Evaluation of a Field-Portable DNA Microarray Platform and Nucleic Acid Amplification Strategies for the Detection of Arboviruses, Arthropods, and Bloodmeals

Nathan D. Grubaugh Virology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, Maryland; Department of Entomology, United States Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand

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Lawrence N. Petz Virology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, Maryland; Department of Entomology, United States Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand

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Vanessa R. Melanson Virology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, Maryland; Department of Entomology, United States Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand

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Scott S. McMenamy Virology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, Maryland; Department of Entomology, United States Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand

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Michael J. Turell Virology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, Maryland; Department of Entomology, United States Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand

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Lewis S. Long Virology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, Maryland; Department of Entomology, United States Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand

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Sarah E. Pisarcik Virology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, Maryland; Department of Entomology, United States Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand

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Ampornpan Kengluecha Virology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, Maryland; Department of Entomology, United States Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand

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Boonsong Jaichapor Virology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, Maryland; Department of Entomology, United States Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand

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Monica L. O'Guinn Virology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, Maryland; Department of Entomology, United States Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand

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John S. Lee Virology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, Maryland; Department of Entomology, United States Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand

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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.

Author Notes

* Address correspondence to John S. Lee, Military Infectious Diseases Research Program, U.S. Army Medical Research and Materiel Command, Building 722, 504 Scott Avenue, Fort Detrick, Frederick, MD 21702. E-mail: john.s.lee@us.army.mil

Financial support: This research was funded by the Military Infectious Disease Research Program project no. U0176_09_RD and supported in part by an appointment to the Postgraduate Research Participation Program at USAMRIID administered by the Oak Ridge Institute for Science and Education through an interagency agreement between the U.S. Department of Energy and U.S. Army Medical Research and Material Command.

Authors' addresses: Nathan D. Grubaugh, Department of Microbiology, Immunology, and Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO, E-mail: nathan.grubaugh@colostate.edu. Lawrence N. Petz, Battlefield Pain Management Research Program, United States Army Institute of Surgical Research, Fort Sam Houston, TX, E-mail: lawrence.petz@us.army.mil. Vanessa R. Melanson and Lewis S. Long, Entomology Branch, Walter Reed Army Institute of Research, Silver Spring, MD, E-mails: vanessa.melanson@us.army.mil and lewis.s.long@us.army.mil. Scott S. McMenamy, Michael J. Turell, and Sarah E. Pisarcik, Virology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, MD, E-mails: scott.mcmenamy@us.army.mil, michael.j.turell@us.army.mil., and sarah.pisarcik@us.army.mil. Ampornpan Kengluecha and Boonsong Jaichapor, Department of Entomology, United States Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand, E-mails: ampornpank@afrims.org and boonsongj@afrims.org. Monica L. O'Guinn and John S. Lee, Military Infectious Diseases Research Program, U.S. Army Medical Research and Materiel Command, E-mails: monica.oguinn@us.army.mil and john.s.lee@us.army.mil.

Reprint requests: John S. Lee, Military Infectious Diseases Research Program, U.S. Army Medical Research and Materiel Command, Building 722, 504 Scott Avenue, Fort Detrick, Frederick, MD 21702, Tel: 301-619-7797, E-mail: john.s.lee@us.army.mil.

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