As the 16S profiles in the intestine seem extremely various for intracellular and extracellular DNA [33], qPCR is predicted to mirror the NGS-specific Group composition both quantitatively and qualitatively more carefully than stream cytometry.
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qPCR is comparatively straightforward to perform as compared to movement cytometry that needs significant abilities for reproducible success. Actually, flow cytometric enumeration of microbial cells was to begin with restricted to pure cultures [31] and however continues to be difficult when performed in complex matrices [32].
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The use of relative abundance facts from next generation sequencing (NGS) may lead to misinterpretations of microbial Group buildings, as the increase of one taxon leads to the concurrent lower of the opposite(s) in compositional details. Though distinct DNA- and cell-dependent solutions along with statistical techniques are already developed to beat the compositionality dilemma, and also the Organic relevance of absolute bacterial abundances has long been shown, the human microbiome exploration hasn't nonetheless adopted these solutions, most likely on account of feasibility problems.
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