Multiplex real-time PCR using temperature sensitive primer-supplying hydrogel particles and its application for malaria species identification.
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)...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2018-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC5749795?pdf=render |
| _version_ | 1818296775993196544 |
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| author | Junsun Kim Seungwon Jung Mun Sub Byoun Changhoon Yoo Sang Jun Sim Chae Seung Lim Sung Woo Kim Sang Kyung Kim |
| author_facet | Junsun Kim Seungwon Jung Mun Sub Byoun Changhoon Yoo Sang Jun Sim Chae Seung Lim Sung Woo Kim Sang Kyung Kim |
| author_sort | Junsun Kim |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-13T04:08:54Z |
| format | Article |
| id | doaj.art-3eac99abf8d94755b89ee2ccc73fbed9 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-13T04:08:54Z |
| publishDate | 2018-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-3eac99abf8d94755b89ee2ccc73fbed92022-12-22T00:00:07ZengPublic Library of Science (PLoS)PLoS ONE1932-62032018-01-01131e019045110.1371/journal.pone.0190451Multiplex real-time PCR using temperature sensitive primer-supplying hydrogel particles and its application for malaria species identification.Junsun KimSeungwon JungMun Sub ByounChanghoon YooSang Jun SimChae Seung LimSung Woo KimSang Kyung KimReal-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.http://europepmc.org/articles/PMC5749795?pdf=render |
| spellingShingle | Junsun Kim Seungwon Jung Mun Sub Byoun Changhoon Yoo Sang Jun Sim Chae Seung Lim Sung Woo Kim Sang Kyung Kim Multiplex real-time PCR using temperature sensitive primer-supplying hydrogel particles and its application for malaria species identification. PLoS ONE |
| title | Multiplex real-time PCR using temperature sensitive primer-supplying hydrogel particles and its application for malaria species identification. |
| title_full | Multiplex real-time PCR using temperature sensitive primer-supplying hydrogel particles and its application for malaria species identification. |
| title_fullStr | Multiplex real-time PCR using temperature sensitive primer-supplying hydrogel particles and its application for malaria species identification. |
| title_full_unstemmed | Multiplex real-time PCR using temperature sensitive primer-supplying hydrogel particles and its application for malaria species identification. |
| title_short | Multiplex real-time PCR using temperature sensitive primer-supplying hydrogel particles and its application for malaria species identification. |
| title_sort | multiplex real time pcr using temperature sensitive primer supplying hydrogel particles and its application for malaria species identification |
| url | http://europepmc.org/articles/PMC5749795?pdf=render |
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