Campus Units

Food Science and Human Nutrition

Document Type

Article

Publication Version

Published Version

Publication Date

11-26-2014

Journal or Book Title

journal of Clinical Microbiology

Volume

53

Issue

2

First Page

511

Last Page

521

DOI

10.1128/JCM.02417-14

Abstract

Candida albicans is an important cause of systemic fungal infections, and rapid diagnostics for identifying and differentiating C. albicans from other Candida species are critical for the timely application of appropriate antimicrobial therapy, improved patient outcomes, and pharmaceutical cost savings. In this work, two 28S rRNA-directed peptide nucleic acid-fluorescence in situ hybridization (PNA-FISH) probes, P-Ca726 (targeting a novel region of the ribosome) and P-CalB2208 (targeting a previously reported region), were evaluated. Hybridization conditions were optimized by using both fluorescence microscopy (FM) and flow cytometry (FCM), and probes were screened for specificity and discriminative ability against a panel of C. albicans and various nontarget Candida spp. The performance of these PNA probes was compared quantitatively against that of DNA probes or DNA probe/helper combinations directed against the same target regions. Ratiometric analyses of FCM results indicated that both the hybridization quality and yield of the PNA probes were higher than those of the DNA probes. In FCM-based comparisons of the PNA probes, P-Ca726 was found to be highly specific, showing 2.5- to 5.5-fold-higher discriminatory power for C. albicans than P-CalB2208. The use of formamide further improved the performance of the new probe. Our results reinforce the significant practical and diagnostic advantages of PNA probes over their DNA counterparts for FISH and indicate that P-Ca726 may be used advantageously for the rapid and specific identification of C. albicans in clinical and related applications, especially when combined with FCM.

Comments

This article is published as Kim, H.J. and Brehm-Stecher, B.F. Design and evaluation of peptide nucleic acid probes for specific identification of Candida albicans. J. Clin. Microbiol. 53: 511-21 (2015). Doi: 10.1128/JCM.02417-14. Posted with permission.

Copyright Owner

American Society for Microbiology

Language

en

File Format

application/pdf

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