TY - JOUR
T1 - Rapid and amplification-free detection of fish pathogens by utilizing a molecular beacon-based microfluidic system
AU - Su, Yi Chih
AU - Wang, Chih Hung
AU - Chang, Wen Hsin
AU - Chen, Tzong Yueh
AU - Lee, Gwo Bin
N1 - Funding Information:
The authors gratefully acknowledge the financial support provided to this study by the National Science Council in Taiwan ( NSC102-2218-E-007-001 ). The authors would like to thank Dr. Kuangwen Hsieh for editing and providing valuable feedback. The authors also thank Dr. Chih-Peng Chang for providing some virus samples.
PY - 2015/1/15
Y1 - 2015/1/15
N2 - Nervous necrosis virus (NNV) and iridovirus are highly infectious pathogens that can cause lethal diseases in various species of fish. These infectious diseases have no effective treatments and the mortality rate is over 80%, which could cause dramatic economic losses in the aquaculture industry. Conventional diagnostic methods of NNV or iridovirus infected fishes, such as virus culture, enzyme-linked immunosorbent assays and nucleic acid assays usually require time-consuming and complex procedures performed by specialized technicians with delicate laboratory facilities. Rapid, simple, accurate and on-site detection of NNV and iridovirus infections would enable timely preventive measures such as immediate sacrifice of infected fishes, and is therefore critically needed for the aquaculture industry. In this study, a microfluidic-based assay that employ magnetic beads conjugated with viral deoxyribonucleic acid (DNA) capturing probes and fluorescent DNA molecular beacons were developed to rapidly detect NNV and iridovirus. Importantly, this new assay was realized in an integrated microfluidic system with a custom-made control system. With this approach, direct and automated NNV and iridovirus detection from infected fishes can be achieved in less than 30. min. Therefore, this molecular-beacon based microfluidic system presents a potentially promising tool for rapid diagnosis of fish pathogens in the field in the future.
AB - Nervous necrosis virus (NNV) and iridovirus are highly infectious pathogens that can cause lethal diseases in various species of fish. These infectious diseases have no effective treatments and the mortality rate is over 80%, which could cause dramatic economic losses in the aquaculture industry. Conventional diagnostic methods of NNV or iridovirus infected fishes, such as virus culture, enzyme-linked immunosorbent assays and nucleic acid assays usually require time-consuming and complex procedures performed by specialized technicians with delicate laboratory facilities. Rapid, simple, accurate and on-site detection of NNV and iridovirus infections would enable timely preventive measures such as immediate sacrifice of infected fishes, and is therefore critically needed for the aquaculture industry. In this study, a microfluidic-based assay that employ magnetic beads conjugated with viral deoxyribonucleic acid (DNA) capturing probes and fluorescent DNA molecular beacons were developed to rapidly detect NNV and iridovirus. Importantly, this new assay was realized in an integrated microfluidic system with a custom-made control system. With this approach, direct and automated NNV and iridovirus detection from infected fishes can be achieved in less than 30. min. Therefore, this molecular-beacon based microfluidic system presents a potentially promising tool for rapid diagnosis of fish pathogens in the field in the future.
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U2 - 10.1016/j.bios.2014.07.035
DO - 10.1016/j.bios.2014.07.035
M3 - Article
C2 - 25089817
AN - SCOPUS:84905245405
SN - 0956-5663
VL - 63
SP - 196
EP - 203
JO - Biosensors and Bioelectronics
JF - Biosensors and Bioelectronics
ER -
