Drawback of loop-mediated isothermal amplification
Deguo Wang, Guicheng Huo, Fugui Wang, Yonggang Li, Daxi Ren
Abstract
Deguo Wang, Guicheng Huo, Fugui Wang, Yonggang Li, Daxi Ren
Abstract
Loop-mediated isothermal amplification (LAMP) was a novel nucleic acid amplification method that amplified DNA with high specificity, efficiency and rapidity under isothermal conditions using a set of four specially designed primers and a DNA polymerase with strand displacement activity. In our research, LAMP was developed to detect foodborn Salmonella in raw milk, a set of six primers, two outer, two inner and two loop primers, were designed from Salmonella genomic DNA targeting invA. Temperature condition for detection of Salmonella was optimized to be 61°C. At the same time, LAMP was found to be effective when the template was pure; however, the usefulness of LAMP methods can be limited by the presence of inhibitors in the analysis of raw milk, unlike all the reports about LAMP, the sensitivity of LAMP was lower than that of PCR methods. Key words: Loop-mediated Isothermal Amplification, LAMP, foodborn Salmonella, raw milk, sensitivity.
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Loop-mediated isothermal amplification (LAMP) was a novel nucleic acid amplification method that amplified DNA with high specificity, efficiency and rapidity under isothermal conditions using a set of four specially designed primers and a DNA polymerase with strand displacement activity. In our research, LAMP was developed to detect foodborn Salmonella in raw milk, a set of six primers, two outer, two inner and two loop primers, were designed from Salmonella genomic DNA targeting invA. Temperature condition for detection of Salmonella was optimized to be 61°C. At the same time, LAMP was found to be effective when the template was pure; however, the usefulness of LAMP methods can be limited by the presence of inhibitors in the analysis of raw milk, unlike all the reports about LAMP, the sensitivity of LAMP was lower than that of PCR methods. Key words: Loop-mediated Isothermal Amplification, LAMP, foodborn Salmonella, raw milk, sensitivity.
Key concepts: Loop-mediated isothermal amplification, Multiple displacement amplification, Recombinase Polymerase Amplification, Salmonella, Isothermal process, Nucleic acid, DNA, Molecular biology