[应用分子生物学技术快速诊断真菌性角膜炎的实验研究]
[Experimental study of fast diagnosis of mycotic keratitis using molecular biology technical].
作者信息
Wang Li-ya, Yang Zi-jian, Yang Xiao-yuan, Sun Sheng-tao
机构信息
Henan Keratopathy Key Laboratory, Henan Eye Institute, Zhengzhou 450003, China.
出版信息
Zhonghua Yan Ke Za Zhi. 2007 Mar;43(3):256-9.
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, Fusarium equiseti, Fusarium graminum, Aspergillus fumigatus, Aspergillus flavus, Aspergillus niger, Penicillium 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, Penicillium 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.
目的
建立一种使用多重聚合酶链反应（multi-PCR）系统快速诊断真菌性角膜炎的方法。
方法
通过多重聚合酶链反应系统检测9种重要的医学真菌（茄病镰刀菌、串珠镰刀菌、木贼镰刀菌、禾谷镰刀菌、烟曲霉、黄曲霉、黑曲霉、隐球菌和新月弯孢霉）。该方法用于鉴定临床培养分离株和临床样本。
结果
在该多重聚合酶链反应系统中，镰刀菌属扩增出一条360bp的片段，曲霉属扩增出一条430bp的片段，隐球菌属扩增出一条245bp的片段，新月弯孢霉扩增出一条300bp的片段。人DNA以及人眼感染中其他微生物的DNA在该多重聚合酶链反应系统中均获得阴性结果。该多重聚合酶链反应系统的灵敏度为10fg。
结论
多重聚合酶链反应系统可检测和鉴定重要的临床真菌。多重聚合酶链反应方法被证明是一种快速、灵敏且特异的技术，在临床应用中具有良好的前景。
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