鼠疫菌噬菌体效价噬菌斑测定法的建立
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摘要
目的建立鼠疫菌噬菌体噬菌斑效价测定方法。方法通过分析细菌接种浓度、孵育吸附时间及培养温度等参数,建立鼠疫菌噬菌体效价测定方法,并分析其精密性;建立鼠疫活疫苗鉴别及纯菌检查用噬菌体效价质量标准。结果经优化后确定细菌接种浓度为7×108/mL,不需孵育吸附,培养温度为29℃,所建立的检测方法精密性较好,用于鼠疫活疫苗鉴别及纯菌检查用噬菌体效价质量标准应不低于1×106PFU/mL。结论建立了鼠疫菌噬菌体噬菌斑效价测定方法,为鼠疫菌噬菌体及疫苗质量控制奠定了基础。
Objective To develop a plaque assay in determination of titer of phage for Yersinia pestis. Methods It is by optimizing the concentration of Yersinia pestis cells for inoculation,adsorption time and culture temperature,the plaque assay was developed in determination of titer of phage for Yersinia pestis and estimated precision for the developed method. Quality standard of titer was established through identity test and bacterial purity test. Results The optimal cell concentration of Yersinia pestis for inoculation is 7 × 108cells / mL,adsorption is not necessary,and culture temperature is 29 ℃. It shows a good precision by the developed method. Quality standard for titer of phage is no less than 106PFU / mL. Conclusion A plaque assay is developed in determination titer of phage for plaque of Yersinia pestis used in detection of plague vaccine and quality control,and quality standard for titer of phage is established too.
Objective To develop a plaque assay in determination of titer of phage for Yersinia pestis. Methods It is by optimizing the concentration of Yersinia pestis cells for inoculation,adsorption time and culture temperature,the plaque assay was developed in determination of titer of phage for Yersinia pestis and estimated precision for the developed method. Quality standard of titer was established through identity test and bacterial purity test. Results The optimal cell concentration of Yersinia pestis for inoculation is 7 × 108cells / mL,adsorption is not necessary,and culture temperature is 29 ℃. It shows a good precision by the developed method. Quality standard for titer of phage is no less than 106PFU / mL. Conclusion A plaque assay is developed in determination titer of phage for plaque of Yersinia pestis used in detection of plague vaccine and quality control,and quality standard for titer of phage is established too.
引文
[1]国家药典委员会.中华人民共和国药典(三部)[S].北京:中国医药科技出版社,2010:71-73.
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[7]万学勤,唐冬生,刘爱平,等.幽门螺杆菌毒性噬菌体分离纯化及模拟治疗研究[J].南方医科大学学报,2011,31(2):304-307.
[8]邹艳波,迟秀娟,赵斌.鼠疫菌噬菌体试纸方法的研究[J].中国地方病防治杂志,2008,23(5):331-332.
[9]Anisimov AP,Amoako KK.Treatment of plague:promising alternatives to antibiotics[J].J Med Microbiol,2006,55(Pt 11):1461-1475.
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[11]杜崇涛,王文东,刘军,等.肠出血性大肠杆菌O157:H7噬菌体的分离纯化[J].吉林畜牧兽医,2008,241(29):6-8.
[2]魏东,李恪梅,王国治.用于鼠疫疫苗纯菌检查的噬菌体裂解法的建立[J].国际生物制品学杂志,2013,36(5):237-239.
[3]中国生物制品标准化委员会.中国生物制品规程[S].北京:化学工业出版社,2000:537-538.
[4]吴俐健,张含玉,王笑歌,等.噬菌体生物扩增法检测结核性液体标本的临床应用价值[J].山西医药杂志,2011,40(4):336-338.
[5]黄至澄,王世文,李恪梅,等.不动杆菌属噬菌体分离及其裂解谱分析[J].中华医院感染学杂志,2009,19(3):268-270.
[6]苏胜兵,马红霞,徐凤宇.噬菌体在细菌性疾病诊断和治疗中的应用[J].中国兽医科学,2011,41(5):546-550.
[7]万学勤,唐冬生,刘爱平,等.幽门螺杆菌毒性噬菌体分离纯化及模拟治疗研究[J].南方医科大学学报,2011,31(2):304-307.
[8]邹艳波,迟秀娟,赵斌.鼠疫菌噬菌体试纸方法的研究[J].中国地方病防治杂志,2008,23(5):331-332.
[9]Anisimov AP,Amoako KK.Treatment of plague:promising alternatives to antibiotics[J].J Med Microbiol,2006,55(Pt 11):1461-1475.
[10]武英.应对耐药菌的新方法—噬菌体制剂[J].国外医药抗生素分册,2011,32(5):223-228.
[11]杜崇涛,王文东,刘军,等.肠出血性大肠杆菌O157:H7噬菌体的分离纯化[J].吉林畜牧兽医,2008,241(29):6-8.