I am running qRT-PCR for detecting Zika ARN on serum and urine samples and we cannot keep up with the quantity of samples we get to analyse. There is many people in our institute that say we cannot do sample pooling with human samples, eventhough the prevalence is really low. Is someone using sample pooling for any viral ARN or DNA detection technique and have evidence that it works and it is possible to maintain sensitivity?
Look at Roche COBAS MPX, minipools are in use now for many years (since 2000) for detecting blood borne viruses in blood donation.
It might be possible if you first isolate the RNA or DNA from each sample. Eluate each sample (I would recommend up to 5 samples) with 20 or 30 µl and absorp it again on the columbs with all washing steps. Then eluate it into 50 µl. That can work. We did this with DNA and it was working.
It is possible, but I am not sure if it is advisable in your instance. Pooling 5ul of each RNA purification elution into a pool of 5-10 individuals works well for broad surveillance screens but should never be adopted for patient diagnostics. As it sounds like you are performing frontline diagnostics I would encourage you to perform the most sensitive and accurate method which is individual testing. Pooling 10 individuals will shift the Ct of any positive by 3.32 cycles - given the late Cts (in serum especially) this could have massive implications for data interpretation
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