枯草芽孢杆菌黑色变种芽孢测量方法研究

隋志伟; 王斌; 刘思渊; 王晶; 傅博强; 卓婷烨; 王义

doi:10.3969/j.issn.1000-1158.2020.09.21

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计量学报 ›› 2020, Vol. 41 ›› Issue (9) : 1171-1176. DOI: 10.3969/j.issn.1000-1158.2020.09.21

电离辐射、标准物质与生物计量

枯草芽孢杆菌黑色变种芽孢测量方法研究

隋志伟¹,王斌^1,2,刘思渊¹,王晶¹,傅博强¹,卓婷烨¹,王义²

作者信息 +

Study on Measurement Method of Bacillus subtilis var.niger Spore

Author information +

文章历史 +

摘要

枯草芽孢杆菌黑色变种芽孢是生物防护和灭菌效果评价的重要指示微生物,其量值准确可以保证评价的有效性和可比性。对枯草芽孢杆菌涂布平板法进行了方法验证和协同实验验证以及不确定度分析。结果显示,当平板上的菌落数在30～300CFU之间时,该方法重复性和再现性均较好,室内重复性相对标准偏差为10.0%,室间相对标准偏差为14.1%。经统计分析,胰蛋白胨大豆琼脂培养基平板涂布法相对扩展不确定度为12.0%(k=2),适用于枯草芽孢杆菌黑色变种芽孢定量测量,对于人员防护装备防护性能评价、消毒剂检测及灭菌质量控制具有重要意义。

Abstract

Bacillus subtilis var.niger is an important indicator microbiology of biological protection and sterilization effect evaluation, and its accurate value can ensure the effectiveness and comparability of the evaluation. Spread plate method for bacillus subtilis was method validated, collaborative experimental verified, and uncertainty analyzed. The results showed that spread plate method has a good reproducibility and repeatability, within-laboratory relative standard deviation of repeatability is 10.0%, and inter-laboratory relative standard deviations is 14.1%, when the number of colonies on the plate is between 30CFU and 300CFU. The statistic test showed that the relative expanded uncertainty of spread plate method is 12.0% (k=2). This method can be used for quantitative measurement of bacillus subtilis var. niger spore, which has important significance for the evaluation of protective performance of personnel protective equipment, detection of disinfectants and quality control of sterilization.

导出引用

隋志伟,王斌,刘思渊,王晶,傅博强,卓婷烨,王义. 枯草芽孢杆菌黑色变种芽孢测量方法研究[J]. 计量学报. 2020, 41(9): 1171-1176 https://doi.org/10.3969/j.issn.1000-1158.2020.09.21

Study on Measurement Method of Bacillus subtilis var.niger Spore[J]. Acta Metrologica Sinica. 2020, 41(9): 1171-1176 https://doi.org/10.3969/j.issn.1000-1158.2020.09.21

中图分类号： TB99

参考文献

［1］赵新林, 赵思峰. 枯草芽孢杆菌对植物病害生物防治的作用机理［J］. 湖北农业科学, 2011, 50(15):3025-3028.
Zhao X L, Zhao S F. Research Advance in Controlling Plant Diseases by Bacillus subtilis［J］. Hubei Agricultural Sciences, 2011, 50(15):3025-3028.
［2］张爱武, 薛军. 枯草芽孢杆菌在动物生产中的应用效果［J］. 中国畜牧兽医, 2011, 38(4):234-238.
Zhang A W, Xue J. Effect of Bacillus subtilis in Animal Production［J］. China Animal Husbandry & Veterinary Medicine, 2011, 38(4):234-238.
［3］黄曦, 许兰兰, 黄荣韶, 等. 枯草芽孢杆菌在抑制植物病原菌中的研究进展［J］. 生物技术通报, 2010,(1):24-29.
Huang X, Xu L L, Huang R S, et al. Research Advance in Controlling Plant Diseases by Bacillus subtilis［J］. Biotechnology Bulletin, 2010,(1):24-29.
［4］陈尚智, 胡勇有. 枯草芽孢杆菌对微污染水体的净化作用［J］. 环境科学学报, 2011, 31(8):1594-1601.
Chen S Z, Hu Y Y. Use of Bacillus subtilis in purification of slightly-polluted water［J］. Acta Scientiae Circumstantiae, 2011, 31(8):1594-1601.
［5］刘波, 李红, 姚粟,等. 枯草芽孢杆菌黑色变种ATCC 9372的特性及其应用［J］. 中国消毒学杂志, 2009, 26(2):236-237.
Liu B, Li H, Yao L, et al. Characteristics and application of Bacillus subtilis var. niger ATCC 9372［J］. Chinese Journal of Disinfection, 2009, 26(2):236-237.
［6］NSF/ANSI 49—2016 Biosafety Cabinetry: Design, Construction, Performance, and Field Certification［S］. 2016.
［7］BS EN 12469:2000 Biotechnology-Performance criteria for microbiological safety cabinets［S］. 2000.
［8］YY 0569—2011 Ⅱ级生物安全柜［S］. 2011.
［9］GB/T 26428—2010 饲用微生物制剂中枯草芽孢杆菌的检测［S］. 2010.
［10］GB/T 6379.1—2004 测量方法与结果的准确度(正确度与精密度)第2部分:确定标准测量方法重复性与再现性的基本方法［S］. 2004.
［11］ISO Guide 35:2017 Reference materials—Guidance for characterization and assessment of homogeneity and stability［S］. 2017.
［12］JJF 1343—2012 标准物质定值的通用原则及统计学原理［S］. 2012.
［13］韩永志. 统计学在理化检验中的应用第四讲测量方法的重复性及复现性［J］. 理化检验(化学分册), 1999, 35(12): 571-572.
［14］Parenteral Drug Association.Technical Report No.33:evaluation, validation and implementation of new as microbiological testing methods［R］. PDA, 2000.

［15］全浩, 韩永志. 标准物质及其应用技术［M］. 2版. 北京: 中国标准出版社, 2003.
［16］隋志伟, 刘晓夏, 王晶, 等. 粘质沙雷氏菌测量方法研究［J］. 计量学报, 2018, 39(4): 588-592.
Sui Z W, Liu X X, Wang J, et al. Study on Measurement Method of Serratia marcescens ［J］. Acta Metrologica Sinica, 2018, 39 (4): 588-592.
［17］薛蕾, 林婧, 隋志伟, 等. 大肠杆菌定量检测用标准物质的研制［J］. 计量学报, 2015, 36(6): 652-656.
Xue L, Lin J, Sui Z W, et al. Preparation of Reference Material for Quantitative Detection of Escherichia coli［J］. Acta Metrologica Sinica, 2015, 36(6): 652-656.
［18］陈敏璠, 傅博强, 林婧, 等. 阴沟肠杆菌定量标准物质的研制［J］. 计量学报, 2017, 38 (5): 527-531.
Chen M F, Fu B Q, Lin J, et al. Development of Quantitative Reference Material of Enterobacter Cloacae［J］. Acta Metrologica Sinica, 2017, 38 (5): 527-531.
［19］刘思章, 隋志伟, 张韬, 等. 噬菌体ΦX174标准物质定值方法研究［J］. 计量学报, 2019, 40(3): 511-516.
Liu S Z, Sui Z W, Zhang T, et al. Study on Measurement System for Bacteriophage ΦX174 Reference Material［J］. Acta Metrologica Sinica, 2019, 40(3): 511-516.