| 4种抗原定量方法观察牛支原体培养特征 |
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| 引用本文： | 王展慧,赵萍,陈胜利,郝华芳,秦明明,王绍伟,刘永生,储岳峰.4种抗原定量方法观察牛支原体培养特征[J].中国畜牧兽医,2016,43(10):2634-2639. |
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| 作者姓名： | 王展慧 赵萍 陈胜利 郝华芳 秦明明 王绍伟 刘永生 储岳峰 |
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| 作者单位： | 中国农业科学院兰州兽医研究所, 家畜疫病病原生物学国家重点实验室, 农业部草食动物疫病重点开放实验室, 兰州 730046 |
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| 基金项目： | 国家科技支撑计划（2015BAD12B02）；甘肃省科技支撑计划（1204NKCA071）；兰州市城关区科技计划（2012-2-1） |
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| 摘 要： | 为探索在疫苗研制过程中牛支原体抗原收获时间及抗原定量替代方法，将牛支原体08M株接种于含10%马血清的Thiaucourt's培养基，在110 h内同时监测其颜色变化单位（color change units，CCU）、菌落形成单位（colony forming units，CFU）、菌体蛋白浓度和核酸含量的变化，绘制相应曲线。活菌计数结果（CCU和CFU）显示，牛支原体生长可分为明显的4期，10 h进入对数期，30 h进入稳定期，活菌数最高可达1.0×108 CCU/mL和7.7×107 CFU/mL，75 h进入衰亡期；蛋白浓度从15 h开始迅速增长，至35 h蛋白浓度最高，为72.06 μg/mL，此后维持在58.38~70.65 μg/mL；核酸含量从15 h开始增长，至25 h后Ct值维持在15.32~17.84。结果表明，牛支原体蛋白含量在稳定期初期与活菌数具有良好的相关性。因此，在牛支原体灭活疫苗生产中，稳定期初期是最佳抗原收获时间，可用蛋白浓度法代替活菌计数法进行抗原定量。
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| 关 键 词： | 牛支原体 抗原定量 颜色变化单位 菌落形成单位 蛋白浓度 核酸含量 |
| 收稿时间： | 2016-03-02 |
Observation of the Growth Trends of Mycoplasma bovis with Four Different Antigen Quantitation Methods |
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| WANG Zhan-hui,ZHAO Ping,CHEN Sheng-li,HAO Hua-fang,QIN Ming-ming,WANG Shao-wei,LIU Yong-sheng,CHU Yue-feng.Observation of the Growth Trends of Mycoplasma bovis with Four Different Antigen Quantitation Methods[J].China Animal Husbandry & Veterinary Medicine,2016,43(10):2634-2639. |
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| Authors: | WANG Zhan-hui ZHAO Ping CHEN Sheng-li HAO Hua-fang QIN Ming-ming WANG Shao-wei LIU Yong-sheng CHU Yue-feng |
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| Institution: | Key Laboratory of Epizootic Diseases of Grazing Agricultural Sciences, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China |
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| Abstract: | To explore the antigen harvest time of Mycoplasma bovis (M.bovis) and the antigen quantitation alternative method,M.bovis 08M strain was inoculated in the Thiaucourt's medium.Four growth curve plottings were made by measuring color change units (CCU),colony forming units (CFU),protein concentration and nucleic acid levels.Both the results of CCU and CFU counts showed that the growth of M.bovis was divided into four phases,the logarithmic phase appeared after being cultrured 10 h,the stationary phase appeared at 30 h with the highest number of viable cells up to 1.0×108 CCU/mL and 7.7×107 CFU/mL,and the decline phase started at 75 h.The protein concentration of M.bovis increased rapidly from 15 to 35 h,reached 72.06 μg/mL at 35 h,then maintained at 58.38 to 70.65 μg/mL.The nucleic acid levels of M.bovis increased rapidly from 15 h,and the cycle threshold (Ct) values were maintained between 15.32 to 17.84 after 25 h.These results indicated that there was a good correlation between the protein concentration and viable count of M.bovis at the early stationary phase,which was the best time period to harvest antigen.The protein concentration determination could be an alternative method to quantify antigen contents of M.bovis when preparing inactivated M.bovis vaccine. |
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| Keywords: | Mcoplasma bovis antigen quantitative color change units colony forming units protein concentration nucleic acid contents |
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