Open AccessLadder-shape melting temperature isothermal amplification of nucleic acids
Author(s) -
Deguo Wang,
Yongzhen Wang,
Meng Zhang,
Yongqing Zhang,
Jian Sun,
Chao Song,
Fugang Xiao,
Yuan Ping,
Chen Pan,
Yushan Hu,
Chaoqun Wang,
Yanhong Liu
Publication year - 2021
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/btn-2020-0173
Subject(s) - loop mediated isothermal amplification , melting temperature , nucleic acid , melting curve analysis , isothermal process , polymerase chain reaction , nested polymerase chain reaction , dna , biology , microbiology and biotechnology , polymerase , genetics , chemistry , materials science , thermodynamics , physics , gene , composite material
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.