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KMID : 0364820110470010103
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Korean Journal of Microbiology
2011 Volume.47 No. 1 p.103 ~ p.109
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Development of a Rapid Detection Method for Pectobacterium carotovorum subsp. carotovorum Using the Loop-Mediated Isothermal Amplification (LAMP)
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Kim Jeong-Gu
No Ji-na
Park Dong-suk
Yoon Byoung-su
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Abstract
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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 PCCLAMP. 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°C. Under standard conditions, PCC-LAMP amplified 1×103 copies of clone PCCpBX437 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 arotovorum subsp. carotovorum in the field.
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KEYWORD
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Pectobacterium carotovorum subsp. carotovorum, cabbage, detection, LAMP, soft rot
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