PMID- 25356523
OWN - NLM
STAT- MEDLINE
DCOM- 20150908
LR  - 20141118
IS  - 1520-6882 (Electronic)
IS  - 0003-2700 (Linking)
VI  - 86
IP  - 22
DP  - 2014 Nov 18
TI  - Quencher-free fluorescent method for homogeneously sensitive detection of 
      microRNAs in human lung tissues.
PG  - 11410-6
LID - 10.1021/ac503365z [doi]
AB  - Sensitive and accurate analysis of microRNA (miRNA) expression is imperative for 
      understanding the biological functions of miRNAs and the early diagnosis of human 
      cancers. Here, we develop a quencher-free fluorescent method for homogeneously 
      sensitive detection of let-7a miRNA using the target-triggered recycling signal 
      amplification in combination with a 2-aminopurine probe. The 2-aminopurine probe 
      is characterized by the substitution of 2-aminopurine for adenine in the DNA 
      strand and the quenching of 2-aminopurine fluorescence through its stacking 
      interaction with the adjacent bases. The binding of target miRNA with the 
      2-aminopurine probe initiates the extension reaction in the presence of 
      polymerase to produce the DNA duplexes. These DNA duplexes can be further cleaved 
      by lambda exonuclease through the recycling digestion to release abundant free 
      2-aminopurines, leading to an enhanced fluorescence signal. The proposed method 
      exhibits high sensitivity with a detection limit of 0.3 fmol, and it can even 
      discriminate the single-base difference among the miRNA family members. More 
      importantly, this method can accurately distinguish the expression of let-7a 
      miRNA in human lung tissues between ten non small cell lung cancer (NSCLC) 
      patients and ten healthy persons, holding a great potential for further 
      application in early clinical diagnosis.
FAU - Zhu, Guichi
AU  - Zhu G
AD  - Single-Molecule Detection and Imaging Laboratory, Shenzhen Institutes of Advanced 
      Technology, Chinese Academy of Sciences , Shenzhen, Guangdong 518055, China.
FAU - Liang, Li
AU  - Liang L
FAU - Zhang, Chun-yang
AU  - Zhang CY
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20141107
PL  - United States
TA  - Anal Chem
JT  - Analytical chemistry
JID - 0370536
RN  - 0 (Fluorescent Dyes)
RN  - 0 (MicroRNAs)
RN  - 452-06-2 (2-Aminopurine)
SB  - IM
MH  - 2-Aminopurine/chemistry
MH  - *Fluorescence
MH  - Fluorescent Dyes/chemistry
MH  - Humans
MH  - Lung/*metabolism
MH  - MicroRNAs/*analysis
EDAT- 2014/10/31 06:00
MHDA- 2015/09/09 06:00
CRDT- 2014/10/31 06:00
PHST- 2014/10/31 06:00 [entrez]
PHST- 2014/10/31 06:00 [pubmed]
PHST- 2015/09/09 06:00 [medline]
AID - 10.1021/ac503365z [doi]
PST - ppublish
SO  - Anal Chem. 2014 Nov 18;86(22):11410-6. doi: 10.1021/ac503365z. Epub 2014 Nov 7.