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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。
The gyr B gene was selected as the target gene to design a specific primer for PCR detection of Serratia. The fragment of the specific primer amplified product has a size of 279 bp. The 14 selected strains of Serratia in the study represent 7 different species of Serratia. In addition, two Salmonella species, two Klebsiella species, and five other enteric bacteria were used for primer-specific validation. The results showed that the size of amplified bands was the same as that of the expected amplified products, while the other 9 non-Serratia species did not have amplified bands. The detection limit of Serratia marcescens was 9.785 pg . Four isolates of Serratia marcescens isolated from bees and three isolates of Enterococcus faecium isolated from imported fishmeal were tested for positive results. Studies have shown that the PCR method based on the gyr B gene has the characteristics of strong specificity, fastness and high sensitivity in the detection of Serratia.