Journal
BIOSENSORS & BIOELECTRONICS
Volume 86, Issue -, Pages 1054-1060Publisher
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2016.07.100
Keywords
Label-free immunoassay; Excessively tilted fiber grating (Ex-TFG); Porcine circovirus type 2 (PCV2); Staphylococcal protein A (SPA); Monoclonal antibody (MAb)
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Funding
- National Natural Science Foundation of China [61505017, 61327004, 61421002, 51276209]
- social undertakings and Livelihood security of Chongqing City [cstc2015shmszx80027, cstc2016shmszx0076]
- Chongqing University outstanding achievements transformation fund [KJZH14212]
- Foundation and cutting-edge Research Projects of Chongqing City [cstc2014jcyjA0081, cstc2013jcyjA10068]
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Using excessively tilted fiber grating (Ex-TFG) inscribed in standard single mode fiber, we developed a novel label-free immunoassay for specific detection of porcine circovirus type 2 (PCV2), which is a minim animal virus. Staphylococcal protein A (SPA) was used to modify the silanized fiber surface thus forming a SPA layer, which would greatly enhance the proportion of anti-PCV2 monoclonal antibody (MAb) bioactivity, thus improving the effectiveness of specific adsorption and binding events between anti-PCV2 MAbs and PCV2 antigens. Immunoassay experiments were carried out by monitoring the resonance wavelength shift of the proposed sensor under different PCV2 titer levels. Anti-PCV2 MAbs were thoroughly dissociated from the SPA layer by treatment with urea, and recombined to the SPA layer on the sensor surface for repeated immunoassay of PCV2. The specificity of the immunosensor was inspected by detecting porcine reproductive and respiratory syndrome virus (PRRSV) first, and PCV2 subsequently. The results showed a limit of detection (LOD) for the PCV2 immunosensor of similar to 9.371TCID(50)/mL, for a saturation value of similar to 4.801 x 10(3)TCID(50)/mL, with good repeatability and excellent specificity. (C) 2016 Elsevier B.V. All rights reserved.
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