采用通用引物PCR配合SSCP及RFLP技术快速检测常见病原菌

Rapid identification of pathogens using UPPCR with SSCP and RFLP analysis

目的 建立一种快速检测病原菌的方法以利于临床诊断。方法 选取 10种具有代表性的病原菌 ,采用通用引物PCR(UPPCR)对其 16SrRNA基因进行扩增 ,并对PCR产物进行限制性酶切片段长度多态性分析 (RFLP)和单链构象多态性分析 (SSCP)。结果 RFLP电泳图谱呈现多态性 ,但半数菌的图谱两两相同或相似 ;SSCP电泳图谱各异 ,可以相互区分。结论 UPPCR

Objective To establish a method for the identification of pathogens for fast diagnosis. Methods By universal primer polymerase chain reaction (UPPCR), the 16S ribosomal RNA genes of ten representative pathogens were amplified. Then the PCR products were analyzed by single strand configuration polymorphism (SSCP) and restrict fragment length polymorphism (RFLP). Results RFLP map expressed polymorphism, but there were either the same or similar figures between two or among three pathogens in half of test strains. However, there were significantly different bands in all of the pathogens by SSCP. Conclusion UPPCR SSCP analysis can rapidly and conveniently identify bacterial pathogens. [