用半数变色单位法精确测定支原体活菌滴度

Accurate titration of mycoplasma culture measured by 50% color change unit assay

摘要:

摘要:【目的】使用半数变色单位法实现对支原体活菌滴度的精确测定，以替代当前常用的CCU(变色单位)测定法。【方法】参考病毒滴度TCID50(半数组织细胞感染量)的标准测定方法，对早期学者提出的支原体活菌滴度指标—CCU50(半数变色单位)的测定方法进行改进，并做了重复性、敏感性试验以及与CCU的比对验证试验。为进一步验证该方法的适用性，使用其对猪肺炎支原体和鸡滑液支原体培养物进行了活菌滴度精确测定和评价。【结果】该方法可重复性良好，并具有较强的敏感性和适用性。【结论】CCU50测定法能对支原体活菌滴度进行精确评估，有望成为支原体培养工艺研究、支原体疫苗研发等方面的有效工具。

Abstract:

Abstract:［Objective］ A new method was introduced for precise determination of the live cell titer of mycoplasma culture，and would be a candidate to replace the commonly used CCU (color change unit) assay.［Methods］The CCU50 (50% color change unit) was modified according to the method of TCID50 (50% tissue culture infective dose) assay used for viral titer assessment，and adopted to estimate the live cell titer of mycoplasma． Sensitivity and reproducibility of the CCU50 assay were assessed，and adaptability was checked with M．hyopneumoniae and M．synoviae.［Results］The CCU50 assay showed better reproducibility，sensibility and adaptability than traditional CCU assessment approaches.［Conclusion］The method could be applied to accurate titration determination for mycoplasma，and might be considered as a useful tool for the research of high density fermentation of mycoplasma and development of vaccine.