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Titolo:
DETECTION OF DENGUE-2 VIRAL-RNA BY REVERSIBLE TARGET CAPTURE HYBRIDIZATION
Autore:
CHANDLER LJ; BLAIR CD; BEATY BJ;
Indirizzi:
COLORADO STATE UNIV,COLL VET MED & BIOMED SCI,DEPT MICROBIOL,ARTHROPOD BORNE & INFECT DIS LAB FT COLLINS CO 80523 COLORADO STATE UNIV,COLL VET MED & BIOMED SCI,DEPT MICROBIOL,ARTHROPOD BORNE & INFECT DIS LAB FT COLLINS CO 80523
Titolo Testata:
Journal of clinical microbiology
fascicolo: 10, volume: 31, anno: 1993,
pagine: 2641 - 2647
SICI:
0095-1137(1993)31:10<2641:DODVBR>2.0.ZU;2-O
Fonte:
ISI
Lingua:
ENG
Soggetto:
NUCLEIC-ACID HYBRIDIZATION; POLYMERASE CHAIN-REACTION; AEDES-ALBOPICTUS; RAPID DETECTION; VIRUS TYPE-2; PROBES; AMPLIFICATION;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
18
Recensione:
Indirizzi per estratti:
Citazione:
L.J. Chandler et al., "DETECTION OF DENGUE-2 VIRAL-RNA BY REVERSIBLE TARGET CAPTURE HYBRIDIZATION", Journal of clinical microbiology, 31(10), 1993, pp. 2641-2647

Abstract

A reversible target capture (RTC) sandwich hybridization technique has been developed for the detection of dengue-2 viral RNA. The RTC is aform of sandwich hybridization that utilizes two probes: a poly(dA)-tailed capture probe and a labeled detector probe. Following hybridization of both probes to the analyte in solution, the poly(dA)-tailed capture probe is used to selectively remove the hybrids by capture on oligo(dT)-coated paramagnetic beads. After elution from the beads, the presence of specific hybrids is revealed by detection of the labeled probe. After optimization of all parameters by using P-32-labeled probes,digoxigenin was used as a label to preclude the use of radioisotopes. The sensitivity of the developed RTC procedure was determined. The lowest amount of virus detectable in cell culture lysates by using P-32-labeled probes was 20 PFU, while with digoxigenin-labeled probes, 200 PFU was detectable. The RTC procedure also detected dengue-2 virus in infected mosquitoes, both individually and in pools. The RTC has the advantage of being performed directly on crude samples, eliminating theneed for phenol extraction and purification of target nucleic acids. These results indicate that the RTC procedure is sensitive, rapid, andeasy to perform and that its use in surveillance programs will allow detection of dengue virus in pools of mosquitoes more rapidly than current procedures.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 02/12/20 alle ore 17:54:37