Absolute Quantification of Viable Bacteria Abundances by Next Generation Sequencing

Next-generation sequencing is an important tool for taxonomical bacteria identification. Despite the undeniable advantages of the approach and recent developments in technology, which have led to its improvement and availability, it still has several limitations and shortcomings. The usual outcome of microbiota sequencing is a relative abundance of bacterial taxa. The information about bacteria viability or enumeration is missing. However, this knowledge is crucial for many applications. Moreover, the analysis based on only relative abundances may sometimes be deceptive. In the current study, we elaborated the complete workflow for absolute quantification of living bacteria based on 16S rRNA gene amplicon sequencing.