Hi could anyone share their practical experience about Arboviruses detection from field caught mosquitoes. I have used mosquito samples to perform metagenomic sequencing and passaged on C6/36 cells for Arboviruses detection. I have detected a considerable number of reads for JEV/GETV and the similar samples produced CPE’s on C6/36 cells and also confirmed the particular virus presence by RT-PCR from CPE’s positve C6/36 cells (Japanese encephalitis virus, GETV etc). However when I passaged the same samples on BHK cells, it don’t produce CPE’s.
Thank you in advance.
In the lab always keep mosquitoes at 4 degrees C. Homogenize the mosquitoes (pool mosquitoes in groups of 25-50 mosquitoes/ group). Centrifuge each group at 3000 rpm for 10 minutes and use the supernatant in rt-PCR to detect the virus.
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