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KMID : 0386320190520010029
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Korean Leprosy Bulletin
2019 Volume.52 No. 1 p.29 ~ p.40
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Comparison of Effective DNA Extraction method for Molecular biological study of Mycobacterium leprae
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Kim Yun-Ji
Park Jin-Ho
Kim Jong-Pill
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Abstract
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Background: Leprosy is an important health problem in many geographical areas yet. It is caused through a cough or contact with fluid from the nose of a person infected by Mycobacterium leprae. Study of DNA from M. leprae is important to understand essentiality for leprosy. However, there is no standard in many parts, so various studies are needed.
Objects: In this study, DNA extraction method were confirmed for the effective detection of M. leprae. And restriction enzyme fragment length polymorphism typing and high resolution melt (HRM) analysis were performed for comparison with sequencing analysis.
Methods: Compared with three DNA extraction methods (BB, SM and SP) with real-time polymerase chain reaction (PCR). Analysis single nucleotide polymorphism (SNP) genotype and tandem repeats by PCR amplification, and then compare with sequence.
Results: BB method was effective when measuring the concentration and threshold cycle (Ct) compared with SM and SP methods. When compared with restriction fragment length polymorphism typing method and sequence analysis, all methods were suitable for SNP1 and 3 type classification. Tandem repeats values of BB method were correspond to sequence analysis than SM and SP methods in HRM analysis.
Conclusions: The DNA extraction method by bead is useful approach for studying of M. leprae.
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KEYWORD
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DNA extraction, Leprosy, Mycobacterium leprae, PCR
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