Evaluation of a field-portable DNA microarray platform and nucleic acid amplification strategies for the detection of arboviruses, arthropods, and bloodmeals.
Abstract: Highly multiplexed assays, such as microarrays, can benefit arbovirus surveillance by allowing researchers to screen for hundreds of targets at once. We evaluated amplification strategies and the practicality of a portable DNA microarray platform to analyze virus-infected mosquitoes. The prototype microarray design used here targeted the non-structural protein 5, ribosomal RNA, and cytochrome b genes for the detection of flaviviruses, mosquitoes, and bloodmeals, respectively. We identified 13 of 14 flaviviruses from virus inoculated mosquitoes and cultured cells. Additionally, we differentiated between four mosquito genera and eight whole blood samples. The microarray platform was field evaluated in Thailand and successfully identified flaviviruses (Culex flavivirus, dengue-3, and Japanese encephalitis viruses), differentiated between mosquito genera (Aedes, Armigeres, Culex, and Mansonia), and detected mammalian bloodmeals (human and dog). We showed that the microarray platform and amplification strategies described here can be used to discern specific information on a wide variety of viruses and their vectors.
Publication Date: 2012-12-18 PubMed ID: 23249687PubMed Central: PMC3583313DOI: 10.4269/ajtmh.2012.12-0048Google Scholar: Lookup
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- Evaluation Study
- Journal Article
- Research Support
- U.S. Gov't
- Non-P.H.S.
Summary
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The research article revolves around the testing of a portable DNA microarray platform and the effectiveness of strategies used to amplify nucleic acids. The tests successfully detected the presence of several species of arboviruses, arthropods, and sources of bloodmeals.
Research Objective
- The main focus of the study was to analyze the effectiveness and practicality of a DNA microarray platform that is portable. This platform was employed to detect flaviviruses, arthropods and bloodmeals. The tests were done by scrutinizing mosquitoes that were virus-infected.
Nucleic Acid Amplification Strategies
- The study used different amplification strategies, an integral part of microarray tests. Amplification strategies play a vital role in improving the sensitivity of the tests and increase chances of detecting trace amounts of genetic material. The goal here was to evaluate the impact of these strategies on the detection of arboviruses, arthropods, and bloodmeals.
Microarray Platform Evaluation
- The microarray platform was designed to target ribosomal RNA, non-structural protein 5, and cytochrome b genes for the detection of mosquitoes, flaviviruses, and bloodmeals respectively.
- The platform was able to effectively identify 13 out of 14 flaviviruses from virus-inoculated mosquitoes and cultured cells.
- The platform also successfully distinguished between four mosquito genera and showed capability in identifying eight different whole blood samples.
- A field evaluation in Thailand successfully identified key flaviviruses, differentiated between mosquito genera and detected mammalian bloodmeals like human and dog.
Implication of the Findings
- The successful detection and differentiation of multiple species by the portable DNA microarray platform substantiates the potential of the device for use in virus and vector identification.
- The amplifier strategies showcased effectiveness in improving the device’s sensitivity and thus, open up possibilities for improvements in detection techniques of several blood-borne diseases.
Cite This Article
APA
Grubaugh ND, Petz LN, Melanson VR, McMenamy SS, Turell MJ, Long LS, Pisarcik SE, Kengluecha A, Jaichapor B, O'Guinn ML, Lee JS.
(2012).
Evaluation of a field-portable DNA microarray platform and nucleic acid amplification strategies for the detection of arboviruses, arthropods, and bloodmeals.
Am J Trop Med Hyg, 88(2), 245-253.
https://doi.org/10.4269/ajtmh.2012.12-0048 Publication
Researcher Affiliations
- Virology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD 21702, USA. nathan.grubaugh@colostate.edu
MeSH Terms
- Animals
- Arboviruses / genetics
- Arboviruses / isolation & purification
- Arboviruses / pathogenicity
- Arthropods / virology
- Blood / virology
- Computational Biology
- Culicidae / virology
- Cytochromes b / genetics
- DNA, Viral / genetics
- Dogs
- Equidae
- Flavivirus / genetics
- Flavivirus / isolation & purification
- Flavivirus / pathogenicity
- Genes, Viral
- Horses
- Humans
- Insect Vectors / virology
- Oligonucleotide Array Sequence Analysis / methods
- RNA, Viral / genetics
- Real-Time Polymerase Chain Reaction
- Thailand
- Viral Nonstructural Proteins / genetics
- Viral Nonstructural Proteins / metabolism
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