Catalogo Articoli (Spogli Riviste)

OPAC HELP

Titolo:
Simultaneous quantitation of two orchid viruses by the TaqMan (R) real-time RT-PCR
Autore:
Eun, AJC; Seoh, ML; Wong, SM;
Indirizzi:
Natl Univ Singapore, Dept Biol Sci, Singapore 117543, Singapore Natl Univ Singapore Singapore Singapore 117543 ngapore 117543, Singapore
Titolo Testata:
JOURNAL OF VIROLOGICAL METHODS
fascicolo: 1-2, volume: 87, anno: 2000,
pagine: 151 - 160
SICI:
0166-0934(200006)87:1-2<151:SQOTOV>2.0.ZU;2-L
Fonte:
ISI
Lingua:
ENG
Soggetto:
POLYMERASE-CHAIN-REACTION; CYMBIDIUM MOSAIC POTEXVIRUS; ODONTOGLOSSUM RINGSPOT TOBAMOVIRUS; RAPID DETECTION; ASSAY; DNA; IMMUNOCAPTURE; EXPRESSION; PRODUCT; TUMORS;
Keywords:
cymbidium mosaic potexvirus; odontoglossum ringspot tobamovirus; TaqMan((R)); fluorogenic 5 ' nuclease assay; simultaneous quantification; one-tube RT-PCR;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
31
Recensione:
Indirizzi per estratti:
Indirizzo: Wong, SM Natl Univ Singapore, Dept Biol Sci, Kent Ridge, Singapore 117543,Singapore Natl Univ Singapore Kent Ridge Singapore Singapore 117543 gapore
Citazione:
A.J.C. Eun et al., "Simultaneous quantitation of two orchid viruses by the TaqMan (R) real-time RT-PCR", J VIROL MET, 87(1-2), 2000, pp. 151-160

Abstract

Simultaneous quantitation of two orchid viruses, cymbidium mosaic potexvirus (CymMV) and odontoglossum ringspot tobamovirus (ORSV), were carried out using the TaqMan((R)) real-time RT-PCR, a novel detection technique that combines RT-PCR with the power of fluorescent detection. Four TaqMan((R)) probes were synthesized, targeting at the RNA-dependent RNA polymerase (RdRp) and coat protein (CP) genes of both viruses. The reporter dye FAM (6-carboxyfluorescein) was used to label the 5' terminus of probes specific to CymMV, while TET (tetrachloro-6-carboxyfluorescein) was used for the ORSV probes. TAMRA (6-carboxy-tetramethyl-rhodamine), which was attached at the 3' terminus of each probe, was used as the universal quencher. With increasing amounts of standard RNA templates, the respective threshold cycle (C-T) values were determined and a linear relationship was established between these C-T values and the logarithm of initial template amounts. The amounts of starting templates in mixed-infected Oncidium flowers and leaves were estimated from the standard curves. As little as 10(4) copies or 5 fg each of CymMVand ORSV could be detected simultaneously with either the RdRp or CP gene as the target. This system offers a sensitive, high throughput and rapid method for plant virus detection. (C) 2000 Elsevier Science B.V. All rights reserved.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 04/04/20 alle ore 01:24:12