Ladder-shape melting temperature isothermal amplification of nucleic acids

Method summary The LMTIA technique for isothermal amplification of nucleic acids in vitro was developed in this study. The existing software for PCR design primers can be used for the design of LMTIA primers. The LMTIA assay was preliminarily tested to be more sensitive than the LAMP assay. A novel method, termed ladder-shape melting temperature isothermal amplification (LMTIA), was developed in this study. As a proof of concept, one pair of primers or two pairs of nested primers and a thermostable DNA polymerase were employed to amplify the internal transcribed spacer of Oryza sativa with the ladder-shape melting temperature curve. Our results demonstrated that the LMTIA assay with nested primers was 50-fold more sensitive than the LAMP assay with the same level of specificity. The LMTIA method has the potential to be used for the prevention and control of emerging epidemics caused by different types of pathogens.

Automated summary

A novel method called ladder-shape melting temperature isothermal amplification (LMTIA) was developed in this study, which was found to be more sensitive than the existing LAMP assay. This method has the potential to be used for prevention and control of emerging epidemics caused by different types of pathogens.