|
Molecular Microbiology News Vol. 6/2019 |
||
|
Hot Topic: Molecular Characterization of Oral Bacteria in Heart Valves and Dentine Samples |
||
|
New Publications: 1. Bacterial DNA detected on pathologically changed heart valves using 16S rRNA gene amplification (Chalupova et al. 2018) 2. Molecular characterization of intact cell-derived and cell-free bacterial DNA from carious dentine samples (Schmidt et al. 2019) 3. Applications of MolYsis™ host DNA removal in culture-independent molecular analysis of bacteria and fungi (Linow 2018) Product Highlights: · MolYsis™ Technology Microbial DNA enrichment and isolation from fluid and tissue specimens, removal of non-target host DNA · DNA-free PCR Reagents DNA-free 16S and 18S PCR reagents and assays with high amplification activity and accuracy |
||
|
New Publications |
||
|
1. Bacterial DNA detected on pathologically changed heart valves using 16S rRNA gene amplification (Chalupova et al. 2018) The aim of this study was to detect oral bacteria on pathologically changed heart valves not caused by infective endocarditis. During the study period, 41 samples of heart valves removed during heart valve replacement surgery were processed. Aliquots were cultivated aerobically and anaerobically but no bacteria could be detected by conventional culture. Bacterial DNA was extracted directly from heart valves using Ultra-Deep Microbiome Prep kit followed by a 16S rRNA gene PCR amplification using Mastermix 16S Complete kit. Positive PCR products were sequenced and the sequences analyzed using a BLAST database. Bacterial DNA was detected in 21 samples whereof nine samples contained multiple bacteria. The results showed significant appearance of bacteria on pathologically changed heart valves in patients with no symptoms of infective endocarditis. |
||
|
2. Molecular characterization of intact cell-derived and cell-free bacterial DNA from carious dentine samples (Schmidt et al. 2019) In this study, carious dentine samples were collected in the course of a selective caries excavation at two different deep dentine caries lesions in three patients. The microbial community before and after therapy was analyzed by conventional culture compared to bacterial DNA analyses using 16S rRNA gene PCR and terminal restriction fragment length polymorphism (T-RFLP) for fingerprinting community changes in order to receive information about the quantitative reduction of bacterial DNA as well as microbial shifts and differences in diversity correlating with the treatment interventions. For differentiation between cell-derived and cell-free DNA two protocols based on the MolYsis™ Complete5 kit were used and bacterial DNA amplified with Mastermix 16S Complete kit. The results of this study reveal that differentiation and quantification of DNA from intact bacteria and cell-free DNA is reasonable when considering carious dentine samples, especially after intervention. |
||
|
3. Applications of MolYsis™ host DNA removal in culture-independent molecular analysis of bacteria and fungi (Linow 2018) In samples from infected humans and animals host DNA is present in vast excess to microbial DNA. Unspecific primer binding to host DNA decreases the sensitivity and specificity of molecular analysis of the microbial DNA. The MolYsis™ technology provides a solution to this problem by removing the host DNA before extraction of microbial DNA. This application note summarizes clinical and other applications of MolYsis™ sample pre-treatment in conjunction with a variety of analytical methods, including next generation sequencing, amplicon cloning, real-time PCR, Sanger sequencing and DNA array. |
||
|
Product Highlights |
||
|
|
MolYsis™ Technology Microbial DNA enrichment and isolation from fluid, swab and tissue specimens, removal of non-target host DNA Molzym’s MolYsis™-based manual and automated kits are the perfect tool for molecular microbiology and microbiome studies allowing significantly deeper sequencing compared with conventional DNA isolation systems. Product features: · Removal 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, swabs and tissues · Contamination-free DNA isolation reagents, buffers and columns |
|
|
|
Contamination-free PCR Reagents DNA-free 16S and 18S PCR reagents and assays with high amplification activity and accuracy A series of 16S/18S PCR assays, master mixes and Taq DNA polymerases are offered for contamination-free detection of microorganisms, e.g. from clinical samples. Product features: · Guaranteed microbial DNA-free · High amplification activity over 40 cycles · Ultra-sensitive detection of bacteria and fungi · Suitable for PCR and Real-Time PCR · Purification service for custom assays |
|
|
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. |
|
|
|
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, Michael Lorenz, Ph. D. (Editor in Chief) |
||
|
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. |
||
