| 화상병원세균 검출을 위한 Conventional PCR 향상 |
최현주1, 김연주1, 최정호1, 최동혁1, 박덕환1,2 
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1강원대학교 스마트농업전공
2강원대학교 생물자원과학부식물의학전공 |
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| Enhancing Conventional PCR for Detection of Erwinia amylovora |
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Hyun Ju Choi1, Yeon Ju Kim1, Jeong Ho Choi1, Dong Hyuk Choi1, Duck Hwan Park1,2
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1Interdisciplinary Program in Smart Agriculture, Kangwon National University, Chuncheon 24341, Korea
2Plant Medicine Program, Division of Bioresource Sciences, College of Agriculture and Life Sciences, Kangwon National University, Chuncheon 24341, Korea |
Correspondence:
Duck Hwan Park, Tel: +82-33-250-6432, Fax: +82-33-259-5558,
Email: dhp@kangwon.ac.kr
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Received: August 23, 2024 Revised: September 16, 2024 Accepted: September 19, 2024 |
| Abstract |
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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. |
| Key Words:
Detection, Erwinia amylovora, Facilitator, Fire blight, Polymerase chain reaction |
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