Tags

Type your tag names separated by a space and hit enter

Real-time ligation chain reaction for DNA quantification and identification on the FO-SPR.
Biosens Bioelectron. 2015 May 15; 67:394-9.BB

Abstract

Different assays have been developed in the past years to meet point-of-care diagnostic tests requirements for fast and sensitive quantification and identification of targets. In this paper, we developed the ligation chain reaction (LCR) assay on the Fiber Optic Surface Plasmon Resonance (FO-SPR) platform, which enabled simultaneous quantification and cycle-to-cycle identification of DNA during amplification. The newly developed assay incorporated FO-SPR DNA melting assay, previously developed by our group. This required establishment of several assay parameters, including buffer ionic strength and thermal ramping speed as these parameters both influence the ligation enzyme performance and the hybridization yield of the gold nanoparticles (Au NPs) on the FO-SPR sensor. Quantification and identification of DNA targets was achieved over a wide concentration range with a calibration curve spanning 7 orders of magnitude and LOD of 13.75 fM. Moreover, the FO-SPR LCR assay could discriminate single nucleotide polymorphism (SNPs) without any post reaction analysis, featuring thus all the essential requirements of POC tests.

Links

Publisher Full Text (DOI)

Authors+Show Affiliations

Knez K
BIOSYST-MeBioS, KU Leuven - University of Leuven, Willem de Croylaan 42, B-3001 Leuven, Belgium. Electronic address: knez.karel@gmail.com.
Spasic D
BIOSYST-MeBioS, KU Leuven - University of Leuven, Willem de Croylaan 42, B-3001 Leuven, Belgium. Electronic address: Dragana.Spasic@biw.kuleuven.be.
Delport F
BIOSYST-MeBioS, KU Leuven - University of Leuven, Willem de Croylaan 42, B-3001 Leuven, Belgium. Electronic address: Filip.Delport@biw.kuleuven.be.
Lammertyn J
BIOSYST-MeBioS, KU Leuven - University of Leuven, Willem de Croylaan 42, B-3001 Leuven, Belgium. Electronic address: jeroen.lammertyn@biw.kuleuven.be.

MeSH

Biosensing TechniquesDNAFiber Optic TechnologyGoldHumansLigase Chain ReactionMetal NanoparticlesNucleic Acid Amplification TechniquesNucleic Acid DenaturationPolymorphism, Single NucleotideSurface Plasmon Resonance

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

25212376

Citation

Knez, Karel, et al. "Real-time Ligation Chain Reaction for DNA Quantification and Identification On the FO-SPR." Biosensors & Bioelectronics, vol. 67, 2015, pp. 394-9.
Knez K, Spasic D, Delport F, et al. Real-time ligation chain reaction for DNA quantification and identification on the FO-SPR. Biosens Bioelectron. 2015;67:394-9.
Knez, K., Spasic, D., Delport, F., & Lammertyn, J. (2015). Real-time ligation chain reaction for DNA quantification and identification on the FO-SPR. Biosensors & Bioelectronics, 67, 394-9. https://doi.org/10.1016/j.bios.2014.08.067
Knez K, et al. Real-time Ligation Chain Reaction for DNA Quantification and Identification On the FO-SPR. Biosens Bioelectron. 2015 May 15;67:394-9. PubMed PMID: 25212376.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Real-time ligation chain reaction for DNA quantification and identification on the FO-SPR. AU - Knez,Karel, AU - Spasic,Dragana, AU - Delport,Filip, AU - Lammertyn,Jeroen, Y1 - 2014/09/01/ PY - 2014/06/06/received PY - 2014/08/26/revised PY - 2014/08/27/accepted PY - 2014/9/13/entrez PY - 2014/9/13/pubmed PY - 2015/10/27/medline KW - FO-SPR melting assay KW - Fiber Optic Surface Plasmon Resonance (FO-SPR) KW - Gold nanoparticles (Au NPs) KW - Ligation chain reaction (LCR) SP - 394 EP - 9 JF - Biosensors & bioelectronics JO - Biosens Bioelectron VL - 67 N2 - Different assays have been developed in the past years to meet point-of-care diagnostic tests requirements for fast and sensitive quantification and identification of targets. In this paper, we developed the ligation chain reaction (LCR) assay on the Fiber Optic Surface Plasmon Resonance (FO-SPR) platform, which enabled simultaneous quantification and cycle-to-cycle identification of DNA during amplification. The newly developed assay incorporated FO-SPR DNA melting assay, previously developed by our group. This required establishment of several assay parameters, including buffer ionic strength and thermal ramping speed as these parameters both influence the ligation enzyme performance and the hybridization yield of the gold nanoparticles (Au NPs) on the FO-SPR sensor. Quantification and identification of DNA targets was achieved over a wide concentration range with a calibration curve spanning 7 orders of magnitude and LOD of 13.75 fM. Moreover, the FO-SPR LCR assay could discriminate single nucleotide polymorphism (SNPs) without any post reaction analysis, featuring thus all the essential requirements of POC tests. SN - 1873-4235 UR - https://www.unboundmedicine.com/medline/citation/25212376/Real_time_ligation_chain_reaction_for_DNA_quantification_and_identification_on_the_FO_SPR_ DB - PRIME DP - Unbound Medicine ER -