Toxicity of a series of tri- and diorganotin(IV) compounds against phytogenic fungal and bacterial lines

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A Simple And Rapid Protocol Of Crude Dna Extraction From Apple Trees For Pcr And Real-Time Pcr Detection Of 'Candidatus Phytoplasma Mali'

none6Different PCR protocols have been established for detection of European fruit trees phytoplasmas; however the majority of the procedures for extracting phytoplasma DNA are complex, time consuming, and expensive, with a risk of contamination or loss of target DNA. In present study, a crude extract preparation method previously used to detect other plant pathogens was adapted to samples from apple trees infected by ‘Candidatus Phytoplasma mali’. End-point and real-time PCR detection of ‘Ca. P. mali’ were used to compare this extraction procedure with an establishedmethod for efficient extraction of purified DNA. The crude extract proved fully adequate for phytoplasma detection in samples from 86 in vitro and 35 in vivo apple shoots or plants and 10 periwinkle plants. High inter- and intra-run reproducibility was obtained for phytoplasma detection with different TaqMan MGB- or SYBR Green-based real-time PCR protocols applied to the crude extracts. Real-time PCR applied to serially diluted crude and purified extracts revealed the same phytoplasma detection limit (dilution up to 105). All results confirm the suitability of this simple, quick, efficient extraction technique for accurate detection of ‘Ca. P. mali’ in different types of apple and periwinkle samplesnoneAldaghi M.; S. Massart; O. Dutrecq; A. Bertaccini; M.H. Jijakli; Ph. LepoivreAldaghi M.; S. Massart; O. Dutrecq; A. Bertaccini; M.H. Jijakli; Ph. Lepoivr

Inter-Laboratory Evaluation Of A Duplex RT-PCR Method Using Crude Extracts For The Simultaneous Detection Of Prune Dwarf Virus And Prunus Necrotic Ringspot Virus

The operational capacity of a duplex RT-PCR method for simultaneous detection of Prune dwarf virus (PDV) and Prunus necrotic ringspot virus (PNRSV) has been established by nine European laboratories. A total of 576 samples from Prunus trees with known sanitary status, corresponding to 32 samples in two repetitions for each laboratory, were analysed. The level of sensitivity achieved by the method was 98.3% for PDV and 90.4% for PNRSV. The specificity was 87.4% for PDV and 94.3% for PNRSV. The unilateral 95% confidence intervals were calculated for all these values. Cohen's Kappa coefficient of repeatability and reproducibility of the technique indicated a strong agreement between data. Likelihood ratios were 7.50 (positive) and 0.02 (negative) for PDV. For PNRSV, the positive likelihood ratio was 15.00 while the negative likelihood ratio was 0.11. In addition, post-test probabilities of infection were calculated to manage the risk associated with the routine use of this method. This allows an accurate test result interpretation to facilitate the integration of this new technique into a certification scheme