期刊
SENSORS AND ACTUATORS B-CHEMICAL
卷 369, 期 -, 页码 -出版社
ELSEVIER SCIENCE SA
DOI: 10.1016/j.snb.2022.132269
关键词
Isothermal amplification; High-efficiency transcription; T4 Polynucleotide Kinase phosphatase activity
资金
- National Natural Science Foundation of China [21705118]
- National Science Foundation of Tianjin [19ZXDBSY00070, 20YFZCSY00990]
In this study, a highly sensitive and selective transcription-based signal amplification method was developed for trace nucleic acid detection. The method utilizes the advantages of long template synthesis and self-amplification, and can be further modified for the detection of other analytical targets.
Signal amplification plays a significant role in nucleic acid analysis, especially in trace nucleic acid detection. Transcription-based signal amplification is a very important component in the field of signal amplification, and has received extensive attention depending on outstanding cyclic transcription ability. However, the low efficiency of transcription caused by limited template length usually restricts their further applications significantly. Herein, we developed a transcription-based signal amplification method with high sensitivity and selectivity. The approach we proposed couples the advantageous features of long template synthesis from phosphorothioated-terminal hairpin formation and self-priming extension (PS-THSP) with self-amplification from cell-free transcription system. The results demonstrate that this approach can be applied to quantify T4 Polynucleotide Kinase (T4 PNK) phosphatase activity in a broad range from 0.00001 to 0.01 U/mL, with a detection limit of 8.1 x 10(-6) U/mL. Moreover, this device can be further employed as a superior signal amplification platform for the detection of other analytical targets after moderate modifications.
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