基于gyrB基因建立快速检测沙雷菌的PCR方法

Rapid detection of Serratia spp. by PCR using the primers targeting the gyrB gene

选择gyr B基因作为靶基因设计特异性引物用于沙雷菌属的PCR检测,该特异性引物扩增产物的片段大小为279 bp。研究所选用的14株沙雷菌分别代表沙雷菌属的7个不同种。此外,2种沙门菌、2种克雷伯菌以及其他5种肠道菌用于进行引物特异性的验证。结果表明:扩增的条带与预期的扩增产物片段大小一致,而非沙雷菌属的其他9个种属均没有扩增条带;该方法检测沙雷菌的最低检测限为9.785 pg。通过对4株从蜜蜂中分离所得的黏质沙雷菌和3株从进口鱼粉中分离所得的居泉沙雷菌进行检测,结果均为阳性。研究表明,基于gyr B基因建立的PCR方法在沙雷菌属的检测中具有特异性强、快速和灵敏度高等特点。

A species-specific primer pair was designed for the PCR detection of Serratia spp. using gyr B gene as the target gene with the fragment size of 279 bp. Fourteen members of the Serratia family（ representing seven Serratia species） were chosen to verify the specificity of the primers. Additionally,two species from Salmonella,two from Klebsiella,and five other species belonging to five other genera of Enterobacteriaceae,were tested for primer cross-reaction,and all the tested strains gave negative results. The limit of detection for Serratia genomic DNA using the gyr B gene was 9. 785 pg per PCR reaction. Four clinical samples from bees and three clinical samples from imported fish meal were detected,and all clinical samples showed positive results. This PCR assay provides a specific,rapid,and sensitive method to detect Serratia species based on their gyr B gene.