title: Detection of HLA-B*27 gene using a spectral plasmon resonance imaging system.
abstract: Detection of HLA-B*27 gene is clinically important due to its strong association with diseases, such as ankylosing spondylitis. Nucleic acid-based biosensors represent a promising clinical tool for gene diagnosis. Surface plasmon resonance (SPR) can be used to monitor DNA-DNA hybridization in real-time and without any prior labeling step. Here, a self-built HLA-B*27 positive PCR products spectral SPR imaging system was used for multichannel direct-detection of a specific sequence of the HLA-B*27 gene. Thiol-labeled single-stranded oligonucleotide probes, which were proved to be superior to amine-labeled probes in immobilization, were immobilized onto the surfaces of the gold spots on the sensor chip to target the specific sequence in the HLA-B*27 gene in blood. SPR measurements were performed with different concentrations of synthetic target DNA sequence. The calibration curve of synthetic target sequence showed a good linear relationship between the concentration of the synthetic target sequence and the shift of the SPR wavelength from 10 nM to 500 nM with a detection limit of 5 nM. The HLA-B*27 gene was isolated from human whole blood and amplified using polymerase chain reaction (PCR). PCR products were measured using the SPR imaging system. HLA-B*27 positive PCR products showed significant SPR wavelength shift, while synthetic non-complementary sequence and negative PCR products showed no significant SPR wavelength shift. The method is high-specific, high-throughput and label-free.
authors.affiliation: Department of Clinical Laboratory, the First Affiliated Hospital of Medical College, Xi'an Jiaotong University, Xi'an, PR China.
authors.forename: Yucong
authors.initials: Y
authors.lastname: Yang
authors.forename: Lu
authors.initials: L
authors.lastname: Yuan
authors.forename: Xiangyi
authors.initials: X
authors.lastname: Fang
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Liang
authors.forename: Fan
authors.initials: F
authors.lastname: Yang
mesh_terms: D001483:Base Sequence; D004247:DNA / Q000032:analysis / Q000097:blood / Q000235:genetics*; D015796:HLA-B27 Antigen / Q000235:genetics*; D006801:Humans; D057230:Limit of Detection; D009693:Nucleic Acid Hybridization / Q000379:methods; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D016133:Polymerase Chain Reaction; D013167:Spondylitis, Ankylosing / Q000097:blood / Q000175:diagnosis; D020349:Surface Plasmon Resonance / Q000379:methods*
chemical_list: D015796:HLA-B27 Antigen; D004247:DNA