title: Quantification of DNA using the luminescent oxygen channeling assay.
abstract: BACKGROUND
  Simplified and cost-effective methods for the detection and quantification of nucleic acid targets are still a challenge in molecular diagnostics.
  
  
  METHODS
  Luminescent oxygen channeling assay (LOCI(TM)) latex particles can be conjugated to synthetic oligodeoxynucleotides and hybridized, via linking probes, to different DNA targets. These oligomer-conjugated LOCI particles survive thermocycling in a PCR reaction and allow quantified detection of DNA targets in both real-time and endpoint formats. The endpoint DNA quantification format utilized two sensitizer bead types that are sensitive to separate illumination wavelengths. These two bead types were uniquely annealed to target or control amplicons, and separate illuminations generated time-resolved chemiluminescence, which distinguished the two amplicon types.
  
  
  RESULTS
  In the endpoint method, ratios of the two signals allowed determination of the target DNA concentration over a three-log range. The real-time format allowed quantification of the DNA target over a six-log range with a linear relationship between threshold cycle and log of the number of DNA targets.
  
  
  CONCLUSIONS
  This is the first report of the use of an oligomer-labeled latex particle assay capable of producing DNA quantification and sequence-specific chemiluminescent signals in a homogeneous format. It is also the first report of the generation of two signals from a LOCI assay. The methods described here have been shown to be easily adaptable to new DNA targets because of the generic nature of the oligomer-labeled LOCI particles.
authors.affiliation: Advanced Diagnostics Group, Dade Behring Inc., PO Box 49013, San Jose, CA 95161-9013, USA.
authors.forename: R
authors.initials: R
authors.lastname: Patel
authors.forename: R
authors.initials: R
authors.lastname: Pollner
authors.forename: S
authors.initials: S
authors.lastname: de Keczer
authors.forename: J
authors.initials: J
authors.lastname: Pease
authors.forename: M
authors.initials: M
authors.lastname: Pirio
authors.forename: N
authors.initials: N
authors.lastname: DeChene
authors.forename: A
authors.initials: A
authors.lastname: Dafforn
authors.forename: S
authors.initials: S
authors.lastname: Rose
mesh_terms: D002692:Chlamydia trachomatis / Q000235:genetics; D004247:DNA / Q000032:analysis*; D004269:DNA, Bacterial / Q000032:analysis; D007840:Latex / Q000737:chemistry; D008163:Luminescent Measurements; D009169:Mycobacterium tuberculosis / Q000235:genetics; D009841:Oligonucleotides / Q000737:chemistry; D016133:Polymerase Chain Reaction
chemical_list: D004269:DNA, Bacterial; D007840:Latex; D009841:Oligonucleotides; D004247:DNA