效果。 结果 ①细菌去除效果：对照组LB培养基菌数为(1.37±1.10)×107 CFU/ml。各组间LB-LRV总体比较差异有统计学意义(F=64.330，P＜0.001)。热蒸汽组在2种培养基中均未检出细菌(LB-LRV＞7.14)；过氧乙酸组与次氯酸钠组的杀菌效果与热蒸汽组相比，差异均无统计学意义(t=1.017、2.580，P=0.842、0.148)。②内毒素去除效果：对照组内毒素量为(1.14±1.05)×104 EU/ml，各处理组内毒素LRV总体比较差异有统计学意义(F=10.480，P=0.001)。热蒸汽组清除效能(LRV=2.23)优于过氧乙酸组(t=4.571，P=0.007)、酶清洗剂组(t=4.636，P=0.006)及碱洗组(t=5.171，P=0.003)。③电镜形态：过氧乙酸组、次氯酸钠组和热蒸汽组可见生物膜碎片；过氧乙酸组可见细菌皱缩但基质残留；酶清洗剂组与碱洗组仍见大量完整细菌或细胞聚集。 结论 去除生物膜5组方法中，蒸汽组在杀菌、去内毒素与物理剥离上都优于其他组，次氯酸钠在有效杀灭细菌的同时，呈现出较好的内毒素清除与基质剥离趋势，达到次优。单一酶清洗剂和碱洗消毒对生物膜去除效果差。虽然部分消毒方法在细菌检测结果方面达到透析用水质控标准，但5种单一消毒方法去除生物膜内毒素均未达到透析用水标准。

Objective To compare the efficacy of five disinfection methods, including peracetic acid, sodium hypochlorite, enzymatic detergent, alkaline wash, and steam, for biofilm removal in hemodialysis water supply pipelines using a multidimensional evaluation system involving "bacterial count - endotoxin quantification - electron microscopic morphology".  Methods  A mature biofilm model of Pseudomonas aeruginosa (ATCC 27853) was established on 316L stainless steel carriers. The samples were divided into six treatment groups: a control group, 0.2% peracetic acid, 0.2% sodium hypochlorite, enzymatic detergent, alkaline wash and hot steam, with each group repeated independently three times. After treatment, bacterial residues were detected using both Luria-Bertani (LB) agar and Reasoner's 2A (R2A) low-nutrient agar to calculate the Log Reduction Value (LRV); endotoxin LRV was determined by kinetic turbidimetric assay; and physical removal was verified by scanning electron microscopy (SEM).  Results  ①Bacterial removal: The bacterial count in the control group on LB media was (1.37±1.10)×107 CFU/ml. There was a statistically significant difference in the overall LB-LRV values between the groups (F=64.330, P＜0.001). No bacteria were detected in the hot steam group in either of the two culture media (LB-LRV >7.14); there were no statistically significant differences in the bactericidal efficacy of the peracetic acid group and the sodium hypochlorite group compared with the hot steam group (t=1.017, 2.580; P=0.842, 0.148).  ②Endotoxin removal efficacy: The endotoxin concentration in the control group was (1.14±1.05)×104 EU/ml. Overall differences in endotoxin LRV among treatment groups were statistically significant (F=10.480, P=0.001). The hot steam group demonstrated superior removal efficacy (LRV=2.23) compared with the peracetic acid group (t=4.571, P=0.007), the enzymatic detergent group (t=4.636, P=0.006) and the alkaline detergent group (t=5.171, P=0.003).  ③Electron microscopic morphology: Fragments of biofilm were observed in the peracetic acid group, the sodium hypochlorite group and the hot steam group; in the peracetic acid group, bacteria appeared shrivelled but matrix residues remained; in the enzymatic detergent group and the alkaline detergent group, a large number of intact bacteria or cell aggregates were still observed.  Conclusions  Among the five biofilm removal methods, the steam group was significantly superior to other groups in sterilization, endotoxin removal, and physical detachment. Sodium hypochlorite achieved effective bacterial eradication while exhibiting favorable trends in endotoxin clearance and substrate detachment, yielding the second-best results. Although some disinfection methods met the bacterial quality control standards for dialysis water, none of the five single disinfection methods met the dialysis water standards for biofilm endotoxin removal.