Tural Bilsen, Ertaş Erdal, Uzuner Nurullah, Bektaş Buşra, Tural Emre, Çavdar Mehmet, Temiz Hakan, Özbek Erdal, Tural Servet
Department of Nanotechnology, Institute of Science, Dicle University, 21280, Diyarbakir, Turkey.
Department of Chemistry, Institute of Science, Dicle University, 21280, Diyarbakir, Turkey.
World J Microbiol Biotechnol. 2025 Jul 1;41(7):240. doi: 10.1007/s11274-025-04463-y.
Rapid and accurate detection of microorganisms in sterile body fluids, particularly cerebrospinal fluid (CSF), is crucial for effective diagnosis and treatment. Conventional methods, such as centrifugation, may result in low microbial recovery and false negatives, limiting diagnostic accuracy. An alternative, efficient, and accessible microbial concentration method is needed. This study evaluates a nanoparticle-based microbial concentration method to enhance pathogen recovery from CSF. The method was optimized for interaction time (1 min) and nanoparticle dosage (0.01 g/mL) using standard microbial strains, including Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa, and Candida albicans. Its clinical performance was assessed using 800 CSF samples, and microbial identification was confirmed via MALDI-TOF MS. The method significantly improved microbial recovery across 10-10 CFU/mL concentrations, achieving a detection limit as low as 2 CFU/mL. Clinical validation demonstrated 100% sensitivity and specificity, detecting 15 additional true-positive cases missed by centrifugation. While centrifugation fails to detect bacteria below 10 CFU/mL, our method reliably detects even at 10 CFU/mL, demonstrating superior microbial enrichment, especially in low-biomass samples. This method enhances diagnostic accuracy by reducing false negatives and expediting pathogen detection. Its resource-conscious, low-cost and equipment-free nature makes it particularly beneficial for resource-limited laboratories, offering a scalable alternative for microbial concentration in CSF diagnostics.
快速准确地检测无菌体液中的微生物,尤其是脑脊液(CSF),对于有效诊断和治疗至关重要。传统方法,如离心法,可能导致微生物回收率低和假阴性,限制了诊断准确性。因此需要一种替代的、高效且易于使用的微生物浓缩方法。本研究评估了一种基于纳米颗粒的微生物浓缩方法,以提高脑脊液中病原体的回收率。该方法使用包括金黄色葡萄球菌、粪肠球菌、大肠杆菌、铜绿假单胞菌和白色念珠菌在内的标准微生物菌株,针对相互作用时间(1分钟)和纳米颗粒剂量(0.01 g/mL)进行了优化。使用800份脑脊液样本评估其临床性能,并通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)确认微生物鉴定。该方法显著提高了10-10 CFU/mL浓度范围内的微生物回收率,检测限低至2 CFU/mL。临床验证显示敏感性和特异性均为100%,检测出15例离心法漏检的额外真阳性病例。虽然离心法无法检测到低于10 CFU/mL的细菌,但我们的方法即使在10 CFU/mL时也能可靠检测,显示出卓越的微生物富集能力,尤其是在低生物量样本中。该方法通过减少假阴性和加快病原体检测提高了诊断准确性。其资源节约、低成本且无需设备的特性使其对资源有限的实验室特别有益,为脑脊液诊断中的微生物浓缩提供了一种可扩展的替代方法。