不锈钢载体型过氧化氢蒸汽灭菌生物指示剂芽孢计数方法的建立  

Establishment of spore counting methods of stainless steel biological indicator for vaporized hydrogen peroxide sterilization

作　　者：邓景韬[1,2] 赵丹萍 代莎 严东珍 刘菊[1] 罗辉艳[1] Deng Jingtao;Zhao Danping;Dai Sha;Yan Dongzhen;Liu Ju;Luo Huiyan(Quality Control Department of Chengdu Institute of Biological Products Co.,Ltd.,Chengdu 610023,China;Sichuan Vaccine Engineering Technology Research Center,Chengdu 610023,China)

机构地区：[1]成都生物制品研究所有限责任公司质量检定室,成都610023 [2]四川省疫苗工程技术研究中心,成都610023

出　　处：《国际生物制品学杂志》2022年第6期334-337,共4页International Journal of Biologicals

摘　　要：目的建立不锈钢载体型过氧化氢蒸汽灭菌生物指示剂总芽孢计数的方法。方法采用不同洗脱液及不同超声参数组合充分洗脱不锈钢载体上的芽孢,并对其进行激活、培养和计数,以考察生物指示剂的芽孢回收率;通过重复性实验确定其最优试验参数,建立总芽孢计数检测方法,并对该方法进行精密度和重现性验证。结果以含0.1%吐温80的灭菌纯化水作为洗脱液,超声频率40 kHz、功率500 W为超声条件作用50 min,芽孢回收率最高可达135.6%,确定该实验参数组合为样品最佳处理条件;3名实验人员分别对3批生物指示剂开展重复性实验,其精密度相对标准偏差在4.0%~14.8%范围内,重现性相对标准偏差分别为8.4%、11.6%和13.8%,该方法的精密度和重现性达到中国药典2020年版的要求。结论研究建立的检测方法可用于不锈钢载体型过氧化氢蒸汽灭菌生物指示剂芽孢载量计数。Objective To establish the test method of total spore count of stainless steel biological indicator for vaporized hydrogen peroxide sterilization.Methods The spores on the stainless steel indicator were eluted sufficiently under different elution conditions and different ultrasonic parameter combinations.The spores were activated,cultured and counted so as to investigate the recovery rate of biological indicator spores.The optimal experiment parameters for the total spore count test method were established through repeated trials and validated with precision and reproducibility.Results The optimal spore recovery rate of 135.6%was obtained using ultrasonic frequency of 40 kHz and power of 500 W,and sterilized purified water containing 0.1%Tween 80 as eluent for 50 min.This combination of experimental parameters was determined to be the best sample processing condition.The repeated study was conducted by 3 operators,each of whom tested 3 batches of the indicator,respectively.The relative standard deviations(RSDs)of the precision were 4.0%-14.8%,and the RSDs of the reproducibility were 8.4%,11.6%and 13.8%,respectively.The precision and reproducibility of this method met the requirements of Chinese pharmacopeia 2020 edition.Conclusion The method established can be used for total spore count of stainless steel biological indicator for vaporized hydrogen peroxide sterilization.

关 键 词：生物指示剂 芽孢回收率 超声参数 洗脱液 

分 类 号：Q93-332[生物学—微生物学]