Label free and amplified detection of cancer marker EBNA-1 by DNA probe based biosensors

Biosens Bioelectron. 2011 Dec 15;30(1):272-5. doi: 10.1016/j.bios.2011.09.025. Epub 2011 Sep 29.

Abstract

Epstein-Barr virus (EBV) is a human herpes virus that has been associated with several malignancies as Burkitt's lymphoma, nasopharyngeal carcinoma and Hodgkin's disease. All EBV associated malignancies showed a distinct viral gene expression pattern, while Epstein-Barr nuclear antigen 1 (EBNA-1) is constitutively expressed in all such disorders. Here, the development of a biosensor to detect EBNA-1 protein is reported, which was based on a nucleic acid bioreceptor and a quartz crystal microbalance with a dissipation monitoring (QCM-D) transducer. The DNA probe for EBNA-1 detection was designed and synthesized to mimic its palindromic target sites in the EBV genome. This DNA probe was immobilized on the Au-surface of a QCM-D electrode, followed by the blocking of the accessible Au-surface with 6-mercapto-1-hexanol (6-MHO). The system showed a limit of detection of 50 ng/mL in direct detection of EBNA-1, however, the sensitivity was improved by 2 orders of magnitude (0.5 ng/mL) when an amplification cascade, employing antibodies labeled with alkaline phosphatase (AP), was applied to the system.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomarkers, Tumor / analysis*
  • Biosensing Techniques / instrumentation*
  • DNA Probes / chemistry*
  • DNA Probes / genetics
  • Epstein-Barr Virus Nuclear Antigens / analysis*
  • Equipment Design
  • Equipment Failure Analysis
  • Micro-Electrical-Mechanical Systems / instrumentation*
  • Molecular Probe Techniques / instrumentation*
  • Nucleic Acid Amplification Techniques / instrumentation*
  • Staining and Labeling

Substances

  • Biomarkers, Tumor
  • DNA Probes
  • Epstein-Barr Virus Nuclear Antigens
  • EBV-encoded nuclear antigen 1