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Titolo:
A comprehensive systematic approach to identification of influenza A virusgenotype using RT-PCR and RFLP
Autore:
Offringa, DP; Tyson-Medlock, V; Ye, ZP; Levandowski, RA;
Indirizzi:
US FDA, Ctr Biol Evaluat & Res, Div Viral Prod, Rockville, MD 20852 USA USFDA Rockville MD USA 20852 es, Div Viral Prod, Rockville, MD 20852 USA
Titolo Testata:
JOURNAL OF VIROLOGICAL METHODS
fascicolo: 1, volume: 88, anno: 2000,
pagine: 15 - 24
SICI:
0166-0934(200007)88:1<15:ACSATI>2.0.ZU;2-C
Fonte:
ISI
Lingua:
ENG
Soggetto:
NUCLEOTIDE-SEQUENCES; RECOMBINANTS; VACCINES; SEGMENTS;
Keywords:
influenza virus; influenza vaccine reassortants; RT-PCR; RFLP; genotype 1;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
19
Recensione:
Indirizzi per estratti:
Indirizzo: Offringa, DP US FDA, Ctr Biol Evaluat & Res, Div Viral Prod, 1401 Rockville Pike HFM 463, Rockville, MD 20852 USA US FDA 1401 Rockville Pike HFM 463 Rockville MD USA 20852 USA
Citazione:
D.P. Offringa et al., "A comprehensive systematic approach to identification of influenza A virusgenotype using RT-PCR and RFLP", J VIROL MET, 88(1), 2000, pp. 15-24

Abstract

Amplification of influenza A virus gene segments by reverse transcription-polymerase chain reaction (RT-PCR) can be combined with enzymatic digestionto reveal unique restriction fragment length polymorphisms specific for H1N1 and H3N2 subtype viruses. We have used the method to provide a rapid, specific and reproducible identification of the genotype of high-growth influenza reassortants derived from A/Puerto Rico/8/34 (PR8). Digestion of the gene segments amplified from wild-type viruses, PR8 and reassortants at sites unique to either the wild-type strain or to PR8 provided positive, unambiguous identification of the origin of each of the internal genes, and distinguished the internal genes of both H1N1 and H3N2 strains from those of PR8. This method has also permitted us to quickly confirm that reasserting hasoccurred and to optimize the selection of reassortant clones with maximum number of PR8 internal genes. Since the method can detect 1-10% of a secondstrain in a mixed population, the method can also be used to detect samples containing more than one viral subtype and to assess the purity of influenza viruses used for manufacturing vaccines. (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 30/03/20 alle ore 13:34:58