真菌四重PCR体系的建立及其临床应用

发布时间：2018-01-02 13:20

本文关键词：真菌四重PCR体系的建立及其临床应用　出处：《中国皮肤性病学杂志》2015年05期 　论文类型：期刊论文

【摘要】：目的建立多重PCR体系并探讨其用于深部真菌感染诊断的可行性。方法用真菌通用引物内转录间区(ITS)联合曲霉属、镰刀菌属、毛孢子菌属、白念珠菌、新生隐球菌特异性引物建立多重PCR体系,用单重和四重PCR检测模拟体液和30例可疑深部真菌感染患者临床标本中的真菌。结果ITS可与5对属或种特异性引物任意组合(二重PCR),四重PCR组合为ITS+毛孢子菌属+白念珠菌+新生隐球菌。30例临床标本中,真菌培养、PCR检测的阳性率分别为26.67%和43.33%,两种方法的检出率差异无统计学意义(P0.05)。结论单重和四重PCR敏感性高、稳定性好、操作简单、耗时短,可为深部真菌感染的早期诊断提供病原学依据。
[Abstract]:Objective to establish a multiplex PCR system and discusses its feasibility for diagnosis of deep fungal infection. Methods using fungal universal primers internal transcribed spacer (ITS) combined with Aspergillus, Fusarium, Trichosporon, Candida, Cryptococcus neoformans specific primers to establish the multiplex PCR system, simulated body fluid and clinical specimens in patients with fungal 30 cases of suspected deep fungal infection with single and four PCR detection. Results ITS and 5 of genera or species specific primers combinations (double PCR), four PCR combination ITS+ + Trichosporon albicans and Cryptococcus neoformans.30 cases in clinical specimens, fungal culture positive rate of PCR detection were 26.67% and 43.33%, the two methods of detection rates were not statistically significant (P0.05). Conclusion the single and four PCR with high sensitivity, good stability, simple operation, short time, can provide the basis for early diagnosis of deep fungal infection of the pathogen.

【作者单位】：广东医学院附属医院皮肤科;广东医学院附属医院重症医学科;
【基金】：湛江市财政资金科技专项竞争性分配项目(湛科[2010]174号);湛江市财政资金科技专项竞争性分配项目(2012C0302-46)
【分类号】：R756
【正文快照】： 真菌培养和组织病理检查是深部真菌感染诊断的金标准,但存在耗时长、敏感性较低、取材困难等缺点,不能满足早期诊断的要求[1]。PCR法具有特异性和敏感性高、检测快速的优点,已广泛用于真菌鉴定、致病性、药物作用机制、耐药性等方面研究[2]。多重PCR利用多对特异性引物同时扩

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