Specific detection of RT activity in culture supernantants of retrovirus-producing cells, using synthetic DNA as competitor in polymerase enhanced reverse transcriptase assay.
J Virol Methods
; 94(1-2): 187-93, 2001 May.
Article
em En
| MEDLINE
| ID: mdl-11337053
ABSTRACT
The polymerase enhanced reverse transcriptase (PERT) assay is a highly sensitive assay for the detection of reverse transcriptase (RT) activity in culture supernatants of retrovirus-producing cells. However, some cellular DNA-dependent DNA polymerases exhibit RT-like activities in this assay. A synthetic DNA competitor which suppresses the RT-like activities of cellular DNA-dependent DNA polymerases was used in a modified PERT assay technique for specific detection of RT activity in culture supernatants of retrovirus-producing cells. We determined the optimum condition of the assay and evaluated its specificity. This improved PERT assay is easy to perform and is able to detect minute amounts of purified RT, as well as RT in crude cell lysates and concentrated culture supernatants.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Polidesoxirribonucleotídeos
/
Retroviridae
/
Reação em Cadeia da Polimerase
/
DNA Polimerase Dirigida por RNA
Tipo de estudo:
Diagnostic_studies
Limite:
Animals
Idioma:
En
Revista:
J Virol Methods
Ano de publicação:
2001
Tipo de documento:
Article
País de afiliação:
França