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@ARTICLE{Fawzy:180441,
      author       = {A. Fawzy and A.-S. Giel and L. Fenske and A. Bach and C.
                      Herden and K. Engel and E. Heuser and M. Boelhauve and R. G.
                      Ulrich and K. Vogel$^*$ and K. Schmidt$^*$ and T. Eisenberg},
      title        = {{D}evelopment and validation of a triplex real-time q{PCR}
                      for sensitive detection and quantification of major rat bite
                      fever pathogen {S}treptobacillus moniliformis.},
      journal      = {Journal of microbiological methods},
      volume       = {199},
      issn         = {0167-7012},
      address      = {New York, NY},
      publisher    = {Elsevier},
      reportid     = {DKFZ-2022-01336},
      pages        = {106525},
      year         = {2022},
      note         = {Volume 199, August 2022, 106525},
      abstract     = {Streptobacillus (S.) moniliformis is the most important
                      pathogen causing rat bite fever (RBF) worldwide. This
                      zoonotic pathogen is understudied mainly due to difficulties
                      in culturing S. moniliformis as a fastidious microorganism.
                      Therefore, advances in molecular detection techniques are
                      highly needed, especially with regard to the widespread
                      availability of real-time quantitative (q) PCR in
                      laboratories. In this study, we aimed to develop a qPCR for
                      the identification of Streptobacillus species and
                      quantification of S. moniliformis in clinical samples,
                      especially those derived from tissue samples of animal
                      origin. We optimized a previously described PCR protocol in
                      order to develop a qPCR, which can detect different
                      Streptobacillus species with high specificity and is
                      simultaneously able to quantitate S. moniliformis in
                      different clinical matrices. The qPCR exhibited a limit of
                      detection (LOD) of 21 copies/reaction representing ~4-5
                      streptobacilli, while the limit of quantification (LOQ) was
                      2.1 × 103 copies/reaction. It was also more sensitive than
                      conventional PCR by two orders of magnitude and proved to
                      have a substantial agreement (Kappa 0.74) compared to it
                      with a superior detection rate in 374 samples from wild
                      rats, laboratory rats and animals from holdings of
                      wild-trapped rats. To conclude, the qPCR described in this
                      study is an important molecular tool that is able to
                      quantify S. moniliformis in tissue samples of animal origin.
                      It represents a suitable tool for future establishment and
                      evaluation of other molecular assays that are highly needed
                      for a better understanding of epidemiology and
                      pathophysiology of RBF. In experimental studies, it will
                      also be useful for titration purposes since the
                      quantification of the organism using classical plate
                      counting technique is problematic and inaccurate.},
      keywords     = {Quantification (Other) / Rat bite fever (Other) / Real-time
                      PCR (Other) / Streptobacillus moniliformis (Other)},
      cin          = {W440},
      ddc          = {570},
      cid          = {I:(DE-He78)W440-20160331},
      pnm          = {316 - Infektionen, Entzündung und Krebs (POF4-316)},
      pid          = {G:(DE-HGF)POF4-316},
      typ          = {PUB:(DE-HGF)16},
      pubmed       = {pmid:35738493},
      doi          = {10.1016/j.mimet.2022.106525},
      url          = {https://inrepo02.dkfz.de/record/180441},
}