Why deplete host (human or animal) DNA before PCR-based assays for bacteria and fungi?
Why is MolYsis useful?
- A major problem of the direct
PCR-based detection of bacteria and fungi from clinical specimens is that host
DNA can exceed microbial target sequences by several thousand-fold.
- Host DNA contains unspecific binding sites e.g., 16S primers leading to unspecific binding which limits the accessibility of primers to the specific sites on the 16S rRNA gene and may lead to false-positive signals.
- The depletion of host DNA increases the
sensitivity and specificity of the detection of bacterial and fungal DNA from
direct samples.
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What is the sensitivity of detection of bacteria and fungi in blood?
Sensitivity of detection / Bacteria & Fungi Spiking experiments showed that bacteria and fungi can be detected at low titers in whole blood using real-time PCR and subsequent Sanger Sequencing. For instance, with 5 ml whole blood using MolYsis™ ...How is human DNA depleted during sample processing?
Human DNA depletion Samples are treated with a buffer lysing the human/animal cells, while bacterial and fungal cells stay intact. The released human/animal DNA is enzymatically degraded along with all other free DNA and bacterial and fungal cells ...Which samples can be used for Micro-Dx™?
Micro-Dx & Clinical sample For Micro-Dx™ the following clinical samples are applicable: Body fluids: e.g., whole blood (EDTA- or citrate-stabilized), pleural fluid, cerebrospinal fluid, ascites aspirates, broncho-alveolar lavage, blood cultures, ...What benefit has human DNA depletion before a PCR assay of bacteria and fungi?
A high proportion of human DNA can lead to non-specific primer binding to human DNA, which can result in false-positive PCR signals. In addition, an excess of human DNA limits the accessibility of primers to specific sites on the 16S/18S rRNA target ...How is the human DNA depleted?
MolYsis Technology for human DNA depletion Samples are treated with a buffer lysing the human cells, while bacterial and fungal cells stay intact. The released human DNA and other free DNA are enzymatically degraded. Afterward, the microbial cells ...