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Molecular Microbiology News Vol. 1/2020 |
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Hot Topic: MolYsis™ Host DNA Depletion for Deeper Microbiome and Metagenome Analysis |
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New Publications: 1. Rapid detection of difficult-to-culture bacterial pathogens using real-time nanopore sequencing (Gündoğdu et al. 2019) 2. Application of metagenomic shotgun sequencing to detect vector-borne pathogens in clinical blood samples (Vijayvargiya et al. 2019) 3. MolYsis™ host DNA depletion for deeper microbiome and metagenome analysis [updated version] (Wollens 2020) Product Highlights: · MolYsis™ Basic5 & MolYsis™ Complete5 Depletion of non-target host DNA for microbial DNA enrichment from fluid specimens · Ultra-Deep Microbiome Prep Microbial DNA enrichment and isolation from fluid and tissue specimens, depletion of non-target host DNA |
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New Publications |
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1. Rapid detection of difficult-to-culture bacterial pathogens using real-time nanopore sequencing (Gündoğdu et al. 2019) In this report, two different cases of suspected peritoneal tuberculosis and brucellosis, without and with delayed confirmation by conventional microbiology, are described. Nanopore-based shotgun metagenomic sequencing was applied to rapidly detect the infectious agents from peritoneal fluid and blood. Samples were prepared with MolYsis™ Complete5 including human DNA depletion and isolation of microbial DNA followed by sequencing of the DNA on a MinION. The earliest runtime meeting the confident detection threshold was only after 20 minutes for Mycobacterium tuberculosis and after 30 minutes for Brucella melitensis. A high confidence level of detection at the species level was achieved within six hours. The real-time nanopore sequencing technology allowed the authors to achieve the results within six hours with low capital costs and minimal equipment need. In comparison to conventional microbial and molecular diagnostic methods, this shortened the turnaround time and decreased the costs significantly. |
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2. Application of metagenomic shotgun sequencing to detect vector-borne pathogens in clinical blood samples (Vijayvargiya et al. 2019) Metagenomic shotgun sequencing (MGS) as non-hypothesis driven diagnostic test was evaluated in this publication by Vijayvargiya and his colleagues. Eight EDTA blood samples were analysed conventionally by microscopy and laboratory-developed PCR assays. For MGS, MolYsis™ Complete5 was used for the depletion of host DNA and isolation of microbial DNA, followed by amplification with REPLI-g Single Cell Kit and sequencing on a HiSeq® 2500 in rapid run mode. Four different analytical tools have been compared and at least one tool was able to detect four out of six organisms at the genus level and five of eight at the species level of vector-borne human pathogens including protozoa and one helminth. The filarial worm Mansonella perstans was missed by all tools; however, genetic material of M. perstans could be detected. The authors highlight that instead of single gene amplification approaches based on clinical suspicions or hypothesis, metagenomic shotgun sequencing allows to detect bacteria, viruses, fungi, protozoa and helminths in a single test. |
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3. MolYsis™ host DNA depletion for deeper microbiome and metagenome analysis [updated version] (Wollens 2020) In many samples from infected humans and animals host DNA greatly outnumbers microbial DNA. Unspecific primer binding to host DNA decreases the power of resolution of microbiomes and metagenomes. The MolYsis™ technology provides a solution to this problem by depleting the host DNA before isolation of microbial DNA. The technology is available for fluid and tissue samples, manually and automated. In this application note, Wollens summarizes a variety of studies including the MolYsis™ technology in combination with major NGS systems as a means of marked increase of microbial reads and hence deeper analysis of microbiomes. |
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Product Highlights |
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MolYsis™ Basic5 & MolYsis™ Complete5 Microbial DNA enrichment from fluids, depletion of non-target host DNA 2 optimal solutions are available for i) integration into validated in-house DNA isolation systems with the MolYsis™ Basic5 and ii) the complete isolation solution with the MolYsis™ Complete5 including contamination-free columns, enzymes and buffers for molecular microbiology and microbiome studies. Product features: · Validated for a variety of body fluids · Depletion of non-target human/animal DNA (MolYsis™ technology) · Isolation of enriched microbial DNA (MolYsis™ Complete5) · Broad-range lysis of bacteria and fungi · Small and large volumes: ≤1 ml and 5 ml liquid samples · Automation available |
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Ultra-Deep Microbiome Prep Microbial DNA enrichment and isolation from fluid and tissue specimens, depletion of non-target host DNA Molzym’s Ultra-Deep Microbiome Prep kit allows significantly deeper sequencing in microbiome studies compared with conventional DNA isolation systems. Product features: · Validated for a variety of body fluids and tissue specimens · Depletion of non-target human/animal DNA (MolYsis™ technology) · Isolation of enriched microbial DNA · Broad-range lysis of bacteria and fungi · Small and large volumes: 0.2 to 10 ml liquid samples and 0.25cm³ tissues · Contamination-free DNA isolation reagents, buffers and spin columns · Automation available |
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Follow Molzym’s Blog! Stay informed on topics covering molecular diagnostics, pathogen enrichment and DNA isolation from clinical samples, automation and next generation sequencing. Get practical tips and insights into our solutions. |
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Dear reader, Welcome to this edition of Molecular Microbiology News. With this newsletter we want to keep you updated on the latest developments in molecular detection of microbial infections. Best regards, Your Molzym team |
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This information is brought to you by Molzym GmbH & Co. KG, Mary-Astell-Str.10, D-28359 Bremen, Germany // If this information is not of interest to you, please disregard and we apologize for any inconvenience this may have caused. If you want to unsubscribe from this newsletter please click unsubscribe. |
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