血液中肺炎链球菌的AFLP快速检测方法的建立

目的 建立扩增片段长度多态性（AFLP）快速检测血液中肺炎链球菌的方法。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。通过对模拟临床血液感染细菌标本的检测，评价该方法的特异性和灵敏度。结果 7个模拟临床血液感染肺炎链球菌标本经AFLP法检测，结果均为阳性；15个模拟临床血液感染非肺炎链球菌标本经该法检测，结果均为阴性；该法检测血液标本中肺炎链球菌的灵敏度为1．5×10_4cfu／mL。结论 建立的AFLP方法检测血液中肺炎链球菌简便快速，特异性好、灵敏度高，适合肺炎链球菌血流感染的早期诊断。

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

Objective To establish a Amplified Fragment Length Polymorphism （AFLP） method for rapid diagnosis of Streptococ, cus pneumonia. Methods Based on the 16S rRNA 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 7 streptococcus pneumonia strains and 15 non-streptococcus pneumonia strains in blood. Results All the 7 strains of streptococcus pneumonia in blood yielded positive results in AFLP, and the 15 strains of other bacteria in blood all showed negative results. The results of streptococcus pneumonia count showed that AFLP was capable of detecting streptococcus pneumonia at a level as low as 1.5 x 104cfu/ml in blood. Conclusion The established AFLP method has high specificity and sensitivity for detecting streptococcus pneumonia in blood and is applicable in field of rapid diagnosis of Streptococcus pneumoniae in blood.