The two important observations of researchers on our study are regarding the variability and non-comparability of results at different sites due to non-uniformity in the kits and protocols used for RNA extraction and RT-PCR.
We appreciate the observations of the researchers. It is always ideal to have the same kits, batches and study protocols to be used to ensure that the results are comparable. However, our aim was to understand the feasibility of pooled sample testing in real-life settings. We performed the pooling experiment for five and 10-samples in the field in real-life situations. The confounding factors such as quality of sample collected, level of training of the field worker, type of viral transport media and swabs used, transportation and storage of samples were not modified to make it ideal. Similarly, as in real-life situations, different kits and PCR cycles for amplification were used. The results thus obtained give us an understanding of the level to which pooling would work in the field in actual settings.