![]() Finally, the JEV RT-LAMP assay was simpler and less time consuming than conventional RT-PCR or real-time RT-PCR, since the amplification step could be completed in a single tube within 50 min at 63☌. The results of virus isolation and identification of swine blood samples and mosquito samples were fully consistent with RT-LAMP. Assay specificity was high, showing no cross-reactivity to other flaviviruses. Assay sensitivity was similar to real-time RT-PCR, but was 10-fold higher than conventional RT-PCR. RT-LAMP had detection limits of 2.57 and 2.34 copies/μL for JEV I and III, respectively. Fifty-six swine blood samples and 20,000 mosquitoes were used to evaluate the method, compared to conventional RT-polymerase chain reaction (PCR) and real-time RT-PCR. In this study, a rapid JEV detection method for swine and mosquitoes was developed based on reverse transcription loop-mediated isothermal amplification (RT-LAMP) targeting the nucleocapsid (E) genes of JEV genotype I (lineage K94PO5), and genotype III (lineage SA14-14-2). Porcine infection results in fatal encephalitis, abortion, and stillbirth in pregnant sows, and hypospermia in boars. One of the natural hosts of the mosquito-borne JE virus (JEV) is domestic pigs, which act as amplifier hosts. Japanese encephalitis (JE) can infect many agriculturally important animals and humans, and has a high incidence in Asia.
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