Ladder-shape melting temperature isothermal amplification of nucleic acids
Deguo Wang,
Yongzhen Wang,
Meng Zhang,
Yongqing Zhang,
Juntao Sun,
Chunmei Song,
Fugang Xiao,
Yuan Ping,
Chen Pan,
Yushan Hu,
Chaoqun Wang,
Yanhong Liu
Affiliations
Deguo Wang
1Key Laboratory of Biomarker-Based Rapid Detection Technology for Food Safety of Henan Province, Xuchang University, Xuchang, 461000, China
Yongzhen Wang
1Key Laboratory of Biomarker-Based Rapid Detection Technology for Food Safety of Henan Province, Xuchang University, Xuchang, 461000, China
Meng Zhang
1Key Laboratory of Biomarker-Based Rapid Detection Technology for Food Safety of Henan Province, Xuchang University, Xuchang, 461000, China
Yongqing Zhang
1Key Laboratory of Biomarker-Based Rapid Detection Technology for Food Safety of Henan Province, Xuchang University, Xuchang, 461000, China
Juntao Sun
1Key Laboratory of Biomarker-Based Rapid Detection Technology for Food Safety of Henan Province, Xuchang University, Xuchang, 461000, China
Chunmei Song
1Key Laboratory of Biomarker-Based Rapid Detection Technology for Food Safety of Henan Province, Xuchang University, Xuchang, 461000, China
Fugang Xiao
1Key Laboratory of Biomarker-Based Rapid Detection Technology for Food Safety of Henan Province, Xuchang University, Xuchang, 461000, China
Yuan Ping
1Key Laboratory of Biomarker-Based Rapid Detection Technology for Food Safety of Henan Province, Xuchang University, Xuchang, 461000, China
Chen Pan
1Key Laboratory of Biomarker-Based Rapid Detection Technology for Food Safety of Henan Province, Xuchang University, Xuchang, 461000, China
Yushan Hu
1Key Laboratory of Biomarker-Based Rapid Detection Technology for Food Safety of Henan Province, Xuchang University, Xuchang, 461000, China
Chaoqun Wang
2Xuchang Customs, Xuchang, 461000, China
Yanhong Liu
3Molecular Characterization of Foodborne Pathogens Research Unit, Eastern Regional Research Center, Agricultural Research Service, US Department of Agriculture, Wyndmoor, PA 19038, USA
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.
