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.
Copyright Owner
American Society for Microbiology
Copyright Date
2015
Language
en
File Format
application/pdf
Recommended Citation
Kim, H. J. and Brehm-Stecher, Byron F., "Design and evaluation of peptide nucleic acid probes for specific identification of Candida albicans." (2014). Food Science and Human Nutrition Publications. 174.
https://lib.dr.iastate.edu/fshn_ag_pubs/174
Included in
Food Microbiology Commons, Fresh Water Studies Commons, Human and Clinical Nutrition Commons, Molecular, Genetic, and Biochemical Nutrition Commons, Plant Sciences Commons
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.