TY - JOUR
T1 - Quantification of Fewer than Ten Copies of a DNA Biomarker without Amplification or Labeling
AU - Lee, Yoonhee
AU - Kim, Youngkyu
AU - Lee, Donggyu
AU - Roy, Dhruvajyoti
AU - Park, Joon Won
N1 - Publisher Copyright:
© 2016 American Chemical Society.
PY - 2016/6/8
Y1 - 2016/6/8
N2 - 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 μm 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.
AB - 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 μm 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.
UR - http://www.scopus.com/inward/record.url?scp=84974791430&partnerID=8YFLogxK
U2 - 10.1021/jacs.6b02791
DO - 10.1021/jacs.6b02791
M3 - Article
C2 - 27175474
AN - SCOPUS:84974791430
SN - 0002-7863
VL - 138
SP - 7075
EP - 7081
JO - Journal of the American Chemical Society
JF - Journal of the American Chemical Society
IS - 22
ER -