Quantification of Bacterial DNA for Metagenomic Control Materials
Conference Recording Aug 25, 2014
About the Speaker
Denise obtained her PhD at University College London investigating the evolution of molecular mechanisms of drug resistance in Mycobacterium tuberculosis. Postdoctoral work has included the investigation of the relationship between bacterial fitness and genotype at UCL, molecular TB diagnostic methods using isothermal technologies and mass spectrometry and the investigation of factors which influence the emergence of bacterial resistance using next generation sequencing technologies at the London School of Hygiene & Tropical Medicine. Denise moved to LGC in 2011 working in the Molecular and Cell Biology Team on projects such as Metagenomics for Microbial Analysis, International Reference Measurement Methods for Nucleic Acid Metrology, Improved Measurement of Gene Regulation Processes and Metrology for Monitoring Infectious Disease and Antimicrobial Resistance in Microorganisms. Abstract
Advances in DNA sequencing technology enable the analysis of the genetic content of microbial populations (the metagenome) as a whole. Associated studies commonly measure the different quantities of the component organisms. Early evidence suggests however that sample processing and preparation methods can have a strong influence on the measured abundance of the respective microbes. In this study we use digital PCR to quantify the component organisms of a metagenomic control material comprising 10 common human pathogens mixed together in different proportions. We investigate how targeting different bacterial chromosomal positions effect the measurement, compare dPCR with fluorescence and absorbance based methods as well as next generation sequencing approaches. We demonstrate how dPCR may offer a simple method for providing confidence when measuring complex metagenomes.