Enhancing Conventional PCR for Detection of Erwinia amylovora
| Autor: | Hyun Ju Choi, Yeon Ju Kim, Jeong Ho Choi, Dong Hyuk Choi, Duck Hwan Park |
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| Jazyk: | English<br />Korean |
| Rok vydání: | 2024 |
| Předmět: | |
| Zdroj: | Research in Plant Disease, Vol 30, Iss 3, Pp 294-299 (2024) |
| Druh dokumentu: | article |
| ISSN: | 1598-2262 2233-9191 |
| DOI: | 10.5423/RPD.2024.30.3.294 |
| Popis: | Polymerase chain reaction (PCR) methods, including conventional PCR (cPCR) and quantitative real-time PCR (qRT-PCR), with both plasmid- and chromosome-targeting primers, are currently the most reliable methods for detecting Erwinia amylovora due to their high sensitivity and specificity. Despite qRT-PCR's quantitative advantage, cPCR remains an attractive method to detect this bacterium in initial screenings of suspected host plants, as it is cost-effective and does not require skilled personnel in well-equipped laboratories. This study aimed to significantly improve cPCR robustness via application of bovine serum albumin (BSA) as a PCR facilitator, with a modified EaF/R primer pair, as previously reported. Experiments have shown that simple supplementation with BSA (10 mg/ml) enhances cPCR reactions using templates such as genomic DNA, bacterial cells, and infected symptomless host organs, including immature apple fruits and seedlings, with EaF/R primers. The cPCR method described in this study is simple, specific, and reliable, and can be applied in routine assays to diagnose fire blight. |
| Databáze: | Directory of Open Access Journals |
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