[Objective] The paper was to explore the antigen harvest time of Mycoplasrna bovis and the antigen quantification alternative method. [Method] M. bovis 08M strain was inoculated in the Thiaueourt＇s medium containing ...[Objective] The paper was to explore the antigen harvest time of Mycoplasrna bovis and the antigen quantification alternative method. [Method] M. bovis 08M strain was inoculated in the Thiaueourt＇s medium containing 10% horse serum. Four growth curves were plotted by simultaneously measuring color change units （CCU）, colony forming units （CFU）, protein concentration and nucleic acid levels within 110 h. [ Result] The growth of M. bovis was divided into four phases： the longarithmie phase appeared after being cultured for 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 CCU/mL, respectively; and the decline phase started at 75 h. The protein concentration afM. bov/s increased rapidly from 15 to 35 h, reached 72.06 μg/mL at 35 h, then maintained at 53.38 - 70.65 μg/mL. The nucleic acid levels of M. bov/s increased rapidly from 15 h, and the cycle threshold （Ct） values were maintained between 15, 32 and 17.84 after 25 h. [ Conclusion] There was a good correlation between the protein concentration and variable count of M. bov/s 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 content of M. bovis when preparing inactivated M. boviz vaccine. Key words Mycoplasma boyis; Antigen quantification; Color change units; Colony forming units; Protein concentration; Nucleic acid content展开更多
基金Supported by National Key Technology R&D Program(2015BAD12B02)Key Technology R&D Program of Gansu Province(1204NKCA071)Science and Technology Plan of Chengguan District,Lanzhou City(2012-2-1)
文摘[Objective] The paper was to explore the antigen harvest time of Mycoplasrna bovis and the antigen quantification alternative method. [Method] M. bovis 08M strain was inoculated in the Thiaueourt＇s medium containing 10% horse serum. Four growth curves were plotted by simultaneously measuring color change units （CCU）, colony forming units （CFU）, protein concentration and nucleic acid levels within 110 h. [ Result] The growth of M. bovis was divided into four phases： the longarithmie phase appeared after being cultured for 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 CCU/mL, respectively; and the decline phase started at 75 h. The protein concentration afM. bov/s increased rapidly from 15 to 35 h, reached 72.06 μg/mL at 35 h, then maintained at 53.38 - 70.65 μg/mL. The nucleic acid levels of M. bov/s increased rapidly from 15 h, and the cycle threshold （Ct） values were maintained between 15, 32 and 17.84 after 25 h. [ Conclusion] There was a good correlation between the protein concentration and variable count of M. bov/s 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 content of M. bovis when preparing inactivated M. boviz vaccine. Key words Mycoplasma boyis; Antigen quantification; Color change units; Colony forming units; Protein concentration; Nucleic acid content
