Multiplex real-time PCR using temperature sensitive primer-supplying hydrogel particles and its application for malaria species identification

Title
Multiplex real-time PCR using temperature sensitive primer-supplying hydrogel particles and its application for malaria species identification
Authors
김상경김준선정승원변문섭유창훈심상준임채승김성우
Keywords
Real-time PCR; Multiplex; hydrogel particles; malaria; supplimer
Issue Date
2018-01
Publisher
PLoS ONE
Citation
VOL 13, NO 1-12
Abstract
Real-time PCR, also called quantitative PCR (qPCR), has been powerful analytical tool for detection of nucleic acids since it developed. Not only for biological research but also for diagnostic needs, qPCR technique requires capacity to detect multiple genes in recent years. Solid phase PCR (SP-PCR) where one or two directional primers are immobilized on solid substrates could analyze multiplex genetic targets. However, conventional SP-PCR was subjected to restriction of application for lack of PCR efficiency and quantitative resolution. Here we introduce an advanced qPCR with primer-incorporated network (PIN). One directional primers are immobilized in the porous hydrogel particle by covalent bond and the other direction of primers are temporarily immobilized at so-called 'Supplimers'. Supplimers released the primers to aqueous phase in the hydrogel at the thermal cycling of PCR. It induced the high PCR efficiency over 92% with high reliability. It reduced the formation of primer dimers and improved the selectivity of qPCR thanks to the strategy of 'right primers supplied to right place only'. By conducting a six-plex qPCR of 30 minutes, we analyzed DNA samples originated from malaria patients and successfully identified malaria species in a single reaction.
URI
http://pubs.kist.re.kr/handle/201004/67734
ISSN
1932-6203
Appears in Collections:
KIST Publication > Article
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