title: Development of a multiplexed PCR-coupled liquid bead array assay for vascular endothelial growth factor (VEGF) splice variants.
abstract: OBJECTIVE
  To develop a multiplex PCR-coupled liquid bead array assay for the expression of VEGF splice variants.
  
  
  METHODS
  The assay was based on the combination of multiplex PCR with liquid bead array technology, and optimized and evaluated in terms of analytical sensitivity, specificity, and reproducibility using the MCF-7 cell line. Clinical performance was evaluated in 16 pairs of fresh frozen cancerous and corresponding noncancerous adjacent tissues from NSCLC patients.
  
  
  RESULTS
  The assay is highly sensitive, reproducible and can detect specifically VEGF splice variants in clinical samples. When applied in 32 clinical samples it gave comparable results to RT-qPCR (concordance of 81%, 75%, 88% and 81% for PBGD, VEGF(121), VEGF(165), and VEGF(189) respectively).
  
  
  CONCLUSIONS
  The developed assay can simultaneously detect three VEGF splice variants with high specificity and sensitivity and can be further used to evaluate the role of each individual VEGF splice variant in molecular therapies targeted against VEGF.
authors.affiliation: Laboratory of Analytical Chemistry, Department of Chemistry, University of Athens, 15771, Athens, Greece.
authors.forename: C
authors.initials: C
authors.lastname: Parisi
authors.forename: A
authors.initials: A
authors.lastname: Markou
authors.forename: E S
authors.initials: ES
authors.lastname: Lianidou
mesh_terms: D002289:Carcinoma, Non-Small-Cell Lung / Q000378:metabolism; D045744:Cell Line, Tumor; D006801:Humans; D008168:Lung / Q000378:metabolism; D008175:Lung Neoplasms / Q000378:metabolism; D016555:Matched-Pair Analysis; D008863:Microspheres; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D012015:Reference Standards; D015203:Reproducibility of Results; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics* / Q000378:metabolism
chemical_list: D020033:Protein Isoforms; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A