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

Junsun Kim, Seungwon Jung, Mun Sub Byoun, Changhoon Yoo, Sang Jun Sim, Chae Seung Lim, Sung Woo Kim, Sang Kyung Kim

Research output: Contribution to journalArticle

1 Citation (Scopus)

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.

Original languageEnglish
Article numbere0190451
JournalPLoS One
Volume13
Issue number1
DOIs
Publication statusPublished - 2018 Jan 1

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Hydrogel
Multiplex Polymerase Chain Reaction
hydrocolloids
malaria
Malaria
Real-Time Polymerase Chain Reaction
quantitative polymerase chain reaction
Polymerase Chain Reaction
Temperature
temperature
Covalent bonds
Thermal cycling
Dimers
Nucleic Acids
Genes
DNA
Substrates
nucleic acids
species identification
heat

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Multiplex real-time PCR using temperature sensitive primer-supplying hydrogel particles and its application for malaria species identification. / Kim, Junsun; Jung, Seungwon; Byoun, Mun Sub; Yoo, Changhoon; Sim, Sang Jun; Lim, Chae Seung; Kim, Sung Woo; Kim, Sang Kyung.

In: PLoS One, Vol. 13, No. 1, e0190451, 01.01.2018.

Research output: Contribution to journalArticle

Kim, Junsun ; Jung, Seungwon ; Byoun, Mun Sub ; Yoo, Changhoon ; Sim, Sang Jun ; Lim, Chae Seung ; Kim, Sung Woo ; Kim, Sang Kyung. / Multiplex real-time PCR using temperature sensitive primer-supplying hydrogel particles and its application for malaria species identification. In: PLoS One. 2018 ; Vol. 13, No. 1.
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