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. 2017 Jul;40(3):190-196. Epub 2017 Jul 4.
Staphylococcus spp. accounts for up to two thirds of all microorganisms causing prosthetic joint infections, with Staphylococcus aureus and Staphylococcus epidermidis being the major cause. The present study describes a diagnostic model to detect staphylococci using a specific bacteriophage and bioluminescence detection, exploring the possibility of its use on sonicate fluid of orthopaedic artificial joints. Intracellular adenosine-5'-triphosphate release by bacteriophage mediated lysis of staphylococci was assessed to determine optimal parameters for detection. With the optimized method, a limit of detection of around 103 CFU/mL was obtained after incubation with bacteriophage for 2 h. Importantly, sonicate fluid did not prevent the ability of bacteriophage to infect bacteria and all simulated infected sonicate fluid as well as 6 clinical samples with microbiologically proven staphylococcal infection were detected as positive. The total assay took approximately 4 h. Collectively, the results indicate that the developed method promises a rapid, inexpensive and specific diagnostic detection of staphylococci in sonicate fluid of infected prosthetic joints. In addition, the unlimited pool of different existing bacteriophages, with different specificity for all kind of bacteria gives the opportunity for further investigations, improvements of the current model and implementation in other medical fields for the purpose of the establishment of a rapid diagnosis.
葡萄球菌属占所有引起人工关节感染的微生物的三分之二，其中金黄色葡萄球菌和表皮葡萄球菌是主要病因。本研究描述了一种使用特定噬菌体和生物发光检测来检测葡萄球菌的诊断模型，探讨了其在骨科人工关节超声处理液中应用的可能性。评估了噬菌体介导的葡萄球菌裂解引起的细胞内三磷酸腺苷释放，以确定检测的最佳参数。采用优化方法，与噬菌体孵育2小时后，检测限约为103 CFU/mL。重要的是，超声处理液并未阻止噬菌体感染细菌的能力，所有模拟感染的超声处理液以及6份经微生物学证实为葡萄球菌感染的临床样本均检测为阳性。整个检测过程约需4小时。总体而言，结果表明所开发的方法有望对感染人工关节的超声处理液中的葡萄球菌进行快速、廉价且特异的诊断检测。此外，现有的不同噬菌体库对各种细菌具有不同的特异性，这为进一步研究、改进当前模型以及在其他医学领域实施以建立快速诊断提供了机会。
