SENSITIVE DETECTION OF NUCLEIC ACID BY SIGNAL AMPLIFICATION APPROACH BASED ON POLYMERIZATION COUPLED WITH ATP AMPLIFICATION

摘要：

Detection of nucleic acid has many benefits in molecular diagnosis. To detect low concentration single strand DNA (ssDNA), a novel signal amplification approach based on DNA polymerization and ATP amplification reaction was developed. In this approach, target ssDNA annealing with a primer was extended by polymerase to release inorganic pyrophosphate (PPi), which was subsequently converted to ATP by ATP sulfurylase with its substrate adenosine phosphosulfate (APS). The resulting ATP was amplified by a fusion protein adenylate kinase and polyphosphate kinase (ADK-PPK) catalyzed ATP amplification reaction, and measured through bioluminescence assay. After decreasing the background signal with enzymatic method, as low as 5. 2 frnol/L ssDNA was successfully detected. However, some efforts need to do to achieve reproducibility and higher sensitivity. This is a dye-free and low cost signal amplification approach to detect ssDNA.

关键词： Signal amplification ATP amplification DNA polymerization

作者: Zou Bingjie Chen Ying Huang Huan Zhou Guohua

作者单位: Huadong Research Institute for Medicine and Biotechnics, Nanjing 210002, China School of Life Scienc Huadong Research Institute for Medicine and Biotechnics, Nanjing 210002, China Huadong Research Institute for Medicine and Biotechnics, Nanjing 210002, China School of Life Scienc

会议类型: 国际会议

会议名称: The 6th International Forum on Post-genome Technologies(6IFPT)(第六届国际后基因组生命科学技术学术论坛)

会议地点: 北京

会议语种:英文

页码: 431-433

在线出版日期: 2009-09-17（万方平台首次上网日期，不代表论文的发表时间）