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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Fang, Xiangyi Yuan, Lu Yang, Yucong Yang, Fan Liang, Xiaohui |
| Description | Author Affiliation: Yang Y ( Department of Clinical Laboratory, the First Affiliated Hospital of Medical College, Xi'an Jiaotong University, Xi'an, PR China.) |
| 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. |
| ISSN | 09565663 |
| Volume Number | 46 |
| e-ISSN | 18734235 |
| Journal | Biosensors and Bioelectronics |
| Language | English |
| Publisher | Elsevier |
| Publisher Date | 2013-08-15 |
| Publisher Place | Great Britain (UK) |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Dna Genetics Hla-b27 Antigen Surface Plasmon Resonance Methods Base Sequence Analysis Blood Humans Limit Of Detection Nucleic Acid Hybridization Oligonucleotide Array Sequence Analysis Polymerase Chain Reaction Spondylitis, Ankylosing Diagnosis Evaluation Studies Journal Article Research Support, Non-u.s. Gov't Discipline Biotechnology |
| Content Type | Text |
| Resource Type | Article |
| Subject | Nanoscience and Nanotechnology Medicine Biophysics Biomedical Engineering Biotechnology Electrochemistry |
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