Sequence-specific electrochemical detection of double-strand PCR amplicons of PML/RARα fusion gene in acute promyelocytic leukemia.
A novel electrochemical method for the sequence-specific detection of double-stranded polymerase chain reaction (PCR) products of PML/RARα fusion gene in acute promyelocytic leukemia (APL) was described in detail. Based on a "sandwich" sensing mode involving a pair of locked nucleic acids probes (capture probe and reporter probe), this DNA sensor exhibited excellent selectivity and specificity. The direct and quantitative analysis of double-stranded complementary was firstly performed by our sensor without the use of alkali, helicase enzymes, or denaturants. Finally, combining PCR technique with electrochemical detection scheme, PCR amplicons (191 bp) of the PML/RARα fusion gene were obtained and rapidly identified with a low detection limit of 79 fmol in the 100-μL hybridization system. The results clearly showed the power of sensor as a promising tool for the sensitive, specific, and portable detection of APL and other diseases.
Department of Pharmaceutical Analysis, Faculty of Pharmacy, Fujian Medical University, Fuzhou 350004, China.
Yun
Y
Lei
Mei-juan
MJ
Feng
Kun
K
Wang
Li-qing
LQ
Lin
Yuan-zhong
YZ
Chen
Xin-hua
XH
Lin
D015374:Biosensing Techniques; D019921:Biotinylation; D002138:Calibration; D004247:DNA / Q000737:chemistry; D004563:Electrochemistry / Q000379:methods*; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute / Q000378:metabolism*; D009363:Neoplasm Proteins / Q000737:chemistry; D009687:Nuclear Proteins / Q000378:metabolism*; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000235:genetics; D015514:Oncogene Proteins, Fusion / Q000737:chemistry; D016133:Polymerase Chain Reaction / Q000379:methods*; D000071156:Promyelocytic Leukemia Protein; D018168:Receptors, Retinoic Acid / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D000072482:Retinoic Acid Receptor alpha; D013997:Time Factors; D014157:Transcription Factors / Q000378:metabolism*; D025521:Tumor Suppressor Proteins / Q000378:metabolism*
D009363:Neoplasm Proteins; D009687:Nuclear Proteins; D009841:Oligonucleotides; D015514:Oncogene Proteins, Fusion; D000071156:Promyelocytic Leukemia Protein; C000607071:RARA protein, human; D018168:Receptors, Retinoic Acid; D011993:Recombinant Fusion Proteins; D000072482:Retinoic Acid Receptor alpha; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; C070576:PML protein, human; D004247:DNA