Mycotoxigenic Fungi pp 123-129 | Cite as
Targeting Conserved Genes in Alternaria Species
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Abstract
Real-time polymerase chain reaction (PCR) is a molecular biology technique based on the detection of the fluorescence produced by a reporter molecule, which increases as the reaction proceeds proportionally to the accumulation of the PCR product within each amplification cycle. The fluorescent reporter molecules include dyes that bind to the double-stranded DNA (i.e., SYBR® Green) or sequence-specific probes (i.e., Molecular Beacons or TaqMan® Probes). Real-time PCR provides a tool for accurate and sensitive quantification of target fungal DNA. Here, we describe a TaqMan real-time PCR method for specific detection and quantification of Alternaria spp. The method uses Alternaria-specific primers and probe, targeting the internal transcribed spacer regions ITS1 and ITS2 of the rRNA gene, and a positive amplification control based on 18S rRNA gene.
Key words
TaqMan real-time PCR Internal transcribed spacer Alternaria spp.Notes
Acknowledgments
This work was supported by Grant No. AGL 2006-07659 from the Ministerio de Educación y Ciencia of Spain and the Programa de Vigilancia Sanitaria 2009/AGR-1489 from the Comunidad de Madrid (Spain).
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