Enhancing melting curve analysis for the discrimination of loop-mediated isothermal amplification products from four pathogenic molds: Use of inorganic pyrophosphatase and its effect in reducing the variance in melting temperature values

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• 作者：Kazuya Tone^a ; ^b ; Ryuichi Fujisaki^c ; Takashi Yamazaki^a ; ^b ; ^d ; Koichi Makimura^a ; ^b ; ^d ; ^{makimura@med.teikyo-u.ac.jp}
• 关键词：LAMP ; loop-mediated isothermal amplification ; PCR ; polymerase chain reaction ; MCA ; melting curve analysis ; PPase ; thermostable inorganic pyrophosphatase ; Tm ; melting temperature ; Afum ; Aspergillus fumigatus ; Pexp ; Penicillium expansum ; Pmar ; Penicillium marneffei ; Hcap ; Histoplasma capsulatum
• 刊名：Journal of Microbiological Methods
• 出版年：2017
• 出版时间：January 2017
• 年：2017
• 卷：132
• 期：Complete
• 页码：41-45
• 全文大小：1355 K
• 卷排序：132

文摘

LAMP is widely used for differentiating causative agents in infectious diseases. MCA with LAMP reduces both the labor required to conduct an assay and contamination. We investigated the influence of PPase on the melting temperature of LAMP products. Use of PPase reduced melting temperature variance and helped differentiate pathogens. Tm values of LAMP products increase in accordance with the GC contents of the starting structure.