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Memelis, I.

Publications and source records attributed to Memelis, I..

2 recordsLinked to original sources

Genomic comparison of highly related pairs of E. coli and K. pneumoniae isolated from faeces and blood of the same neonatal patients hospitalized with fever in Dar es Salaam, Tanzania

Blood stream infections (BSIs) are a major cause of hospitalisation and death for children under the age of five in sub-Saharan Africa with members of the Gram-negative bacteria Enterobacterales such as Klebsiella pneumoniae and Escherichia coli among the most common causative agents. These bacteria usually colonise the human gastrointestinal (GI) tract which has been identified as a reservoir for invasive infections into extra-intestinal environments such as the urinary tract and bloodstream. In this study we used comparative genomics to compare hybrid genome assemblies of blood and faecal isolates taken from the same patients (all neonates under 19 days old) to determine if the BSI associated bacterial isolates originated in their GI tract. We show that both E. coli and K. pneumoniae likely translocated from the GI tract to the blood in multiple cases of BSI. We also highlight key virulence genes and acquired mutations that are indicative of pathogenic strains capable of BSI. These findings expand our understanding of BSI pathogenesis and could help guide targeted interventions to prevent future BSI infections in neonates.

genomics↗

A mycobacterial DNA extraction protocol designed for resource limited settings generates high quality whole genome sequencing

Mycobacteria are major global human pathogens and include Mycobacterium tuberculosis, the causative agent of tuberculosis, and M. abscessus, an emerging multidrug resistant pathogen. M. abscessus affects people with structural lung disease and those who are immunocompromised, most commonly causing pulmonary disease but also disseminated infections in the central nervous system and skin. High quality whole genome sequencing is essential to research mycobacterial epidemiology, pathogenesis and antimicrobial resistance. However, current DNA extraction protocols are time consuming, use toxic chemicals, require cold chain storage for certain reagents and can often result in poor quality, degraded DNA that directly impacts whole genome sequencing outputs. This is a particular challenge in low-income settings. Here, we report a novel optimised DNA extraction workflow for M. tuberculosis and M. abscessus that invariably generates high quality Illumina short read sequencing data. We evaluated input culture CFU and physical cell disruption times. DNA quantity was determined using a Qubit fluorometer system with DNA integrity assessed using the Agilent TapeStation platform. We showed that this protocol facilitated complete genome assemblies of M. abscessus and M. tuberculosis reference strains. There is no requirement for cold chain transport or storage of reagents, solvent extractions, or boiling to heat inactivate cultures, and the method does not require surfactant chemicals such as cetyltrimethylammonium bromide (CTAB).

microbiology↗