Théron Marie-Laure, Piane Laetitia, Lucarelli Laetitia, Henrion Rémi, Layssol-Lamour Catherine, Palanché Florence, Concordet Didier, Braun Jean-Pierre D, Trumel Catherine, Lavoué Rachel
Am J Vet Res. 2017 Aug;78(8):990-999. doi: 10.2460/ajvr.78.8.990.
OBJECTIVE To investigate effects of storage conditions on the canine urine protein-to-creatinine ratio (UPC) and on SDS-agarose gel electrophoresis (AGE) of urinary proteins. SAMPLE Urine specimens from 20 proteinuric (UPC > 0.5) and 20 nonproteinuric (UPC ≤ 0.2) dogs. PROCEDURES UPC and SDS-AGE were performed on urine specimens stored at room temperature (20°C) and 4°C for up to 5 days and at -20° and -80°C for up to 360 days; some specimens were subjected to 3 freeze-thaw cycles. Results were compared with those obtained for fresh urine specimens. RESULTS UPC was not affected by storage at room temperature or by freezing. A decrease in UPC was observed for specimens from nonproteinuric dogs after 5 days at 4°C (10%) and from both groups after 90 days at -20° and -80°C (≤ 20% and ≤ 15%, respectively). The SDS-AGE profiles revealed no visual changes regardless of duration of storage for specimens stored at room temperature, 4°C, and -80°C, except for 1 profile after 360 days at -80°C. Repeated freeze-thaw cycles did not affect SDS-AGE profiles. Appearance or strengthening of high-molecular-weight bands that could alter interpretation was evident in SDS-AGE profiles after storage at -20°C for ≥ 15 days (31/40 dogs). CONCLUSIONS AND CLINICAL RELEVANCE Storage of urine at -20° or -80°C for up to 1 year influenced the UPC without affecting clinical interpretation. Storage of urine specimens at -20°C impaired visual analysis of SDS-AGE. When SDS-AGE cannot be performed on fresh or recently refrigerated urine specimens, storage at -80°C is recommended.
目的 研究储存条件对犬尿蛋白肌酐比值(UPC)以及尿蛋白十二烷基硫酸钠 - 琼脂糖凝胶电泳(AGE)的影响。
样本 来自20只蛋白尿(UPC > 0.5)犬和20只非蛋白尿(UPC ≤ 0.2)犬的尿液样本。
步骤 对分别储存在室温(20°C)和4°C长达5天以及-20°C和-80°C长达360天的尿液样本进行UPC和SDS - AGE检测;部分样本进行3次冻融循环。将结果与新鲜尿液样本的结果进行比较。
结果 UPC不受室温储存或冷冻的影响。4°C储存5天后,非蛋白尿犬的样本UPC出现下降(10%);-20°C和-80°C储存90天后,两组样本的UPC均下降(分别≤ 20%和≤ 15%)。对于储存在室温、4°C和-80°C的样本,无论储存时间长短,SDS - AGE图谱均未显示明显变化,但-80°C储存360天后有1个图谱除外。反复冻融循环不影响SDS - AGE图谱。-20°C储存≥ 15天后,SDS - AGE图谱中出现或增强的高分子量条带可能会改变解读结果(40只犬中有31只)。
结论及临床意义 -20°C或-80°C储存尿液长达1年虽影响UPC,但不影响临床解读。-20°C储存尿液样本会损害SDS - AGE的视觉分析。当无法对新鲜或近期冷藏的尿液样本进行SDS - AGE检测时,建议在-80°C储存。