DNA Quantification via ICP-MS Using Lanthanide-Labeled Probes and Ligation-Mediated Amplification

The combination of lanthanide-tagged oligonucleotide probes with inductively coupled plasma mass spectrometry (ICP-MS) as the detection technique is a novel labeling and analysis strategy for heterogeneous nucleic acid quantification assays. We describe a hybridization assay based on biotin–streptav...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2014-01, Vol.86 (1), p.585-591
Main Authors: Brückner, Kathrin, Schwarz, Kathleen, Beck, Sebastian, Linscheid, Michael W
Format: Article
Language:English
Subjects:
Amplification
Analytical chemistry
Animals
Assaying
Biological
Cattle
Chain reactions
Chemical elements
Deoxyribonucleic acid
DNA
DNA - analysis
DNA - genetics
DNA Ligases - analysis
DNA Ligases - genetics
Elution
Enzymes
Hybridization
Lanthanoid Series Elements - chemistry
Mass spectrometry
Mass Spectrometry - methods
Nucleic Acid Hybridization - methods
Oligonucleotide Probes
Serum Albumin, Bovine - analysis
Serum Albumin, Bovine - genetics
Spectrophotometry, Atomic - methods
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Summary:The combination of lanthanide-tagged oligonucleotide probes with inductively coupled plasma mass spectrometry (ICP-MS) as the detection technique is a novel labeling and analysis strategy for heterogeneous nucleic acid quantification assays. We describe a hybridization assay based on biotin–streptavidin affinity using lanthanide-labeled reporter probes and biotinylated capture probes. For the basic sandwich type assay, performed in streptavidin-coated microtitration wells, the limit of detection (LOD) was 7.2 fmol of DNA target, corresponding to a final concentration of 6 pM terbium-labeled probes detectable by ICP-MS after elution from the solid support. To improve the sensitivity and sequence specificity of the approach, it was combined with established molecular biological techniques, i.e., elution with a restriction endonuclease and signal and target amplification by the ligase detection reaction (LDR) and ligase chain reaction (LCR), respectively. Initial experiments showed that the enzymes facilitated the discrimination of single-base mismatches within the recognition or ligation site. Furthermore, LCR as a target amplification step resulted in a 6000-fold increase of sensitivity, and finally an LOD of 2.6 amol was achieved with an artificial double-stranded DNA target.
ISSN:0003-2700
1520-6882
DOI:10.1021/ac402668p