What are the best methods for nasopharyngeal swab processing for viral q-PCR detection?
After collection of sample and cut off the swab tip in the UTM then vigorously agitate swabs in medium for 15-20 sec.Each sample was aliquoted and stored at -80°C until further analysis.
My question should I centrifuge the sample to get rid of debris before aliquot and storage it or after that? What are the best speed and time for centrifugation? then I should consider the supernatant or the pellet for RNA Extraction?
you do not need to centrifuge but if you are using -80 stored sample firstly come on room temp, then vortex for 10-15 sec and then you can use sample for RNA extraction,because in vtm no extra supplement provided by which virus can double in number.
If you get rid of the epithelial cells debris, you get rid of intracellular viruses as well (at least in part). It is good to make it in way described by Satendra Chandel. You can also use short term storage at -20.
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