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Development of a Rapid Detection Method for Pectobacterium carotovorum subsp. carotovorum Using the Loop-Mediated Isothermal Amplification (LAMP)  

Kim, Jeong-Gu (Genomics Division, National Academy of Agricultural Science, Rural Development Administration)
No, Ji-Na (Department of Life Science, College of Natural Science, Kyonggi University)
Park, Dong-Suk (Genomics Division, National Academy of Agricultural Science, Rural Development Administration)
Yoon, Byoung-Su (Department of Life Science, College of Natural Science, Kyonggi University)
Publication Information
Korean Journal of Microbiology / v.47, no.2, 2011 , pp. 103-109 More about this Journal
Abstract
Pectobacterium carotovorum subsp. carotovorum is the causative agent of soft rot in crops such as potato and cabbages. Loop-mediated isothermal amplification (LAMP) is a simple DNA amplification method, as well as isothermal PCR technique. In this study, a new method for the rapid detection of Pectobacterium carotovorum subsp. carotovorum was developed using LAMP that named PCC-LAMP. Based on lytic murein transglycolase gene of Pectobacterium carotovorum subsp. carotovorum, a set of four primers for LAMP was designed. The optimal PCC-LAMP reaction temperature was established at $61^{\circ}C$. Under standard conditions, PCC-LAMP amplified $1{\times}10^3$ copies of clone PCC-pBX437 per reaction. Further, this method can also assay directly by SYBR Green I without electrophoresis. Amplification was not detected for five other bacterial species. In conclusion, PCC-LAMP may be a useful method for the detection Pectobacterium carotovorum subsp. carotovorum in the field.
Keywords
Pectobacterium carotovorum subsp. carotovorum; cabbage; detection; LAMP; soft rot;
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