Real time quantitative amplification detection on a microarray : towards high multiplex quantitative PCR

Anke Pierik, M. Boamfa, M. Zelst, van, D. Clout, H.R. Stapert, J.F. Dijksman, D.J. Broer, R. Wimberger-Friedl

Research output: Contribution to journalArticleAcademicpeer-review

7 Citations (Scopus)
3 Downloads (Pure)

Abstract

Quantitative real-time polymerase chain reaction (qrtPCR) is widely used as a research and diagnostic tool. Notwithstanding its many powerful features, the method is limited in the degree of multiplexing to about 6 due to spectral overlap of the available fluorophores. A new method is presented that allows quantitative amplification detection at higher multiplexing by the integration of amplification in solution and monitoring via hybridization to a microarray in real-time. This method does not require any manipulation of the PCR product and runs in a single closed chamber. Employing labeled primers, one of the main challenges is to measure surface signals against a high fluorescence background from solution. A compact, confocal scanner is employed, based on miniaturized optics from DVD technology and combined with a flat thermocycler for simultaneous scanning and heating. The feasibility of this method is demonstrated in singleplex with an analytical sensitivity comparable to routine qrtPCR.
Original languageEnglish
Pages (from-to)18997-1902
Number of pages6
JournalLab on a Chip
Volume12
Issue number10
DOIs
Publication statusPublished - 2012

Fingerprint

Multiplex Polymerase Chain Reaction
Polymerase chain reaction
Microarrays
Multiplexing
Amplification
Videodisks
Fluorophores
Real-Time Polymerase Chain Reaction
Optics
Fluorescence
Scanning
Heating
Monitoring
Technology
Polymerase Chain Reaction
Research

Cite this

Pierik, A., Boamfa, M., Zelst, van, M., Clout, D., Stapert, H. R., Dijksman, J. F., ... Wimberger-Friedl, R. (2012). Real time quantitative amplification detection on a microarray : towards high multiplex quantitative PCR. Lab on a Chip, 12(10), 18997-1902. https://doi.org/10.1039/c2lc20740k
Pierik, Anke ; Boamfa, M. ; Zelst, van, M. ; Clout, D. ; Stapert, H.R. ; Dijksman, J.F. ; Broer, D.J. ; Wimberger-Friedl, R. / Real time quantitative amplification detection on a microarray : towards high multiplex quantitative PCR. In: Lab on a Chip. 2012 ; Vol. 12, No. 10. pp. 18997-1902.
@article{d694b570887b4e82bc809cf2118ec1b5,
title = "Real time quantitative amplification detection on a microarray : towards high multiplex quantitative PCR",
abstract = "Quantitative real-time polymerase chain reaction (qrtPCR) is widely used as a research and diagnostic tool. Notwithstanding its many powerful features, the method is limited in the degree of multiplexing to about 6 due to spectral overlap of the available fluorophores. A new method is presented that allows quantitative amplification detection at higher multiplexing by the integration of amplification in solution and monitoring via hybridization to a microarray in real-time. This method does not require any manipulation of the PCR product and runs in a single closed chamber. Employing labeled primers, one of the main challenges is to measure surface signals against a high fluorescence background from solution. A compact, confocal scanner is employed, based on miniaturized optics from DVD technology and combined with a flat thermocycler for simultaneous scanning and heating. The feasibility of this method is demonstrated in singleplex with an analytical sensitivity comparable to routine qrtPCR.",
author = "Anke Pierik and M. Boamfa and {Zelst, van}, M. and D. Clout and H.R. Stapert and J.F. Dijksman and D.J. Broer and R. Wimberger-Friedl",
year = "2012",
doi = "10.1039/c2lc20740k",
language = "English",
volume = "12",
pages = "18997--1902",
journal = "Lab on a Chip",
issn = "1473-0197",
publisher = "Royal Society of Chemistry",
number = "10",

}

Pierik, A, Boamfa, M, Zelst, van, M, Clout, D, Stapert, HR, Dijksman, JF, Broer, DJ & Wimberger-Friedl, R 2012, 'Real time quantitative amplification detection on a microarray : towards high multiplex quantitative PCR', Lab on a Chip, vol. 12, no. 10, pp. 18997-1902. https://doi.org/10.1039/c2lc20740k

Real time quantitative amplification detection on a microarray : towards high multiplex quantitative PCR. / Pierik, Anke; Boamfa, M.; Zelst, van, M.; Clout, D.; Stapert, H.R.; Dijksman, J.F.; Broer, D.J.; Wimberger-Friedl, R.

In: Lab on a Chip, Vol. 12, No. 10, 2012, p. 18997-1902.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Real time quantitative amplification detection on a microarray : towards high multiplex quantitative PCR

AU - Pierik, Anke

AU - Boamfa, M.

AU - Zelst, van, M.

AU - Clout, D.

AU - Stapert, H.R.

AU - Dijksman, J.F.

AU - Broer, D.J.

AU - Wimberger-Friedl, R.

PY - 2012

Y1 - 2012

N2 - Quantitative real-time polymerase chain reaction (qrtPCR) is widely used as a research and diagnostic tool. Notwithstanding its many powerful features, the method is limited in the degree of multiplexing to about 6 due to spectral overlap of the available fluorophores. A new method is presented that allows quantitative amplification detection at higher multiplexing by the integration of amplification in solution and monitoring via hybridization to a microarray in real-time. This method does not require any manipulation of the PCR product and runs in a single closed chamber. Employing labeled primers, one of the main challenges is to measure surface signals against a high fluorescence background from solution. A compact, confocal scanner is employed, based on miniaturized optics from DVD technology and combined with a flat thermocycler for simultaneous scanning and heating. The feasibility of this method is demonstrated in singleplex with an analytical sensitivity comparable to routine qrtPCR.

AB - Quantitative real-time polymerase chain reaction (qrtPCR) is widely used as a research and diagnostic tool. Notwithstanding its many powerful features, the method is limited in the degree of multiplexing to about 6 due to spectral overlap of the available fluorophores. A new method is presented that allows quantitative amplification detection at higher multiplexing by the integration of amplification in solution and monitoring via hybridization to a microarray in real-time. This method does not require any manipulation of the PCR product and runs in a single closed chamber. Employing labeled primers, one of the main challenges is to measure surface signals against a high fluorescence background from solution. A compact, confocal scanner is employed, based on miniaturized optics from DVD technology and combined with a flat thermocycler for simultaneous scanning and heating. The feasibility of this method is demonstrated in singleplex with an analytical sensitivity comparable to routine qrtPCR.

U2 - 10.1039/c2lc20740k

DO - 10.1039/c2lc20740k

M3 - Article

C2 - 22473033

VL - 12

SP - 18997

EP - 11902

JO - Lab on a Chip

JF - Lab on a Chip

SN - 1473-0197

IS - 10

ER -

Pierik A, Boamfa M, Zelst, van M, Clout D, Stapert HR, Dijksman JF et al. Real time quantitative amplification detection on a microarray : towards high multiplex quantitative PCR. Lab on a Chip. 2012;12(10):18997-1902. https://doi.org/10.1039/c2lc20740k