Evaluation of reverse transcription loop-mediated isothermal amplification for detection of avian influenza A H5N1 virus / 病毒学报
Chinese Journal of Virology
;
(6): 178-184, 2008.
Artigo
em Chinês
| WPRIM
| ID: wpr-334827
ABSTRACT
A simple and sensitive Reverse Transcription Loop-mediated Isothermal Amplification (RT-LAMP) method was established to provide a new alternative for clinical diagnosis of Avian influenza A H5N1 virus. The method employed a set of six specially designed primers that recognized eight distinct sequences of the target for amplification of nucleic acid under isothermal conditions. In current study, fifty-one experimentally infected animal specimens and viral cultures that had been tested were analyzed by RT-LAMP for NA gene and HA gene, respectively. The amplification process of LAMP was monitored in real-time by the addition of SYBR Green dye. Meanwhile, the result showed high correlation between nested PCR and RT-LAMP. The specificity of the RT-LAMP assay was confirmed by restriction enzyme digestion analysis and sequencing of the amplified product. When the sensitivity of this assay was tested by serial 10-fold dilutions of RNA molecules from specimens, it was found that the RT-LAMP method achieved theoretically a sensitivity of 10 RNA molecules. Thus, we concluded that the RT-LAMP assay has potential usefulness for rapid detection of the Avian influenza A H5N1 virus.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Virologia
/
Aves
/
Reprodutibilidade dos Testes
/
Sensibilidade e Especificidade
/
Reação em Cadeia da Polimerase Via Transcriptase Reversa
/
Técnicas de Amplificação de Ácido Nucleico
/
Diagnóstico
/
Virus da Influenza A Subtipo H5N1
/
Influenza Aviária
/
Genética
Tipo de estudo:
Estudo diagnóstico
Limite:
Animais
Idioma:
Chinês
Revista:
Chinese Journal of Virology
Ano de publicação:
2008
Tipo de documento:
Artigo
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