真菌通用引物PCR反应鉴别兔眼真菌性和细菌性角膜溃疡

目的 利用真菌通用引物PCR反应，鉴别兔眼真菌性和细菌性角膜溃疡。方法 根据临床上常见的致病细菌菌株(金葡菌、绿脓菌)，真菌菌株(烟曲霉菌、白念菌、镰刀菌)，制作兔眼角膜溃疡的动物模型，选取一对真菌通用引物，一对寡核苷酸引物B2F(5'ACTITCGTAGGATAG-3')和B4R(5'-TGATCGtcttccataaata-3')，对标本进行聚合酶链反应。结果 在687bp处出现DNA扩增带者为阳性。造模后第3d真菌组出现扩增带阳性的90％，第5d真菌组出现扩增带阳性的96．6％。细菌组始终未出现阳性结果。结论 真菌通用引物PCR反应检测鉴别真菌性与细菌性角膜溃疡有较好的敏感性和特异性。

Detection of Infected Corneal Ulcer in Rabbit Eyes by Fungal General Primer-mediatied Polymerase Chain Reaction

Objective To differentiate fungus corneal ulcer from bacterial ulcer in rabbit eyes by fungal general primer-mediatied polymerase chain reaction(GP-PCR).Methods We set up the infected corneal ulcer model of staphylococcus, pesudomonas aeruginosa, Aspergillus fumigatus, candida albicans and fusarium in rabbit eyes.According to fungal gene orders , we devised and chose out a pair of oligonucleotide primers B2F(5'-ACTTTCGTAGGATAG-3')and B4R(5'-TGATCGTCTTCGATAAATA-3'),whose expanded fraction appeared at 687bp. Results A 687bp specific DNA product was successfully amplified .In models of fungal mycotic keratitis, the sensitivity of PCR were 90.0% at the 3rd day;96.6% at 5th day. There were not specific DNA products to be amplified in bacterium mycotic keratitis.Conclusion The results indicate that GP-PCR is a rapid, sensitive and reliable detection method for fungal corneal ulcer in rabbit eyes.