PURPOSE: To determine the usefulness of polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analysis in the identification and speciation of Candida spp that causes @entity5517 . METHODS: Oligonucleotide primers based on the cytochrome P450 L1 A1 demethylase gene were used to successfully amplify by PCR a single 1.0-kb and a single 500-bp DNA fragment from @entity5132 , @entity9841 , C. krusei, @entity7408 , @entity7409 , and C. pelliculosa genomic DNA. RFLPs within the PCR product were identified after restriction enzyme digestion. RESULTS: The sensitivity of the amplification reaction after two rounds of PCR was 10 fg genomic @entity5132 DNA or one copy of the gene. No amplification product was obtained when DNA from @entity18203 , @entity54 , @entity10437 , @entity1 leukocytes, or 10 species of bacteria was used as a template. Experiments with spiked normal vitreous demonstrated equal sensitivity as long as the volume of vitreous did not exceed 20% of the total PCR volume. RFLP analysis of the PCR product generated from each species obtained from the first- and second-round amplification products enabled species identification after digestion with specific endonucleases. Application of the technique to four clinical samples was successful. CONCLUSIONS: It is expected that the simplicity of the DNA extraction technique allied with the broad specificity of the outer primers for all ophthalmically relevant Candida spp and the sensitivity of the second-round PCR will aid in the detection of fungal DNA in small intraocular samples. PCR-RFLP analysis has great potential in the rapid detection and identification of Candida spp and in the provision of a useful laboratory tool for the future.

Polymerase chain reaction and restriction fragment length polymorphism mediated detection and speciation of Candida spp causing XXXX .

@entity9841 :: ('5482', 'Species') :: ['C. tropicalis'] @entity1 :: ('9606', 'Species') :: ['human'] @entity5517 :: ('MESH:D015817', 'Disease') :: ['ocular infection'] @entity7408 :: ('5478', 'Species') :: ['C. glabrata'] @entity18203 :: ('4929', 'Species') :: ['C. guilliermondii'] @entity10437 :: ('169388', 'Species') :: ['Fusarium solani'] @entity7409 :: ('5480', 'Species') :: ['C. parapsilosis'] @entity5132 :: ('5476', 'Species') :: ['C. albicans'] @entity54 :: ('746128', 'Species') :: ['Aspergillus fumigatus']
@entity5517 :: (MESH:D015817,Disease) :: ['intraocular infection']
