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| Content Provider | Springer Nature Link |
|---|---|
| Author | Bellis, Palmira Schena, Leonardo Cariddi, Corrado |
| Copyright Year | 2007 |
| Abstract | A specific primer couple (E3–E4) amplifying a single DNA fragment of 111 bp from plasmid pEA29 was designed to identify, detect and quantify Erwinia amylovora by real-time Scorpion-PCR. Specificity of primers and probe was assessed both by means of BLAST analyses and by using genomic DNA from a large number of E. amylovora isolates and other bacteria. In Scorpion-PCR, the limit of detection was of 1 pg of total DNA and a high correlation (r = 0.999) was achieved between target DNA quantity and cycle threshold (Ct). Combining two sequential amplifications with conventional reported primers (PEANT1–PEANT2) and Scorpion primers (E3 Scorpion-E4) the detection limit was of 1 fg (nested Scorpion-PCR). Using serial dilution of the bacterial suspensions the limit of detection was 3.2 × 104 CFU ml−1 in Scorpion-PCR and 2.8 × 102 CFU ml−1 in nested Scorpion-PCR. Real-time PCR combined with effective procedures for DNA extraction enabled the detection and the quantification of the epiphytic population of E. amylovora in the washings of flowers and leaves of artificially inoculated pear. A significant correlation (r = 0.92) was achieved between pathogen CFU on semi-selective media and the corresponding target DNA concentration evaluated by real-time PCR. |
| Starting Page | 11 |
| Ending Page | 22 |
| Page Count | 12 |
| File Format | |
| ISSN | 09291873 |
| Journal | European Journal of Plant Pathology |
| Volume Number | 118 |
| Issue Number | 1 |
| e-ISSN | 15738469 |
| Language | English |
| Publisher | Kluwer Academic Publishers |
| Publisher Date | 2007-03-29 |
| Publisher Place | Dordrecht |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Fire Blight Molecular detection Nested PCR Plasmid pEA29 Scorpion-PCR (duplex format) Ecology Plant Sciences Plant Pathology |
| Content Type | Text |
| Resource Type | Article |
| Subject | Plant Science Horticulture Agronomy and Crop Science |
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