光动力灭活在尿路感染病原菌中的应用

N. Ignatova, V. Elagin, I. Budruev, A. Antonyan, O. Streltsova, V. Kamensky
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摘要

光动力失活(PDI)是一种替代抗生素治疗的微生物杀灭方法,可用于膀胱腔的碎石和卫生。目的:光动力失活(PDI)对尿路病原微生物的参数选择及应用。材料与方法。在这项研究中,我们使用从患者尿液样本中分离的细菌菌株。微生物鉴定采用分化培养基和生化板。在纯培养物和天然尿液中研究了尿路病原微生物对PDI的敏感性。工作中使用光敏剂“photodiazine”(50µg/ml),并使用Triton X-100(5%体积)来增加革兰氏阴性微生物细胞壁的通透性。用波长为662 nm的医用激光装置“Latus-K”照射样品。为了评估PDI的有效性,计算了微生物菌落形成单位(CFU)的对数递减值。采用Statistica 10.0和Mann-Whitney标准进行统计学分析。从36份本地尿液中分离出尿路病原菌50株,属18种。其中以金黄色葡萄球菌、大肠杆菌、铜绿假单胞菌、肺炎克雷伯菌最为常见。革兰氏阳性菌CFU的对数下降值为5 ~ 6,对应于样品中99.999 ~ 99.9999%的细菌细胞失活。对于革兰氏阴性菌株,该值略低,范围为4 ~ 5.5,但对应于CFU细菌的灭活率为99.99 ~ 99.999%。加入Triton X-100后,大肠杆菌的效率从46%提高到99.99%,P. mirabilis的效率从99%提高到99.99%,肺炎克雷伯菌的效率从16%提高到94%,铜绿假单胞菌的效率从97%提高到99.999%。结论:光动力灭活可作为一种替代抗生素治疗方法,对泌尿系病原微生物具有杀灭作用。
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Application of photodynamic inactivation against pathogens of urinary tract infections
Photodynamic inactivation (PDI) is an alternative to antibiotic therapy method for biocidal action against microorganisms, which can be used for lithotripsy and sanitation of the bladder cavities. Objective. Selection of parameters and application PDI against uropathogenic microorganisms. Materials and Methods. In this study we used bacterial strains isolated from urine samples of patients. Differentiation media and biochemical plates were used for identification of microorganisms. The sensitivity of uropathogenic microorganisms to PDI was studied on pure cultures and in native urine. The photosensitizer “Photoditazine” (50 µg/ml) was used in the work, as well as Triton X-100 (5 % vol.) was applying to increase the permeability of the cell wall of gram-negative microorganisms. The samples were irradiated by a medical laser device “Latus-K” with a wavelength of 662 nm. To assess the effectiveness of PDI, the values of the logarithmic decrease of colony-forming unit (CFU) of the microorganisms were calculated. Statistical analysis was made by Statistica 10.0 and Mann-Whitney criterion. Results. 50 strains of uropathogens belonging to 18 species were isolated from 36 samples of native urine. Among them, the most common were S. aureus, E. coli, P. aeruginosa, K. pneumoniae. The value of logarithmic decrease in CFU for gram-positive bacteria ranged from 5 to 6, which corresponds to inactivation 99.999-99.9999% of bacterial cells in a sample. For gram-negative strains, this value was slightly lower and ranged from 4 to 5.5, which, nevertheless, corresponds to inactivation 99.99-99.999% of CFU bacteria. The addition of Triton X-100 increase the efficiency from 46% to 99.99% for E. coli, from 99% to 99.99% for P. mirabilis, from 16% to 94% for K. pneumoniae and from 97% to 99.999% for P. aeruginosa. It should be noted that the PDI was affect microorganisms both in isolated pure cultures and in native urine. Conclusions. Photodynamic inactivation may be considered as an alternative to antibiotic therapy method of biocidal action against uropathogenic microorganisms.
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