23S rRNA通用引物的设计及在气性坏疽快速检测中的应用

Design and application of 23S rRNA general primers in fast detection of gas gangrene

目的设计23S rRNA通用引物并将其用于气性坏疽快速检测基因芯片。方法 (1)设计23S rRNA通用引物,并对气性坏疽的主要致病菌进行聚合酶链反应(PCR)扩增。(2)对外伤感染中常见其他微生物进行检测。(3)测定该通用引物PCR的灵敏度。结果 (1)设计出1对23S rRNA通用引物,该通用引物对气性坏疽主要病原菌都能进行扩增。(2)该通用引物PCR对外伤感染中常见非梭菌微生物都不扩增。(3)该通用引物PCR的最高灵敏度为1×103cfu/mL(产气荚膜梭菌)。结论设计出了1对23S rRNA通用引物,能够满足气性坏疽快速检测基因芯片的要求。

Objective To design and application of 23S rRNA general primers in the fast gene chip detection of gas gangrene. Methods （1）23S rRNA general primers were designed and used to amplify target genes of the main pathogenic bacteria of gas gan grene. （2）Other common microbes in external injury infection were deteced. （3）The sensitivity of PCR using designed general prim- ers was analyzed. Results （1）A pair of 23S rRNA general primers were designed and target genes of the main pathogenic bacteria of gas gangrene could be amplified by using the designed general primers. （2）Other common microbes in external injury infection could not be detected by using same primers. （3）The highest sensitivity of PCR using designed primers was 1 ×10^3 cfu/mL（C, perfringens）. Conclusion A pair of 23S rRNA general primers,designed in this investigation,can be used to construct the gene chip for fast detection of gas gangrene.