Addition of dNTPs can improve the detection sensitivity of catalytic hairpin assembly
Mingyuan Zou,
Meiling Zhou,
Shuo Ma,
Chen Zhang,
Feng Xiao,
Huina Wu,
Abudushalamu GuliNaizhaer,
Yuming Yao,
Yaya Chen,
Shijie Cai,
Xiaobo Fan,
Fengfeng Zhao,
Guoqiu Wu
Affiliations
Mingyuan Zou
Southeast University, Zhongda Hospital, Center of Clinical Laboratory Medicine, Medical School, Nanjing 210009, People’s Republic of China
Meiling Zhou
Southeast University, Zhongda Hospital, Center of Clinical Laboratory Medicine, Medical School, Nanjing 210009, People’s Republic of China
Shuo Ma
Southeast University, Zhongda Hospital, Center of Clinical Laboratory Medicine, Medical School, Nanjing 210009, People’s Republic of China
Chen Zhang
Southeast University, Zhongda Hospital, Center of Clinical Laboratory Medicine, Medical School, Nanjing 210009, People’s Republic of China; Public Health School, Southeast University, Nanjing 210009, People’s Republic of China
Feng Xiao
Southeast University, Zhongda Hospital, Center of Clinical Laboratory Medicine, Medical School, Nanjing 210009, People’s Republic of China
Huina Wu
Southeast University, Zhongda Hospital, Center of Clinical Laboratory Medicine, Medical School, Nanjing 210009, People’s Republic of China
Abudushalamu GuliNaizhaer
Southeast University, Zhongda Hospital, Center of Clinical Laboratory Medicine, Medical School, Nanjing 210009, People’s Republic of China
Yuming Yao
Southeast University, Zhongda Hospital, Center of Clinical Laboratory Medicine, Medical School, Nanjing 210009, People’s Republic of China
Yaya Chen
Southeast University, Zhongda Hospital, Center of Clinical Laboratory Medicine, Medical School, Nanjing 210009, People’s Republic of China
Shijie Cai
Southeast University, Zhongda Hospital, Center of Clinical Laboratory Medicine, Medical School, Nanjing 210009, People’s Republic of China
Xiaobo Fan
Department of Clinical Laboratory Medicine, Medical School, Southeast University, Nanjing 210009, People’s Republic of China
Fengfeng Zhao
Southeast University, Zhongda Hospital, Center of Clinical Laboratory Medicine, Medical School, Nanjing 210009, People’s Republic of China; Department of Clinical Laboratory Medicine, Medical School, Southeast University, Nanjing 210009, People’s Republic of China; Corresponding author
Guoqiu Wu
Southeast University, Zhongda Hospital, Center of Clinical Laboratory Medicine, Medical School, Nanjing 210009, People’s Republic of China; Department of Clinical Laboratory Medicine, Medical School, Southeast University, Nanjing 210009, People’s Republic of China; Jiangsu Provincial Key Laboratory of Critical Care Medicine, Department of Diagnostics, School of Medicine, Southeast University, Nanjing, Jiangsu 210009, China; Corresponding author
Summary: Ever since the catalytic hairpin assembly (CHA) circuit has been highlighted as a powerful nucleic acid detection tool, nucleic acid detection methods based on CHA have been widely studied. However, the detection sensitivity of CHA-based methods is insufficient. The relatively high background signals resulting from the spontaneous reaction between the two hairpin probes is one of the major reasons for limiting the sensitivity of CHA. In this study, we established that the addition of deoxynucleotide triphosphates (dNTPs) to the reaction system can significantly reduce the background leakage of CHA. The dNTPs-CHA, coupled with a fluorescence lateral flow assay strip, is used for the rapid and highly sensitive detection of miRNA. It is capable of reliably detecting miRNA in serum samples down to a limit of 100 aM, which is an improvement in the lower detection limit by nearly five orders of magnitude compared to that of the pure CHA.
