Detection of severe acute respiratory syndrome (SARS) coronavirus nucleocapsid protein in human serum using a localized surface plasmon coupled fluorescence fiber-optic biosensor

被引:106
作者
Huang, Jason C. [1 ,2 ,5 ]
Chang, Ying-Feng [3 ]
Chen, Kuan-Hsuan [1 ,6 ]
Su, Li-Chen [4 ]
Lee, Chun-Wei [2 ,6 ]
Chen, Chii-Chang [4 ]
Chen, Yi-Ming Arthur [1 ,6 ]
Chou, Chien [3 ,4 ,7 ]
机构
[1] Natl Yang Ming Univ, AIDS Prevent & Res Ctr, Taipei 112, Taiwan
[2] Natl Yang Ming Univ, Dept Biotechnol & Lab Sci Med, Taipei 112, Taiwan
[3] Natl Yang Ming Univ, Inst Biophoton, Taipei 112, Taiwan
[4] Natl Cent Univ, Dept Opt & Photon, Tao Yuan 320, Taiwan
[5] Taipei City Hosp, Dept Educ & Res, Taipei 112, Taiwan
[6] Natl Yang Ming Univ, Inst Microbiol & Immunol, Taipei 112, Taiwan
[7] Chang Gung Univ, Grad Inst Electroopt Engn, Tao Yuan 333, Taiwan
关键词
Fiber-optic biosensor; Gold nanoparticle; Localized surface plasmon; Nucleocapsid protein; Severe acute respiratory syndrome (SARS); LINKED-IMMUNOSORBENT-ASSAY; REVERSE-TRANSCRIPTASE PCR; MONOCLONAL-ANTIBODY; SENSITIVE DETECTION; STOOL SPECIMENS; SPIKE PROTEIN; THROAT WASH; ANTIGEN; AMPLIFICATION; NANOPARTICLES;
D O I
10.1016/j.bios.2009.07.012
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
In order to enhance the sensitivity of conventional immunoassay technology for the detection of SARS coronavirus (SARS-CoV) nucleocapsid protein (N protein), we developed a localized surface plasmon coupled fluorescence (LSPCF) fiber-optic biosensor that combines sandwich immunoassay with the LSP technique. Experimentally, a linear relationship between the fluorescence signal and the concentration of recombinant SARS-CoV N (GST-N) protein in buffer solution could be observed from 0.1 pg/mL to 1 ng/mL In addition, the concentration of GST-N protein in diluted serum across a similar range could also be measured. The correlation coefficients (linear scale) for these two measurements were 0.9469 and 0.9624, respectively. In comparison with conventional enzyme linked immunosorbent assay (ELISA), the detection limit of the LSPCF fiber-optic biosensor for the GST-N protein was improved at least 10(4)-fold using the same monoclonal antibodies. Therefore, the LSPCF fiber-optic biosensor shows an ability to detect very low concentration (similar to 1 pg/mL) of SARS-CoV N protein in serum. The biosensor should help with the early diagnosis of SARS infection. (C) 2009 Elsevier B.V. All rights reserved.
引用
收藏
页码:320 / 325
页数:6
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