Homogeneous and sensitive detection of microRNA with ligase chain reaction and lambda exonuclease-assisted cationic conjugated polymer biosensing.
A simple and homogeneous microRNA assay is developed by integration of ligase chain reaction (LCR) and lambda exonuclease-assisted cationic conjugated polymer (CCP) biosensing. LCR is utilized for exponential amplification of microRNA, and lambda exonuclease is introduced to degrade excess fluorescein-labeled probes in LCR for eliminating background signal. After addition of CCP, efficient fluorescence resonance energy transfer from CCP to fluorescein in LCR products occurs. The method is sensitive enough to detect 0.1 fM target microRNA and specific to discriminate one-base difference of microRNAs, which paves a new way for homogeneous microRNA detection and molecular diagnosis.
Key Laboratory of Medicinal Chemistry and Molecular Diagnosis, Ministry of Education, College of Chemistry and Environmental Science, Hebei University , Baoding 071002, China.
Zheng
Z
Yuan
Yuanyuan
Y
Zhou
Shuxin
S
Gao
Yongqiang
Y
Cheng
Zhengping
Z
Li
D015374:Biosensing Techniques; D002412:Cations; D005092:Exonucleases / Q000737:chemistry*; D019793:Fluorescein / Q000737:chemistry; D031541:Fluorescence Resonance Energy Transfer; D022521:Ligase Chain Reaction; D057230:Limit of Detection; D035683:MicroRNAs / Q000032:analysis*; D011108:Polymers / Q000032:analysis*; D013050:Spectrometry, Fluorescence
D002412:Cations; D035683:MicroRNAs; D011108:Polymers; D005092:Exonucleases; D019793:Fluorescein