Microparticle-based RT-qPCR for highly selective rare mutation detection

Title
Microparticle-based RT-qPCR for highly selective rare mutation detection
Authors
김상경오은해정승원김원진김광표
Keywords
RT-qPCR; rimer-immobilized particles; Bcr-Abl fusion transcript
Issue Date
2017-01
Publisher
Biosensors and bioelectronics
Citation
VOL 87, NO 1-235
Abstract
The quantitative reverse transcription polymerase chain reaction (RT-qPCR) has become one of the most widely used methods in the detection of disease-specific RNAs. The RT-qPCR involves two separate steps, RT and qPCR. In this study, we suggest a new RT-qPCR protocol with the particles of primer-immobilized networks (PINs), performing capture, RT and amplification of a target RNA in one particle. The production of undesired cDNAs was dramatically suppressed by the specific capture of the target RNA within the particle. Afterward, RT and amplification processes are performed without loss of cDNAs as exchanging the reaction solution. The biomarker gene of chronic myeloid leukemia, Bcr-Abl fusion transcript, is detected in the sensitivity of single mutant leukemic cell mixed in 104 normal cell using this protocol with the excellent restraint of non-specific signal. This protocol that whole processes are performed in the particle in a row is preferred for the highly specific detection of target RNAs in complex sample.
URI
http://pubs.kist.re.kr/handle/201004/64913
ISSN
0956-5663
Appears in Collections:
KIST Publication > Article
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