[Experimental study of fast diagnosis of mycotic keratitis using molecular biology technical].
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.
Wang
Li-ya
LY
Henan Keratopathy Key Laboratory, Henan Eye Institute, Zhengzhou 450003, China. wangliya55@126.com
Yang
Zi-jian
ZJ
Yang
Xiao-yuan
XY
Sun
Sheng-tao
ST
D015821:Eye Infections, Fungal / Q000175:diagnosis* / Q000382:microbiology; D005658:Fungi / Q000302:isolation & purification*; D006801:Humans; D007634:Keratitis / Q000175:diagnosis* / Q000382:microbiology*; D016533:Mycological Typing Techniques; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity