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An alternative molecular approach for rapid and specific detection of clinically relevant bacteria causing prosthetic joint infections with bacteriophage K

机译:一种替代分子方法,用于临床相关细菌的快速和特异性检测,导致噬菌体k的假体关节感染k

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摘要

Prosthetic joint infections (PJI) represent the most serious cause of prosthetic joint loosening, with high impact on patient life and health economics. Although not entirely reliable, the cultivation of intraoperative prosthetic tissue or synovial fluid remains the gold standard for determining the cause of PJI. Therefore, molecular methods are increasingly being introduced. The aim of this study was to optimize and assess an alternative molecular approach with the use of bacteriophage K for more rapid and specific detection of staphylococci in sonicate fluid (SF) of PE The best results with the method were obtained after 180 min of sample incubation with 10' PFU/mL of bacteriophage K. DNA isolation prior to qPCR analysis was confirmed unnecessary, while chloroform addition to samples after incubation with bacteriophage K improved bacterial detection by 100x. The method had a limit of detection of 6.8x10(2) CFU/mL and was found suitable for the detection of staphylococci in SF of removed prosthetic joints, giving results comparable to standard microbiological methods in just four hours. The optimized method was found fit for the purpose, offering potential advantages over the use of molecular detection methods to detect bacterial DNA.
机译:假肢关节感染(PJI)代表了假体关节松动的最严重原因,对患者生命和卫生经济学影响很高。虽然不完全可靠,但术中假体组织或滑液培养仍然是确定PJI原因的金标准。因此,越来越多地引入分子方法。本研究的目的是优化和评估使用噬菌体K的替代分子方法,用于在PE的超声液(SF)中的超速且特异性检测的葡萄球菌(SF)中的最佳效果,在样品孵育180分钟后获得了该方法的最佳效果用10'PFU / ml噬菌体K.在QPCR分析之前的DNA分离被证实,而在孵育后与噬菌体K孵育后对样品进行改善1×100x的氯仿。该方法的检测限为6.8x10(2)CFU / mL,发现适用于检测除去假肢关节的SF中的葡萄球菌,可与标准微生物方法在短短4小时内进行结果。找到了优化的方法,旨在为目的提供对使用分子检测方法来检测细菌DNA的潜在优势。

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