Res. Plant Dis > Volume 30(3); 2024 > Article
Research in Plant Disease 2024;30(3):294-299.
DOI: https://doi.org/10.5423/RPD.2024.30.3.294    Published online September 30, 2024.
화상병원세균 검출을 위한 Conventional PCR 향상
최현주1, 김연주1, 최정호1, 최동혁1, 박덕환1,2 
1강원대학교 스마트농업전공
2강원대학교 생물자원과학부식물의학전공
 
Enhancing Conventional PCR for Detection of Erwinia amylovora
Hyun Ju Choi1, Yeon Ju Kim1, Jeong Ho Choi1, Dong Hyuk Choi1, Duck Hwan Park1,2 
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
Received: August 23, 2024   Revised: September 16, 2024   Accepted: September 19, 2024
Abstract
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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