Chapman Daniel P, Gooding Kim M, McDonald Timothy J, Shore Angela C
Diabetes and Vascular Research, University of Exeter, Exeter, UK.
National Institute for Health Research (NIHR) Exeter Clinical Research Facility, University of Exeter, Exeter, UK.
Pract Lab Med. 2019 Mar 22;15:e00120. doi: 10.1016/j.plabm.2019.e00120. eCollection 2019 May.
Increasing albumin to creatinine ratio (ACR) within the normal range is a risk factor for cardiovascular disease in the general population. Clinical and epidemiological studies often store urine samples for long durations prior to ACR assessment. The stability of ACR at the lowest urinary albumin concentrations during prolonged storage has not been previously studied because routine clinical assays can't quantify very low concentrations of albumin.
To determine the stability of urinary albumin and creatinine over 12 months in samples stored at -20 °C and -80 °C using an assay which enables assessment of previously undetectable levels of albumin and to investigate if additives can be used to prevent urinary albumin degradation.
ACR was measured in 30 urine samples from healthy subjects on the day of collection. Each sample was divided into 5 portions, each receiving a different treatment; alkalisation, protease inhibiter, boric acid, low protein binding tubes and no treatment (control). Samples were stored at -20 °C and -80 °C and ACR was analysed again after 12 months.
Mean (95% CI) percent change in ACR was -34.3% (-47.2 to -21.4; p < 0.0001) and -1.8% (-9.4 to 5.8; p = 0.91) in samples stored at -20 °C and -80 °C respectively. Treating samples did not prevent the reduction in albumin at -20 °C (p < 0.001).
The loss in urinary albumin concentration which occurs during storage at -20 °C for 12 months is not prevented by pre-treating samples prior to storage. For accurate determination of albumin concentration or ACR, samples should be stored at -80 °C on day of collection.
在普通人群中,正常范围内的白蛋白与肌酐比值(ACR)升高是心血管疾病的一个危险因素。临床和流行病学研究在评估ACR之前,通常会将尿液样本长时间保存。由于常规临床检测无法对极低浓度的白蛋白进行定量,因此此前尚未研究过在长时间保存过程中,最低尿白蛋白浓度下ACR的稳定性。
使用一种能够评估先前无法检测到的白蛋白水平的检测方法,确定在-20°C和-80°C下保存12个月的样本中尿白蛋白和肌酐的稳定性,并研究是否可以使用添加剂来防止尿白蛋白降解。
在收集当天,对30名健康受试者的尿液样本进行ACR测量。每个样本分成5份,每份接受不同的处理;碱化、蛋白酶抑制剂、硼酸、低蛋白结合管和不处理(对照)。样本分别保存在-20°C和-80°C,12个月后再次分析ACR。
分别保存在-20°C和-80°C的样本中,ACR的平均(95%CI)变化百分比分别为-34.3%(-47.2至-21.4;p<0.0001)和-1.8%(-9.4至5.8;p=0.91)。处理样本并不能防止在-20°C下白蛋白的减少(p<0.001)。
在-20°C下保存12个月期间发生的尿白蛋白浓度损失,不能通过在保存前对样本进行预处理来防止。为了准确测定白蛋白浓度或ACR,样本应在收集当天保存在-80°C。