Isothermal Detection of DNA by Beacon-Assisted Detection Amplification
Isothermal Detection of DNA by Beacon-Assisted Detection Amplification
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DOI：
10.1002/anie.200906992
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发表时间：
2010-01-01
影响因子：
16.6
通讯作者：
Trau, Matt
中科院分区：
文献类型：
--
作者：
Connolly, Ashley R.;Trau, Matt
Nucleic acids are routinely used as biomarkers to help diagnose pathogenic infections and genetic disorders. Specific nucleic acids indicating the presence of a disease are often found in only trace amounts in a complex biological extract, so new technologies to detect a unique DNA sequence amongst the millions of nucleotides that constitute a genome are constantly in demand. Analysis of DNA is currently performed by amplifying trace amounts of a specific sequence to levels that are detectable. Several DNA amplification techniques have been developed which can be broadly divided into those that require thermal cycling and those that proceed at constant temperature (isothermal). The polymerase chain reaction (PCR) is the most widely used thermal cycling technique for DNA amplification.[1] The reaction proceeds exponentially so trace amounts of DNA can be amplified to detectable levels. However, thermal cycling imposes instrumental constraints that limit this technique to a laboratory setting, and the PCR product typically requires characterization at the end of the reaction to determine the specificity of amplification. Isothermal amplification of DNA has emerged as an alternative amplification technique that often employs a strand displacement polymerase to continuously replicate one strand of a DNA duplex.[2–8] The reaction proceeds at constant temperature, so the time required for DNA amplification is less than that required for thermal cycling techniques. Reactions can be performed without specialized instrumentation and have the potential for “on the spot” testing of DNA. However, the isothermal nature of these reactions can result in non-specific priming and the production of unwanted byproducts.[9] While strategies to make isothermal amplification more specific have been developed,[10–14] advances in the accuracy, sensitivity, speed and simplicity of DNA detection would be beneficial in the laboratory and for “on the spot” testing of DNA.
