Treviño Samuel, Ramírez-Flores Eduardo, Cortezano-Esteban Steffany, Hernández-Fragoso Hugo, Brambila Eduardo
Laboratory of Metabolomic and Chronic Degenerative Diseases, Physiology Institute, Meritorious Autonomous University of Puebla, Prol. de la 14 Sur 6301, Ciudad Universitaria, Puebla C.P. 72560, Mexico.
Center for Care and Research in Health Services, Urinalysis and Microbiology Area, Rio Nexapa 6153, Col. San Manuel, Puebla C.P. 72560, Mexico.
J Clin Med. 2024 Sep 9;13(17):5334. doi: 10.3390/jcm13175334.
Urinary tract infection is a worldwide health problem. According to the Clinical Laboratory Improvement Amendments and the European Urinalysis Guideline, urine samples should be tested within 2 h of collection. Thus, using chemical preservatives that guarantee the pre-analytical conditions is a practical tool. However, the effects of temperature and storage time as uropathogenic bacteria stressors are unclear. Gram-negative and -positive ATTC strains, , , , and , were used in this study. Strains in liquid media were stored at 4, 25, and 37 °C for 0, 2, 12, 24, and 48 h in tubes with and without preservatives. Then, reactive oxygen species (ROS) levels, viable but non-culturable bacteria (VBNC), and bacteria growth were analyzed. A high ROS level was associated with the presence of VBNC and dead bacteria with low CFU counts, but a low ROS level increased the CFU number, depending on temperature and storage time in tubes without preservatives (boric acid, sodium borate, and formate). The BD Vacutainer™ Urine Culture & Sensitivity Preservative PLUS Plastic Tubes (C&S-PP) prevent this ROS increase, maintaining the CFU number for longer. C&S-PP tubes minimize the stressor effects (temperature and time storage) on uropathogenic bacteria when stored, improving the pre-analytical conditions of cultures realized by the clinical laboratory.
尿路感染是一个全球性的健康问题。根据《临床实验室改进修正案》和《欧洲尿液分析指南》，尿液样本应在采集后2小时内进行检测。因此，使用能保证分析前条件的化学防腐剂是一种实用的手段。然而，温度和储存时间作为尿路致病性细菌应激源的影响尚不清楚。本研究使用了革兰氏阴性和阳性的美国典型培养物保藏中心（ATTC）菌株，即 、 、 、 和 。将液体培养基中的菌株在有和没有防腐剂的试管中于4、25和37°C下储存0、2、12、24和48小时。然后，分析活性氧（ROS）水平、活的但不可培养细菌（VBNC）和细菌生长情况。高ROS水平与VBNC的存在以及低菌落形成单位（CFU）计数的死细菌有关，但低ROS水平会增加CFU数量，这取决于无防腐剂试管（硼酸、硼酸钠和甲酸盐）中的温度和储存时间。BD Vacutainer™尿液培养与药敏防腐剂增强型塑料试管（C&S-PP）可防止这种ROS增加，使CFU数量在更长时间内保持稳定。当储存时，C&S-PP试管可将应激源（温度和储存时间）对尿路致病性细菌的影响降至最低，改善临床实验室进行培养的分析前条件。
