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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(P>0.05)。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。
Objective To establish a multiplex PCR system and explore its feasibility for the diagnosis of deep fungal infections. Methods The multiplex PCR system was established by using the common inter-transcriptional region (ITS) of fungi and the specific primers of Aspergillus, Fusarium, Corynebacterium, Candida albicans and Cryptococcus neoformans. Single and quadruple PCR was used to detect simulated body fluid And fungi in 30 clinical specimens of patients with suspected deep fungal infections. Results ITS could be combined with 5 pairs of genus or species-specific primers (duplex PCR), and quadruplex PCR was ITS + Trichosporon + Candida albicans + Cryptococcus neoformans. The positive rates of fungal culture and PCR in 26 cases were 26.67% and 43.33% respectively in the 30 clinical samples. There was no significant difference between the two methods (P> 0.05). Conclusion Single and quadruple PCR have high sensitivity, good stability, simple operation and short time-consuming, which can provide etiological evidence for the early diagnosis of deep fungal infection.