How Many Microorganisms Are Present? Quantitative Reverse Transcription PCR (qRT-PCR)

Price, Andy; Álvarez, Laura Acuña; Whitby, Corinne; Larsen, Jan

doi:10.1007/978-90-481-9252-6_9

Cited by 1 publication

(1 citation statement)

References 12 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Reverse transcription qPCR (qRT-PCR), which targets mRNA in actively metabolizing cells, can also be used for absolute quantification with a higher sensitivity compared to regular qPCR. Here, mRNA is converted to cDNA via reverse transcription and subsequently quantified using qPCR [ 46 ]. qRT-PCR has been used for bacterial detection and quantification when it is necessary to determine the number of active bacteria, such as in clinical settings, food industry, and activated sludge (wastewater remediation) [ 47 , 48 , 49 , 50 ].…”

Section: Brief Description Of Advanced Absolute Quantification Methodsmentioning

confidence: 99%

Current Applications of Absolute Bacterial Quantification in Microbiome Studies and Decision-Making Regarding Different Biological Questions

et al. 2021

View full text Add to dashboard Cite

High throughput sequencing has emerged as one of the most important techniques for characterizing microbial dynamics and revealing bacteria and host interactions. However, data interpretation using this technique is mainly based on relative abundance and ignores total bacteria load. In certain cases, absolute abundance is more important than compositional relative data, and interpretation of microbiota data based solely on relative abundance can be misleading. The available approaches for absolute quantification are highly diverse and challenging, especially for quantification in differing biological situations, such as distinguishing between live and dead cells, quantification of specific taxa, enumeration of low biomass samples, large sample size feasibility, and the detection of various other cellular features. In this review, we first illustrate the importance of integrating absolute abundance into microbiome data interpretation. Second, we briefly discuss the most widely used cell-based and molecular-based bacterial load quantification methods, including fluorescence spectroscopy, flow cytometry, 16S qPCR, 16S qRT-PCR, ddPCR, and reference spike-in. Last, we present a specific decision-making scheme for absolute quantification methods based on different biological questions and some of the latest quantitative methods and procedure modifications.

show abstract

Section: Brief Description Of Advanced Absolute Quantification Methodsmentioning

confidence: 99%

Current Applications of Absolute Bacterial Quantification in Microbiome Studies and Decision-Making Regarding Different Biological Questions

et al. 2021

View full text Add to dashboard Cite

show abstract

How Many Microorganisms Are Present? Quantitative Reverse Transcription PCR (qRT-PCR)

Cited by 1 publication

References 12 publications

Current Applications of Absolute Bacterial Quantification in Microbiome Studies and Decision-Making Regarding Different Biological Questions

Current Applications of Absolute Bacterial Quantification in Microbiome Studies and Decision-Making Regarding Different Biological Questions

Contact Info

Product

Resources

About