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000180441 0247_ $$2doi$$a10.1016/j.mimet.2022.106525
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000180441 037__ $$aDKFZ-2022-01336
000180441 041__ $$aEnglish
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000180441 1001_ $$aFawzy, Ahmad$$b0
000180441 245__ $$aDevelopment and validation of a triplex real-time qPCR for sensitive detection and quantification of major rat bite fever pathogen Streptobacillus moniliformis.
000180441 260__ $$aNew York, NY$$bElsevier$$c2022
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000180441 500__ $$aVolume 199, August 2022, 106525
000180441 520__ $$aStreptobacillus (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.
000180441 536__ $$0G:(DE-HGF)POF4-316$$a316 - Infektionen, Entzündung und Krebs (POF4-316)$$cPOF4-316$$fPOF IV$$x0
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000180441 650_7 $$2Other$$aQuantification
000180441 650_7 $$2Other$$aRat bite fever
000180441 650_7 $$2Other$$aReal-time PCR
000180441 650_7 $$2Other$$aStreptobacillus moniliformis
000180441 7001_ $$aGiel, Ann-Sophie$$b1
000180441 7001_ $$aFenske, Linda$$b2
000180441 7001_ $$aBach, Alexa$$b3
000180441 7001_ $$aHerden, Christiane$$b4
000180441 7001_ $$aEngel, Katharina$$b5
000180441 7001_ $$aHeuser, Elisa$$b6
000180441 7001_ $$aBoelhauve, Marc$$b7
000180441 7001_ $$aUlrich, Rainer G$$b8
000180441 7001_ $$0P:(DE-He78)9029e7d8e1a0081844702984bb4a06be$$aVogel, Klaus$$b9$$udkfz
000180441 7001_ $$0P:(DE-He78)4e5ebf211bad5a927f9a00c8e9c1d116$$aSchmidt, Katja$$b10$$udkfz
000180441 7001_ $$aEisenberg, Tobias$$b11
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