肺炎链球菌PCR分型技术的建立与应用
Establishment and application of PCR method in serotyping of Streptococcus pneumonia
摘要目的 建立肺炎链球菌血清型分型的PCR简便方法 ,初步了解肺炎链球菌血清型/群的分布状况.方法 设计合成12种肺炎链球菌血清型/群特异性引物,优化不同血清型/群引物FCR条件,并检测最佳反应浓度、灵敏性以及特异性;初步应用于肺炎链球菌菌株的血清型/群检测.结果引物浓度优化后,12种肺炎链球菌血清型/群特异引物呈现较好的特异性与敏感性;对119株肺炎链球菌菌株进行PCR分型检测,其中113株可分为9个血清型/群(3、5、6A/B、9A/V、14、18、19A、19F、23F),6株未分群.结论 初步建立了12种血清型/群肺炎链球菌PCR分型技术,可用于鉴别人群中主要流行的肺炎链球菌血清型/群.
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abstractsObjective To establish a method based on PCR for serotyping of Streptococcus pneumonia isolates. PCR serotyping method was applied for investigating the serotypes of S. pneumonia strains. Methods 12 pairs of primers targeting different serotypes or S. pneumonia were designed and synthesized. After optimizing the PCR amplification reaction,sensitivity and specificity of each pair was performed. We applied the PCR methods for testing the serotypes of the isolated S. pneumonia strains. Results Each pair of primers showed satisfied PCR sensitivity and specificity. Of all 119 S. pneumonia strains tested by PCR serotyping method,113 isolates were identified (3,5,6A/B,9A/V,14,18,19A,19F,23F) with 6 isolates were unable to be serotyped. Conclusion We deveioped a simple,reliabie and economic method for S. pneumonia serotyping which could be used for testing the serotypes of S. pneumonia that had been prevailed among general population.
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