一种用于芽孢杆菌代谢物细胞毒性的快速检测方法

A Rapid Detection Method of the Cytotoxicity of Probiotic Bacillus spp.

为建立一种快速检测芽孢杆菌细胞毒性的方法,通过比较不同的细胞以及活性测定方法,确立以非洲绿猴肾细胞(Vero细胞)为受试细胞,MTT比色法作为芽孢杆菌代谢物细胞毒性的检测方法,并对检测条件进行了优化。利用优化后的方法对81株不同来源芽孢杆菌的细胞毒性进行检测,发现有16株芽孢杆菌具有细胞毒性。通过毒素编码基因PCR、溶血性试验和抗体免疫反应试验进一步验证了16株芽胞杆菌的毒性。与体外测定方法相比,建立的细胞毒性检测方法具有更高的灵敏性,且适用于不同株系芽孢杆菌在细胞水平的安全性评价,具有较高的推广应用价值。

To establish a method for the detection of Bacillus cytotoxicity,the different cells and detection methods were compared. Firstly,African green monkey kidney cell(Vero cells)was as the test cell,MTT colorimetry was selected for detecting the cytotoxicity of Bacillus spp.,and the conditions of cytotoxicity test were optimum. The cytotoxicity of 81 Bacillus strains from different sources was detected using optimum method. The results showed that 16 strains of 81 Bacillus strains were cytotoxic,which were verified by PCR of virulence coden gene detection,hemolysis test and antibody immune response. Compared with those methods in vitro,the Bacillus cytotoxicity detection method established in this study had higher sensitivity and been suitable for the safety evaluation of different strains of Bacillus species at the cell level,which had a high value for popularization and application.