Quantitative Real-Time PCR Analysis of ‘Candidatus Phytoplasma mali’ Without External Standard Curves

作者: Sanja Baric

DOI: 10.1007/S10341-012-0166-7

关键词: Computational biologyMalusBiologyPhytoplasmaBotanyAbsolute quantificationCandidatus Phytoplasma maliPathogenTaqMan16S ribosomal RNAStandard curve

摘要: Apple proliferation caused by ‘Candidatus Phytoplasma mali’ is an economically important disease of apple (Malus ´ domestica). The availability a simple quantitative approach to assess pathogen load in infected host plants would certainly contribute better understanding pathogenesis and epidemiology. This study proposes quantification not requiring the analysis external standard curves. It based on simultaneous detection 16S rRNA gene 1-aminocyclopropane-1-carboxylate oxidase plant single-tube reaction using TaqMan chemistry. quantity phytoplasma relative its determined as difference between CT values two target genes (ΔCT). To agreement with curve-based method, dataset 450 DNA samples from trees was reanalysed. Comparison ΔCT-based quantities corresponding absolute revealed high degrees methods. ΔCT procedure can thus be considered adequate for tissue. In addition, this ensures improved methodological standardisation increased throughput.

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