Oligonucleotide Microarray for the Identification of Potential Mycotoxigenic Fungi

Overview

Journal BMC Microbiol

Publisher Biomed Central

Specialty Microbiology

Date 2010 Mar 24

PMID 20307326

Citations 3

Authors

Sabine Lezar

Eugenia Barros

Affiliations

Soon will be listed here.

Abstract

Background: Mycotoxins are secondary metabolites which are produced by numerous fungi and pose a continuous challenge to the safety and quality of food commodities in South Africa. These toxins have toxicologically relevant effects on humans and animals that eat contaminated foods. In this study, a diagnostic DNA microarray was developed for the identification of the most common food-borne fungi, as well as the genes leading to toxin production.

Results: A total of 40 potentially mycotoxigenic fungi isolated from different food commodities, as well as the genes that are involved in the mycotoxin synthetic pathways, were analyzed. For fungal identification, oligonucleotide probes were designed by exploiting the sequence variations of the elongation factor 1-alpha (EF-1 alpha) coding regions and the internal transcribed spacer (ITS) regions of the rRNA gene cassette. For the detection of fungi able to produce mycotoxins, oligonucleotide probes directed towards genes leading to toxin production from different fungal strains were identified in data available in the public domain. The probes selected for fungal identification and the probes specific for toxin producing genes were spotted onto microarray slides.

Conclusions: The diagnostic microarray developed can be used to identify single pure strains or cultures of potentially mycotoxigenic fungi as well as genes leading to toxin production in both laboratory samples and maize-derived foods offering an interesting potential for microbiological laboratories.

Citing Articles

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References

Kristensen R, Gauthier G, Berdal K, Hamels S, Remacle J, Holst-Jensen A . DNA microarray to detect and identify trichothecene- and moniliformin-producing Fusarium species. J Appl Microbiol. 2007; 102(4):1060-70. DOI: 10.1111/j.1365-2672.2006.03165.x. View

Yoder W, Christianson L . Species-specific primers resolve members of Fusarium section Fusarium. Taxonomic status of the edible "Quorn" fungus reevaluated. Fungal Genet Biol. 1998; 23(1):68-80. DOI: 10.1006/fgbi.1997.1027. View

DeSantis T, Stone C, Murray S, Moberg J, Andersen G . Rapid quantification and taxonomic classification of environmental DNA from both prokaryotic and eukaryotic origins using a microarray. FEMS Microbiol Lett. 2005; 245(2):271-8. DOI: 10.1016/j.femsle.2005.03.016. View

Niessen M, Vogel R . Group specific PCR-detection of potential trichothecene-producing Fusarium-species in pure cultures and cereal samples. Syst Appl Microbiol. 1999; 21(4):618-31. DOI: 10.1016/S0723-2020(98)80075-1. View

Bluhm B, Flaherty J, Cousin M, Woloshuk C . Multiplex polymerase chain reaction assay for the differential detection of trichothecene- and fumonisin-producing species of Fusarium in cornmeal. J Food Prot. 2002; 65(12):1955-61. DOI: 10.4315/0362-028x-65.12.1955. View

Vora G, Meador C, Stenger D, Andreadis J . Nucleic acid amplification strategies for DNA microarray-based pathogen detection. Appl Environ Microbiol. 2004; 70(5):3047-54. PMC: 404398. DOI: 10.1128/AEM.70.5.3047-3054.2004. View

Zeng X, Kong F, Halliday C, Chen S, Lau A, Playford G . Reverse line blot hybridization assay for identification of medically important fungi from culture and clinical specimens. J Clin Microbiol. 2007; 45(9):2872-80. PMC: 2045312. DOI: 10.1128/JCM.00687-07. View

Lee T, Han Y, Kim K, Yun S, Lee Y . Tri13 and Tri7 determine deoxynivalenol- and nivalenol-producing chemotypes of Gibberella zeae. Appl Environ Microbiol. 2002; 68(5):2148-54. PMC: 127587. DOI: 10.1128/AEM.68.5.2148-2154.2002. View

Martin K, Rygiewicz P . Fungal-specific PCR primers developed for analysis of the ITS region of environmental DNA extracts. BMC Microbiol. 2005; 5:28. PMC: 1156903. DOI: 10.1186/1471-2180-5-28. View

10.

Niessen L . PCR-based diagnosis and quantification of mycotoxin producing fungi. Int J Food Microbiol. 2007; 119(1-2):38-46. DOI: 10.1016/j.ijfoodmicro.2007.07.023. View

11.

Mayer Z, Bagnara A, Farber P, Geisen R . Quantification of the copy number of nor-1, a gene of the aflatoxin biosynthetic pathway by real-time PCR, and its correlation to the cfu of Aspergillus flavus in foods. Int J Food Microbiol. 2003; 82(2):143-51. DOI: 10.1016/s0168-1605(02)00250-7. View

12.

Kane M, Jatkoe T, Stumpf C, Lu J, Thomas J, Madore S . Assessment of the sensitivity and specificity of oligonucleotide (50mer) microarrays. Nucleic Acids Res. 2000; 28(22):4552-7. PMC: 113865. DOI: 10.1093/nar/28.22.4552. View

13.

Graf A, Gasser B, Dragosits M, Sauer M, Leparc G, Tuchler T . Novel insights into the unfolded protein response using Pichia pastoris specific DNA microarrays. BMC Genomics. 2008; 9:390. PMC: 2533675. DOI: 10.1186/1471-2164-9-390. View

14.

Dombrink-Kurtzman M . The sequence of the isoepoxydon dehydrogenase gene of the patulin biosynthetic pathway in Penicillium species. Antonie Van Leeuwenhoek. 2006; 91(2):179-89. DOI: 10.1007/s10482-006-9109-3. View

15.

Anthony R, Brown T, French G . Rapid diagnosis of bacteremia by universal amplification of 23S ribosomal DNA followed by hybridization to an oligonucleotide array. J Clin Microbiol. 2000; 38(2):781-8. PMC: 86203. DOI: 10.1128/JCM.38.2.781-788.2000. View

16.

Volokhov D, Rasooly A, Chumakov K, Chizhikov V . Identification of Listeria species by microarray-based assay. J Clin Microbiol. 2002; 40(12):4720-8. PMC: 154633. DOI: 10.1128/JCM.40.12.4720-4728.2002. View

17.

Lane S, Evermann J, Loge F, Call D . Amplicon secondary structure prevents target hybridization to oligonucleotide microarrays. Biosens Bioelectron. 2004; 20(4):728-35. DOI: 10.1016/j.bios.2004.04.014. View

18.

You Y, Moreira B, Behlke M, Owczarzy R . Design of LNA probes that improve mismatch discrimination. Nucleic Acids Res. 2006; 34(8):e60. PMC: 1456327. DOI: 10.1093/nar/gkl175. View

19.

Leinberger D, Schumacher U, Autenrieth I, Bachmann T . Development of a DNA microarray for detection and identification of fungal pathogens involved in invasive mycoses. J Clin Microbiol. 2005; 43(10):4943-53. PMC: 1248467. DOI: 10.1128/JCM.43.10.4943-4953.2005. View

20.

Letowski J, Brousseau R, Masson L . Designing better probes: effect of probe size, mismatch position and number on hybridization in DNA oligonucleotide microarrays. J Microbiol Methods. 2004; 57(2):269-78. DOI: 10.1016/j.mimet.2004.02.002. View