环保微生物菌剂常用5种芽胞杆菌的PCR鉴定

枯草芽胞杆菌（Bacillus subtilis）、地表芽胞杆菌（Bacillus licheniformis）、巨大芽胞杆菌（Bacillus megate．rium）、短小芽胞杆菌（Bacillus pumilus）、环状芽胞杆菌（Bacillu scirculans）是5种环保微生物菌剂中常用的芽胞杆菌，我国每年有大量此类菌剂进口。针对这些菌的传统理化检测方法费时费力，检测部门急需建立一种快速、准确的检测方法。利用GenBank上已有gyrB、rpoB、gdh基因序列分别设计和筛选上述菌种的特异性引物，经优化建立PCR检测方法。结果显示，使用所设计的特异引物，每种茼均扩增得到特异单1条带，能准确鉴定上述菌种并且与其他对照芽胞杆菌及非芽胞杆菌无交叉反应，送检样品的PCR检测结果与常规方法检测结果一致。建立的特异PCR方法具有良好的重复性和准确性，可快速鉴定上述菌种，具有很好的推广应用前景。

The Identification of Five Bacillus Species Used in Environment Microbe Agentia with PCR Method

The five Bacillus species, B. subtilis, B. licheniformis, B. megaterium, B. pumilus and B. circulans are generally used as environment microbe agentia, which are imported heavy every year. Because the traditional detecting methods for these strains arc complicated and time-consuming, the new rapid and accurate identification method is needed to set up. By analyzing nucleotide sequences of the rpoB,gyrB and gdh genes from the GanBank, the speciesspecific PCR primers were designed and the reaction conditions were optimized in this research. The result showed that all five species generated a specific amplification strand while the other contrast strains were no cross reaction with the primers. Detecting consequences of submitted samples were uniformity between the PCR and traditional chemical methods. The specific PCR method established in this study has good repeatability and accuracy, and maybe a potential method for the application in rapid identification of these Bacillus species.