肺炎链球菌的扩增片段长度多态性快速检测方法的建立和应用

目的建立扩增片段长度多态性(AFLP)快速检测肺炎链球菌。方法针对肺炎链球菌的管家基因16SrRNA基因设计特异引物,优化反应条件,建立肺炎链球菌的AFLP检测方法。通过对20株肺炎链球菌和15株非肺炎链球菌进行检测,检验该方法的灵敏度、特异度和实用性。结果活菌计数方法表明建立的AFLP方法检测肺炎链球菌的灵敏度为1.5×103CFU/ml。检测20株肺炎链球菌均为阳性,15株非肺炎链球菌则全部阴性,特异度为100%。结论建立的AFLP方法检测肺炎链球菌灵敏度、特异度高,可用于肺炎链球菌的检测和流行病学调查。

Establishment and Application of AFLP Method for Rapid Diagnosis of Streptococcus Pneumoniae

Objective To establish a amplified fragment length polymorphism(AFLP) method for rapid diagnosis of Streptococcus pneumonia.Methods Based on the 16SrRNA nucleic sequence of streptococcus pneumonia,a pair of primers was designed for AFLP.The reaction conditions were optimized,and the specificity,sensitivity,and practicability of AFLP were tested using 20 streptococcus pneumonia strains and 15 non-streptococcus pneumonia strainss in blood.Results The results of streptococcus pneumonia count showed that AFLP was capable of detecting streptococcus pneumonia at a level as low as 1.5×103CFU/ml.All the 20 strains of streptococcus pneumonia yielded positive results in AFLP,and the 15 strains of other bacteria all showed negative results,with a detection specificity of 100%.Conclusion The established AFLP method has high specificity and sensitivity for detecting streptococcus pneumonia and is applicable in field monitoring and epidemiological study of streptococcus pneumonia.