Deployable, field-sustainable, reverse transcription-polymerase chain reaction assays for rapid screening and serotype identification of dengue virus in mosquitoes.
Dengue virus universal and serotype 1 to 4 fluorogenic probe hydrolysis, reverse transcription (RT)-polymerase chain reaction (PCR) assays and positive-control RNA template were freeze-dried in a thermally stable, hydrolytic enzyme-resistant format and deployed for testing in a dengue fever-endemic...
| Publicado en: | Military Medicine Vol. 172; no. 3; pp. 329 - 335 |
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| Autores principales: | , , , , , , , , , , , |
| Formato: | research Journal Article |
| Publicado: |
Oxford University Press / USA
Mar2007
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| Acceso en línea: | Ver este registro en EBSCOhost |
| fields | @attributes: recordID: 1 pdfLink: plink: https://search.ebscohost.com/login.aspx?direct=true&db=ccm&AN=105986270&site=ehost-live header: @attributes: shortDbName: ccm uiTerm: 105986270 longDbName: CINAHL Complete uiTag: AN controlInfo: bkinfo: dissinfo: jinfo: jid: 00264075 4DV jtl: Military Medicine issn: 00264075 maglogo: N pubinfo: dt: Mar2007 vid: 172 iid: 3 pid: 622 pub: Oxford University Press / USA artinfo: ui: 105986270 105986270 NLM17436782 2009545653 10.7205/milmed.172.3.329 NLM17436782 105986270 ppf: 329 ppct: 6 formats: fmt: @attributes: type: P tig: atl: Deployable, field-sustainable, reverse transcription-polymerase chain reaction assays for rapid screening and serotype identification of dengue virus in mosquitoes. aug: au: McAvin JC Powers MD Blow JA Putnam JL Huff WB Swaby JA McAvin, James C Powers, Michael D Blow, Jamie A Putnam, John L Huff, William B Swaby, James A affil: U.S. Air Force Institute for Operational Health, Brooks City-Base, San Antonio, TX 78235-5237, USA sug: subj: Flaviviridae Health Screening Mobile Health Units Mosquitoes Reverse Transcriptase Polymerase Chain Reaction Equipment and Supplies Animals Female Male Pilot Studies Reverse Transcriptase Polymerase Chain Reaction Methods RNA Thailand Time Factors Viruses Animal Studies Female Male ab: Dengue virus universal and serotype 1 to 4 fluorogenic probe hydrolysis, reverse transcription (RT)-polymerase chain reaction (PCR) assays and positive-control RNA template were freeze-dried in a thermally stable, hydrolytic enzyme-resistant format and deployed for testing in a dengue fever-endemic region of Thailand. The study site presented austere testing conditions. Field-collected Aedes aegypti mosquitoes spiked with inoculated A. aegypti mosquitoes and individual and pooled, field-collected, A. aegypti, A. albopictus, and Culex tritaeniorhynchus mosquitoes were used for RT-PCR assay evaluations. For dengue virus-inoculated A. aegypti mosquitoes and spiked samples, in vitro sensitivity and specificity results for all five assays were concordant with indirect fluorescent antibody assay results. A single pool of field-collected, female, A. aegypti mosquitoes was identified as dengue virus positive. Cross-reactivity was not observed across heterologous serotypes, mosquito vectors, or human DNA. The limit of detection was >7 to < or =70 genomic equivalents. Sample processing and analysis required <2 hours. These results show promise of field-formatted RT-PCR reagents for rapid, sensitive, specific dengue virus screening and serotype identification in mosquitoes under field-deployed conditions. pubtype: Academic Journal doctype: research Journal Article ougenre: Article language: English refInfo: holdings: @attributes: islocal: N |
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