多重PCR快速诊断单纯疱疹病毒、真菌及棘阿米巴角膜炎

发布时间：2018-04-24 07:23

本文选题：单纯疱疹病毒性角膜炎 + 真菌性角膜炎　；参考：《广东医学》2014年15期

【摘要】：目的建立多重PCR体系对单纯疱疹病毒(HSV)、真菌及棘阿米巴性角膜炎同时诊断,并对真菌性角膜炎主要致病真菌进行菌属鉴定。方法建立两个多重PCR体系,检测HSVⅠ型、HSVⅡ型、4株棘阿米巴分离株及9种主要真菌性角膜炎致病真菌,比较该体系对真菌临床菌株、人类基因组及其他血液常见致病微生物的检测效果。结果根据DNA模板,该体系可扩增出不同特异性条带,能够准确检测HSV和棘阿米巴,并将9种待测真菌分为4个菌属。84株真菌受试菌株中81株的鉴定结果与常规鉴定结果一致。该体系对人类基因组及其他血液常见致病微生物DNA的扩增结果均为阴性。模板最低阳性扩增的浓度均为10 fg。结论通过两个多重PCR体系可同时诊断HSV、真菌及棘阿米巴性角膜炎,并可将真菌性角膜炎在菌属水平进行鉴定。该方法具有高效、简便、特异、灵敏的特点,具有较好的临床应用前景。
[Abstract]:Objective to establish a multiplex PCR system for simultaneous diagnosis of herpes simplex virus (HSV), fungi and Acanthamoeba keratitis, and to identify the main pathogenic fungi of fungal keratitis. Methods two multiplex PCR systems were established to detect 4 strains of Acanthoamoeba and 9 major fungal keratitis pathogenic fungi. Detection of common pathogenic microorganisms in human genome and other blood. Results according to DNA template, different specific bands could be amplified by this system, and HSV and Acanthamoeba could be detected accurately. The identification results of 81 strains of fungi tested in 4 genera and 84 strains were consistent with those of routine identification. The results of DNA amplification of human genome and other common pathogenic microorganisms in human blood were negative. The lowest positive amplification concentration of template was 10 fg. Conclusion it is possible to diagnose HSV, fungi and Acanthamoeba keratitis at the same time by using two multiple PCR systems, and to identify fungal keratitis at the genus level. The method has the characteristics of high efficiency, simplicity, specificity and sensitivity, and has a good prospect of clinical application.
【作者单位】：安徽理工大学医学院免疫与检验教研室免疫与感染研究所;安徽省淮南市朝阳医院检验科;淮南东方医院集团总医院检验科;
【基金】：国家级大学生创新创业训练计划项目(编号:201210361112)
【分类号】：R772.21

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