A DNA sensor based on surface plasmon resonance for apoptosis-associated genes detection.

A practical and simple DNA sensor based on surface plasmon resonance (SPR) had been developed to determine apoptosis-associated genes, bcl-2 and bax. This SPR sensor was designed on the basis of simultaneous multi-wavelength detection. The complementary sequences of bcl-2 or bax oligonucleotide labeled with biotin were used as the probes. Biotin-avidin system was used to immobilize the bio-DNA on the sensor surface. The assembling processes and conditions for the DNA sensor were examined. The SPR sensor could be used to monitor the process of the immobility of the bio-DNA and DNA hybridization in real-time. The determination range of bcl-2 and bax oligonucleotide (20 bases) were 50-400 ng/mL. The determination range of polymerase chain reaction (PCR) product of bcl-2 (405 bases) was 5-60 ng/mL and PCR product of bax (538 bases) was 5-40 ng/mL. The stability, reversibility and specificity of the DNA sensor were also investigated. It was found from the experiment that the sensor could be applied for a quite long time (about 90 times). The relative standard deviation (R.S.D.) for determination oligonucleotides and PCR products of bcl-2 were 1.2 and 1.3%, respectively. The interference of noncomplementary DNA sequence with the determination of DNA was examined and it was found that noncomplementary 20-mer and 21-mer DNA (p53 and p21) do not affect the determination of bcl-2 or bax. This device could be used to study apoptosis and signal transduction routine genes. The sensor was shown to be of simplicity, sensitivity, selectivity, rapid response and cost effectiveness.

[1]  Ying Mu,et al.  A novel sandwich immunosensing method for measuring cardiac troponin I in sera. , 2003, Analytical biochemistry.

[2]  Chris A. Rowe-Taitt,et al.  Array biosensor for detection of biohazards. , 2000, Biosensors & bioelectronics.

[3]  M. Clarke,et al.  The Bcl-2 family of proteins: regulators of cell death and survival. , 1994, Trends in cell biology.

[4]  Xiao-yong Lei,et al.  Bcl-2 small interfering RNA sensitizes cisplatin-resistant human lung adenocarcinoma A549/DDP cell to cisplatin and diallyl disulfide. , 2007, Acta biochimica et biophysica Sinica.

[5]  Avraham Rasooly,et al.  Spectral surface plasmon resonance biosensor for detection of staphylococcal enterotoxin B in milk. , 2002, International journal of food microbiology.

[6]  M. Thompson,et al.  Genosensor technology and the detention of interfacial nucleic acid chemistry , 1997 .

[7]  F. van Knapen,et al.  Surface plasmon resonance (BIACORE) detection of serum antibodies against Salmonella enteritidis and Salmonella typhimurium. , 2002, Journal of immunological methods.

[8]  S. Korsmeyer,et al.  Molecular thanatopsis: a discourse on the BCL2 family and cell death. , 1996, Blood.

[9]  R. Schreiber,et al.  Bcl-2 is an inner mitochondrial membrane protein that blocks programmed cell death , 1990, Nature.

[10]  F. Caruso,et al.  Quartz crystal microbalance study of DNA immobilization and hybridization for nucleic Acid sensor development. , 1997, Analytical chemistry.

[11]  H. Perlman,et al.  An elevated bax/bcl-2 ratio corresponds with the onset of prostate epithelial cell apoptosis , 1999, Cell Death and Differentiation.

[12]  Shusheng Zhang,et al.  Disposable biosensor based on enzyme immobilized on Au-chitosan-modified indium tin oxide electrode with flow injection amperometric analysis. , 2007, Analytical biochemistry.

[13]  S. Agrawal,et al.  Efficient methods for attaching non-radioactive labels to the 5' ends of synthetic oligodeoxyribonucleotides. , 1986, Nucleic acids research.

[14]  R. Craig.  The bcl-2 gene family. , 1995, Seminars in cancer biology.

[15]  S. Korsmeyer,et al.  Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programed cell death , 1993, Cell.