Detection of Dengue Virus in Mosquito Extracts and Human Clinical Samples Using a Field Expedient Molecular Platform.
Dengue fever occurs in localized outbreaks and can significantly erode troop strength and mission readiness. Timely identification of dengue virus (DENV) provides for rapid and appropriate patient management decisions, such as medical evacuation and supportive therapies, as well as help to promote Force Health Protection through vector control and personal protective measures. The "Ruggedized" Advanced Pathogen Identification Device is a field-friendly PCR (Polymerase Chain Reaction) platform that can be used to facilitate early identification of DENV. We developed a dry-format PCR assay on this platform. The assay demonstrated 100% analytical specificity for detecting dengue using a cross-reactivity panel. We used a panel of 102 acute, DENV isolation positive serum samples and 25 DENV negative samples; the assay demonstrated a clinical sensitivity of 97.1% (95% C.I. 91.6-99.4%) and specificity of 96.0% (95% C.I. 79.7-99.9%) in identifying patients with dengue infection. We also used the assay to test mosquito homogenates from 28 adult female Aedes aegypti. A single DENV infected mosquito was identified using the PCR assay and confirmed using immunofluorescence as a reference method. Much of the testing was performed under austere field conditions. Together, our results demonstrate the utility of this assay for detecting DENV in vector and human samples in field environments.
Viral and Rickettsial Diseases Department, Naval Medical Research Center, 503 Robert Grant Avenue, Silver Spring, MD 20910.
Subhamoy
S
Pal
Department of Entomology, Armed Forces Research Institute of Medical Sciences, 315/6 Rajvithi Road, Bangkok 10400, Thailand.
Jason H
JH
Richardson
Department of Entomology, Armed Forces Research Institute of Medical Sciences, 315/6 Rajvithi Road, Bangkok 10400, Thailand.
Jittawadee R
JR
Murphy
Department of Entomology, Armed Forces Research Institute of Medical Sciences, 315/6 Rajvithi Road, Bangkok 10400, Thailand.
Panadda
P
Krairojananan
Department of Entomology, Armed Forces Research Institute of Medical Sciences, 315/6 Rajvithi Road, Bangkok 10400, Thailand.
Patcharee
P
Kongtak
Department of Entomology, Armed Forces Research Institute of Medical Sciences, 315/6 Rajvithi Road, Bangkok 10400, Thailand.
Boonsong
B
Jaichapor
Department of Entomology, Armed Forces Research Institute of Medical Sciences, 315/6 Rajvithi Road, Bangkok 10400, Thailand.
Prasan
P
Kankaew
Viral and Rickettsial Diseases Department, Naval Medical Research Center, 503 Robert Grant Avenue, Silver Spring, MD 20910.
Sajeewane
S
Ekanayake
Detachment 3, USAF School of Aerospace Medicine, Kadena AB, Okinawa 96368, Japan.
Timothy J
TJ
Davis
Advanced Technology Center, 3201 Sidney Brooks, San Antonio, TX 78235.
David L
DL
Maserang
BioFire Diagnostics LLC, 390 Wakara Way, Salt Lake City, UT 84108.
David H F
DH
Teng
BioFire Diagnostics LLC, 390 Wakara Way, Salt Lake City, UT 84108.
Robert J
RJ
Crisp
Viral and Rickettsial Diseases Department, Naval Medical Research Center, 503 Robert Grant Avenue, Silver Spring, MD 20910.
Shuenn-Jue L
SJ
Wu
Joint Programs Executive Office, Chemical and Biological Defense, 1504 Porter Street, Fort Detrick, MD 21702.
Russell E
RE
Coleman
Department of Entomology, Armed Forces Research Institute of Medical Sciences, 315/6 Rajvithi Road, Bangkok 10400, Thailand.
James C
JC
McAvin
Clinical Research Division, 59th Medical Wing, 2200 Bergquist Drive, Building 4430, Lackland AFB, TX 78233.
James A
JA
Swaby
D000330:Aedes / Q000821:virology*; D000818:Animals; D003715:Dengue / Q000097:blood / Q000821:virology*; D003716:Dengue Virus / Q000235:genetics / Q000302:isolation & purification*; D004199:Disease Vectors; D005260:Female; D006801:Humans; D008887:Military Medicine / Q000295:instrumentation* / Q000379:methods; D008952:Mobile Health Units; D016133:Polymerase Chain Reaction / Q000295:instrumentation* / Q000379:methods; D012680:Sensitivity and Specificity; D014481:United States