Trace Determination of Grouper Nervous Necrosis Virus in Contaminated Larvae and Pond Water Samples Using Label-Free Fiber Optic Nanoplasmonic Biosensor

• Published in: Biosensors (Basel) Vol. 12; no. 10; p. 907
• Main Authors: Chen, Yuan-Yu, Wu, Chih-Lu, Hsu, Chia-Wei, Wang, Chih-Hui, Su, Chung-Rui, Huang, Chun-Jen, Chen, Hau-Ren, Chau, Lai-Kwan, Wang, Shau-Chun
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 01-10-2022; MDPI
• Subjects: Acids; Animals; Antibodies; Aquaculture; Aquaculture facilities; Bass; Biosensing Techniques; Biosensors; Calibration; Coat protein; Diagnosis; Diseases; Enzyme-linked immunosorbent assay; Epidemics; Equipment and supplies; Fatalities; fiber optic particle plasmon resonance biosensor; Fiber optics; Fish; Fish Diseases - diagnosis; Fishes; Gold; gold nanoparticles; Groupers; Health aspects; Immunosensors; Immunosorbents; Larva; Larvae; larvae homogenate; Methods; Nanoparticles; Nanotechnology; Necrosis; nervous necrosis virus; Nodaviridae; on-site testing; Phosphates; Polymerase chain reaction; pond water; Ponds; Proteins; RNA polymerase; RNA virus infections; Surface plasmon resonance sensors; Surveillance; Taiwan; Viruses; Water; Water analysis; Water pollution; Water sampling; Taiwan; United States > US; larvae homogenate; on-site testing; nervous necrosis virus; fiber optic particle plasmon resonance biosensor; gold nanoparticles; pond water
• ISSN: 2079-6374; 2079-6374
• DOI: 10.3390/bios12100907

We developed a fast (<20 min), label-free fiber optic particle plasmon resonance (FOPPR) immunosensing method to detect nervous necrosis virus (NNV), which often infects high-value economic aquatic species, such as grouper. Using spiked NNV particles in a phosphate buffer as samples, the standard calibration curve obtained was linear (R2 = 0.99) and the limit of detection (LOD) achieved was 2.75 × 104 TCID50/mL, which is superior to that obtained using enzyme-linked immunosorbent assay (ELISA). By using an enhancement method called fiber optic nanogold-linked immunosorbent assay (FONLISA), the LOD can be further improved to <1 TCID50/mL, which is comparable to that found by the conventional qPCR method. Employing the larvae homogenate samples of NNV-infected grouper, the results obtained by the FOPPR biosensor agree with those obtained by the quantitative polymerase chain reaction (qPCR) method. We also examined pond water samples from an infected container in an indoor aquaculture facility. The lowest detectable level of NNV coat protein was found to be 0.17 μg/mL, which is one order lower than the LOD reported by ELISA. Therefore, we demonstrated the potential of the FOPPR biosensor as an outbreak surveillance tool, which is able to give warning indication even when the trend of larvae death toll increment is still not clear.