A Novel Pan-Flavivirus Detection and Identification Assay Based on RT-qPCR and Microarray.
The genus Flavivirus includes arthropod-borne viruses responsible for a large number of infections in humans and economically important animals. While RT-PCR protocols for specific detection of most Flavivirus species are available, there has been also a demand for a broad-range Flavivirus assay cov...
| Publicado en: | BioMed Research International Vol. 2017; pp. 1 - 13 |
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| Autores principales: | , , , , , , |
| Formato: | pictorial research tables/charts Journal Article |
| Publicado: |
Wiley-Blackwell
5/24/2017
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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=123209146&site=ehost-live header: @attributes: shortDbName: ccm uiTerm: 123209146 longDbName: CINAHL Complete uiTag: AN controlInfo: bkinfo: dissinfo: jinfo: jid: 23146133 FT2T jtl: BioMed Research International issn: 23146133 maglogo: N pubinfo: dt: 5/24/2017 vid: 2017 pid: 480 pub: Wiley-Blackwell place: Malden, Massachusetts artinfo: ui: 123209146 123209146 123209146 10.1155/2017/4248756 123209146 ppf: 1 ppct: 12 formats: fmt: @attributes: type: P tig: atl: A Novel Pan-Flavivirus Detection and Identification Assay Based on RT-qPCR and Microarray. aug: au: Vina-Rodriguez, Ariel Sachse, Konrad Ziegler, Ute Chaintoutis, Serafeim C. Keller, Markus Groschup, Martin H. Eiden, Martin affil: Institute for Novel and Emerging Infectious Diseases, Friedrich-Loeffler-Institut (Federal Research Institute for Animal Health), Insel Riems, Greifswald, Germany sug: subj: Flavivirus Analysis West Nile Virus Microarray Analysis Polymerase Chain Reaction Mosquito Vectors Validation Studies Sensitivity and Specificity Nucleic Acid Probes Animal Studies Mosquitoes Greece Greece Disease Surveillance ab: The genus Flavivirus includes arthropod-borne viruses responsible for a large number of infections in humans and economically important animals. While RT-PCR protocols for specific detection of most Flavivirus species are available, there has been also a demand for a broad-range Flavivirus assay covering all members of the genus. It is particularly challenging to balance specificity at genus level with equal sensitivity towards each target species. In the present study, a novel assay combining a SYBR Green-based RT-qPCR with a low-density DNA microarray has been developed. Validation experiments confirmed that the RT-qPCR exhibited roughly equal sensitivity of detection and quantification for all flaviviruses tested. These PCR products are subjected to hybridization on a microarray carrying 84 different oligonucleotide probes that represent all known Flavivirus species. This assay has been used as a screening and confirmation tool for Flavivirus presence in laboratory and field samples, and it performed successfully in international External Quality Assessment of NAT studies. Twenty-six Flavivirus strains were tested with the assay, showing equivalent or superior characteristics compared with the original or even with species-specific RT-PCRs. As an example, test results on West Nile virus detection in a panel of 340 mosquito pool samples from Greece are presented. pubtype: Academic Journal doctype: pictorial research tables/charts Journal Article ougenre: Article language: English refInfo: holdings: @attributes: islocal: N |
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