Diagnosis of Fusarium keratitis in an animal model using the polymerase chain reaction
Diagnosis of Fusarium keratitis in an animal model using the polymerase chain reaction
复制标题
DOI：
10.1136/bjo.82.3.306
复制
发表时间：
1998-03-01
影响因子：
4.1
通讯作者：
Gloor, P
中科院分区：
文献类型：
--
作者：
Alexandrakis, G;Jalali, S;Gloor, P
目的/背景-本研究的目的是应用聚合酶链反应（PCR）建立一种敏感、特异、快速的检测方法来诊断镰刀菌性角膜炎。镰刀菌是世界上某些地区真菌性角膜感染的最常见原因。这往往是很难建立，角膜炎是由于真菌infection. Methods-镰刀菌solani角膜炎诱导在三只兔子的三只眼睛的真菌悬浮液注射到前角膜基质。在一只兔子中，对侧眼作为对照。从接种后4天到28天，刮取角膜进行培养，然后刮取并擦拭用于PCR分析。用引物直接对镰刀菌角质酶基因的一部分进行PCR，并确定存在或不存在此扩增的靶序列通过琼脂糖凝胶。结果扩增的DNA序列中检测到25的28个样品从感染镰刀菌的角膜，为89%的灵敏度。14例镰刀菌性角膜炎患者中只有3例培养阳性，敏感性为21%。七个控制样品是阴性的PCR为基础的测试，为88%.Conclusion-This PCR为基础的测试-持有承诺，是一种有效的方法诊断镰孢菌角膜炎作为网页镰孢菌感染在其他网站。
Aims/background-The purpose of this study was apply the polymerase chain reaction (PCR) to develop a sensitive, specific, and rapid test to diagnose Fusarium keratitis. Fusarium is the most common cause of fungal corneal infection in some parts of the world. It is often difficult to establish that a keratitis is due to fungal infection.Methods-Fusarium solani keratitis was induced in three eyes of three rabbits by injection of a suspension of the fungus into the anterior corneal stroma. In one rabbit the contralateral eye served as a control. From four to 28 days after inoculation, the corneas were scraped for culture, then scraped and swabbed for PCR analysis. The PCR was performed with primers directed against a portion of the Fusarium cutinase gene, and the presence or absence of this amplified target sequence was determined by agarose gel.Results-The amplified DNA sequence was detected in 25 of 28 samples from the corneas infected with Fusarium, for a sensitivity of 89%. Only three of the 14 samples from these eyes with Fusarium keratitis were positive by culture, for a sensitivity of 21%. Seven of eight control samples were negative by the PCR based test, for a specificity of 88%.Conclusion-This PCR based test-holds promise of being an effective method of diagnosing Fusarium keratitis as web as Fusarium infections at other sites.