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.
Article
de Zh
| WPRIM
| ID: wpr-334827
Bibliothèque responsable:
WPRO
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.
Texte intégral:
1
Indice:
WPRIM
Sujet Principal:
Virologie
/
Oiseaux
/
Reproductibilité des résultats
/
Sensibilité et spécificité
/
RT-PCR
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Techniques d'amplification d'acides nucléiques
/
Diagnostic
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Sous-type H5N1 du virus de la grippe A
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Grippe chez les oiseaux
/
Génétique
Type d'étude:
Diagnostic_studies
Limites du sujet:
Animals
langue:
Zh
Texte intégral:
Chinese Journal of Virology
Année:
2008
Type:
Article