Affiliation:
1. Department of Clinical Ophthalmology1 and
2. and Department of Opthalmology, Whippscross Hospital, London E11 1NR,2 United Kingdom
3. Department of Pathology,3 The Institute of Ophthalmology and Moorfields Eye Hospital, London EC1V 9EL,
Abstract
ABSTRACT
A protocol for the rapid detection of fungal DNA in ocular samples, derived from three species,
Candida albicans
,
Aspergillus fumigatus
, and
Fusarium solani
, has been developed. Two novel panfungal primers complementary to 18S rRNA sequences present in all three species were designed. Panfungal PCR was followed by three nested PCRs utilizing species-specific primers. PCR sensitivity ranged from 50 to 100 fg of free DNA and between one and two
C. albicans
organisms. In addition, we also developed a rapid and reliable DNA extraction protocol. This protocol minimized DNA loss during extraction, whilst removing compounds from vitreous and aqueous fluids that have previously been shown to have inhibitory effects on PCR. Preliminary results obtained after testing the protocol on three patient samples support culture results and medical history. However, one patient was PCR positive but culture negative, suggesting that the sensitivity of this protocol may exceed that of traditional culture techniques. This system, therefore, constitutes an additional protocol that may significantly aid patient management in cases where fungal endophthalmitis is suspected.
Publisher
American Society for Microbiology
Cited by
140 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献