多重聚合酶链反应对真菌性角膜炎快速诊断与菌属鉴定的实验研究  

Experimental study of fast diagnosis of mycotic keratitis using molecular biology technical

作　　者：王丽娅[1] 杨子建[1] 杨潇远[1] 孙声桃[1] 

机构地区：[1]河南省眼科研究所 河南省角膜病重点实验室,郑州450003

出　　处：《中华眼科杂志》2007年第3期256-259,共4页Chinese Journal of Ophthalmology

基　　金：河南省杰出人才创新基金项目（0321002000）;河南省医学科技创新人才工程计划项目（20001102）

摘　　要：目的探讨多重聚合酶链反应（PCR）对真菌性角膜炎快速诊断并同时鉴定致病菌类型的临床应用。方法通过建立多重PCR体系，对腐皮镰孢菌、禾谷镰孢菌、木贼镰孢菌、串珠镰孢菌、烟曲霉菌、黄曲霉菌、黑曲霉菌、新月弯孢菌及牵连青霉菌标准菌株的DNA进行检测，并将此方法应用于临床分离菌株和临床标本的检测。结果在本多重PCR体系中，镰孢菌属、曲霉菌属、牵连青霉菌及新月弯孢菌DNA的扩增条带长度分别为360、430、245及300bp。人基因组DNA及其他眼部常见致病微生物DNA未扩增出条带，表明多重PCR体系具有高度的特异性。多重PCR体系的敏感性为10fg。结论多重PCR体系可对临床常见角膜致病真菌快速诊断并鉴定到菌属，特异性强，敏感度高。Objective To establish a method for fast diagnosis of mycotic keratitis using multi-PCR system. Methods Detecting 9 important medical fungi species （ Fusarium solani, Fusarium moniliform, Fusariume equiseti, Fusarium graminum, Aspergillus fumigatus, Aspergillus flavus, Aspergillus niger, Penicillum implicatum and Curvularia lunata）through a multi-PCR system. This method was used to identify the clinical cultured isolates and clinical samples. Results In this multi-PCR system, Fusarium species amplified a fragment of 360 bp, Aspergillus species amplified a fragment of 430 bp, Penicillum implicatum species amplified a fragment of 245 bp, and Curvularia lunata species amplified a fragment of 300 bp. Human DNA and DNA of other microorganism in human ocular infection obtained negative results in this multi-PCR system. The sensitivity of the multi-PCR system was 10 fg. Conclusions Important clinical fungi could be detected and identified by the multi-PCR system. The multi-PCR method was proved to be a fast, sensitive and specific technology and has a good prospect in clinical application.

关 键 词：眼感染 真菌性 角膜炎 聚合酶链反应 

分 类 号：R686[医药卫生—骨科学]