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Journal of Clinical Microbiology, August 2000, p. 2902-2908, Vol. 38, No. 8
Department of Clinical
Ophthalmology1 and Department of
Pathology,3 The Institute of Ophthalmology and
Moorfields Eye Hospital, London EC1V 9EL, and Department of
Opthalmology, Whippscross Hospital, London E11
1NR,2 United Kingdom
Received 13 January 2000/Returned for modification 8 May
2000/Accepted 3 June 2000
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.
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Rapid Detection and Identification of
Candida, Aspergillus, and Fusarium
Species in Ocular Samples Using Nested PCR


*
Corresponding author. Mailing address: Department of
Clinical Ophthalmology, The Institute of Ophthalmology and Moorfields Eye Hospital, 11-43 Bath St., London EC1V 9EL, United Kingdom. Phone
and Fax: 44-(0)171-608-6931. E-mail:
nokhravi{at}hgmp.mrc.ac.uk.
Present address: Molecular and Population Genetics Laboratory,
Imperial Cancer Research Fund, London WC2A 3PX, United Kingdom.
Present address: Department of Medical Biochemistry, University of
Stellenbosch, Tygerberg 7505, South Africa.
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