Format

Send to

Choose Destination
Biosens Bioelectron. 2015 Jan 15;63:196-203. doi: 10.1016/j.bios.2014.07.035. Epub 2014 Jul 24.

Rapid and amplification-free detection of fish pathogens by utilizing a molecular beacon-based microfluidic system.

Author information

1
Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu 30013, Taiwan.
2
Institute of Biotechnology, National Cheng Kung University, Tainan 70101, Taiwan. Electronic address: ibcty@mail.ncku.edu.tw.
3
Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu 30013, Taiwan; Institute of Biomedical Engineering, National Tsing Hua University, Hsinchu 30013, Taiwan; Institute of NanoEngineering and Microsystems, National Tsing Hua University, Hsinchu 30013, Taiwan. Electronic address: gwobin@pme.nthu.edu.tw.

Abstract

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.

KEYWORDS:

Iridovirus; Microfluidics; Molecular beacon; Nervous necrosis virus; Probe conjugated magnetic beads

PMID:
25089817
DOI:
10.1016/j.bios.2014.07.035
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Elsevier Science
Loading ...
Support Center