RESUMO
A sensitive reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for rapid visual detection of Murray valley encephalitis virus (MVEV) infection. The reaction was performed in one step in a single tube at 63 °C for 60 min with the addition of the hydroxynaphthol blue (HNB) dye prior to amplification. The detection limit of the RT-LAMP assay was 100 copies per reaction based on 10-fold dilutions of in vitro transcribed RNA derived from a synthetic MVEV DNA template. No cross-reaction was observed with other encephalitis-associated viruses. The assay was further evaluated using spiked cerebrospinal fluid sample with pseudotype virus containing the NS5 gene of MVEV.
Assuntos
Sequência de Bases , Primers do DNA , Vírus da Encefalite do Vale de Murray , Genética , Limite de Detecção , Transcrição GênicaRESUMO
<p><b>OBJECTIVE</b>To develop a headspace gas chromatography method for the determination of residual organic solvents in Panax notoginseng extracts.</p><p><b>METHODS</b>The samples were injected into HP-INNOWAX capillary column by headspace sampler and analyzed with FID detector using standard addition method.</p><p><b>RESULTS</b>There was a good linearity in the experimental concentration (r=0.9932-0.9999). The rate of recovery was in the rang of 81.74%-111.2%. The numbers of theoretical plates were more than 15000 and the resolutions between the adjacent peaks were more than 2. The RSD of precision and accuracy were both less than 10 %.</p><p><b>CONCLUSION</b>The method is simple,accurate and sensitive with good reproducibility, which can be used for the determination of the residual organic solvents in Panax notoginseng extracts.</p>