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A dumbbell probe-mediated rolling circle amplification strategy for highly sensitive microRNA detection.

Zhou Y, Huang Q, Gao J, Lu J, Shen X, Fan C - Nucleic Acids Res. (2010)

Bottom Line: The femtomolar sensitivity of D-RCA compares favorably with other existing technologies.More significantly, the dynamic range of D-RCA is extremely large, covering eight orders of magnitude.We also demonstrate miRNA quantification with this highly sensitive and inexpensive D-RCA strategy in clinical samples.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Physical Biology, Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai, China.

ABSTRACT
We herein report the design of a dumbbell-shaped DNA probe that integrates target-binding, amplification and signaling within one multifunctional design. The dumbbell probe can initiate rolling circle amplification (D-RCA) in the presence of specific microRNA (miRNA) targets. This D-RCA-based miRNA strategy allows quantification of miRNA with very low quantity of RNA samples. The femtomolar sensitivity of D-RCA compares favorably with other existing technologies. More significantly, the dynamic range of D-RCA is extremely large, covering eight orders of magnitude. We also demonstrate miRNA quantification with this highly sensitive and inexpensive D-RCA strategy in clinical samples.

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Comparison of the detection performance by using either linear or dumbbell probes (12- and 24-base stems).
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Figure 2: Comparison of the detection performance by using either linear or dumbbell probes (12- and 24-base stems).

Mentions: Subsequently, we compared dumbbell probe-mediated RCA with linear-probe-mediated RCA to reveal the advantages of the D-RCA strategy (Figure 2 and Supplementary Figure S3). As observed for linear probe-mediated RCA, the signal was very low until the target concentration reached 1 pM. In contrast, the signal of dumbbell probe-mediated RCA was significantly higher than negative control when the target concentration was only 1 fM, exhibiting a sensitivity improvement by three orders of magnitude.Figure 2.


A dumbbell probe-mediated rolling circle amplification strategy for highly sensitive microRNA detection.

Zhou Y, Huang Q, Gao J, Lu J, Shen X, Fan C - Nucleic Acids Res. (2010)

Comparison of the detection performance by using either linear or dumbbell probes (12- and 24-base stems).
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2926626&req=5

Figure 2: Comparison of the detection performance by using either linear or dumbbell probes (12- and 24-base stems).
Mentions: Subsequently, we compared dumbbell probe-mediated RCA with linear-probe-mediated RCA to reveal the advantages of the D-RCA strategy (Figure 2 and Supplementary Figure S3). As observed for linear probe-mediated RCA, the signal was very low until the target concentration reached 1 pM. In contrast, the signal of dumbbell probe-mediated RCA was significantly higher than negative control when the target concentration was only 1 fM, exhibiting a sensitivity improvement by three orders of magnitude.Figure 2.

Bottom Line: The femtomolar sensitivity of D-RCA compares favorably with other existing technologies.More significantly, the dynamic range of D-RCA is extremely large, covering eight orders of magnitude.We also demonstrate miRNA quantification with this highly sensitive and inexpensive D-RCA strategy in clinical samples.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Physical Biology, Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai, China.

ABSTRACT
We herein report the design of a dumbbell-shaped DNA probe that integrates target-binding, amplification and signaling within one multifunctional design. The dumbbell probe can initiate rolling circle amplification (D-RCA) in the presence of specific microRNA (miRNA) targets. This D-RCA-based miRNA strategy allows quantification of miRNA with very low quantity of RNA samples. The femtomolar sensitivity of D-RCA compares favorably with other existing technologies. More significantly, the dynamic range of D-RCA is extremely large, covering eight orders of magnitude. We also demonstrate miRNA quantification with this highly sensitive and inexpensive D-RCA strategy in clinical samples.

Show MeSH