Šuster Katja, Podgornik Aleš, Cör Andrej
Valdoltra Orthopaedic Hospital, Research department, Ankaran, Slovenia.
University of Ljubljana, Faculty of Chemistry and Chemical Technology, Ljubljana, Slovenia.
New Microbiol. 2020 Jul;43(3):107-114. Epub 2020 Jun 29.
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 PJI. The best results with the method were obtained after 180 min of sample incubation with 104 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 100×. The method had a limit of detection of 6.8×102 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.
人工关节感染(PJI)是人工关节松动最严重的原因,对患者生活和健康经济学有重大影响。尽管并非完全可靠,但术中人工关节组织或滑液培养仍是确定PJI病因的金标准。因此,分子方法正越来越多地被引入。本研究的目的是优化并评估一种使用噬菌体K的替代分子方法,以更快速、特异地检测PJI超声处理液(SF)中的葡萄球菌。用104 PFU/mL的噬菌体K对样品孵育180分钟后,该方法取得了最佳结果。qPCR分析前确认无需进行DNA分离,而在与噬菌体K孵育后向样品中添加氯仿可使细菌检测提高100倍。该方法的检测限为6.8×102 CFU/mL,适用于检测取出的人工关节SF中的葡萄球菌,在短短四小时内给出与标准微生物学方法相当的结果。优化后的方法被认为适用于此目的,与使用分子检测方法检测细菌DNA相比具有潜在优势。