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ۼ : 16-05-16 11:36
[2016.05]ؿ Quantification of Fewer Than Ten Copies of a DNA Biomarker without Amplification or Labeling
 ۾ :
ȸ : 1,160  

Quantification of Fewer Than Ten Copies of a DNA Biomarker without Amplification or Labeling

 

Yoonhee Lee, Youngkyu Kim, Donggyu Lee, Dhruvajyoti Roy, and Joon Won Park



Polymerase chain reaction (PCR) is a highly sensitive diagnosis technique for detection of nucleic acids and for monitoring residual disease; however, PCR can be unreliable for samples containing very few target molecules. Here, we describe a quantification method, using force–distance (FD) curve based atomic force microscopy (AFM) to detect a target DNA bound to small (1.4–1.9 m diameter) probe DNA spots, allowing mapping of entire spots to nanometer resolution. Using a synthetic BCR-ABL fusion gene sequence target, we examined samples containing between one and 10 target copies. A high degree of correlation (r2 = 0.994) between numbers of target copies and detected probe clusters was observed, and the approach could detect the BCR-ABL biomarker when only a single copy was present, although multiple screens were required. Our results clearly demonstrate that FD curve-based imaging is suitable for quantitative analysis of fewer than 10 copies of DNA biomarkers without amplification, modification, or labeling.

 

̰ ȣۿ ̿Ͽ DNA biomarker ΰ ִ bioassay ÷ Ͽ. Probe DNA spot target DNA AFM Žħ DNA øӿ Ư ִ. ̷ biomarker BCR-ABL gene target Ͽ м 1 ǥ DNA ִ sample ̸ ȮϿ.

 

ش Ƿ о߿ ΰ ̴ ǽð ȿ (RT-qPCR) м ̸, Ư ̳ ǥ biomarker ִٴ ū ̴. ش biomarker мӸ ƴ϶ ū ޴ ع̷ biomarker ־ ϰ Ѵ. ̱ ȭȸ (Journal of the American Chemical Society) Ǿ.


 
 

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