Label free and amplified detection of cancer marker EBNA-1 by DNA probe based biosensors.
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.
Biofunctional Nanomaterials Department, CIC biomaGUNE, Donostia, San Sebastian, 20009, Spain.
Gaizka
G
Garai-Ibabe
Ruta
R
Grinyte
Efim I
EI
Golub
Allon
A
Canaan
Marc Lamy
ML
de la Chapelle
Robert S
RS
Marks
Valeri
V
Pavlov
D014408:Biomarkers, Tumor / Q000032:analysis*; D015374:Biosensing Techniques / Q000295:instrumentation*; D015342:DNA Probes / Q000737:chemistry* / Q000235:genetics; D019309:Epstein-Barr Virus Nuclear Antigens / Q000032:analysis*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D055617:Micro-Electrical-Mechanical Systems / Q000295:instrumentation*; D015336:Molecular Probe Techniques / Q000295:instrumentation*; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation*; D013194:Staining and Labeling
D014408:Biomarkers, Tumor; D015342:DNA Probes; D019309:Epstein-Barr Virus Nuclear Antigens; C417029:EBV-encoded nuclear antigen 1