Pfau Anja, Wytopil Monika, Chauhan Kinsuk, Reichel Martin, Coca Steve G, Aronson Peter S, Eckardt Kai-Uwe, Knauf Felix
Department of Nephrology and Medical Intensive Care, Charité - Universitätsmedizin Berlin, Berlin, Germany.
Department of Nephrology and Hypertension, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany.
Kidney Int Rep. 2020 Sep 2;5(11):2013-2020. doi: 10.1016/j.ekir.2020.08.029. eCollection 2020 Nov.
Alterations in oxalate homeostasis are associated with kidney stone disease and progression of chronic kidney disease (CKD). However, accurate measurement of plasma oxalate (P) concentrations in large patient cohorts is challenging as prompt acidification of samples has been deemed necessary. In the present study, we investigated the effects of variations in sample handling on P results and examined an alternative strategy to the established preanalytical procedures.
The effect of storage time at room temperature (RT) and maintenance of samples at -80°C was tested. P was measured in 1826 patients enrolled in the German Chronic Kidney Disease (GCKD) study, an ongoing multicenter, prospective, observational cohort study.
We demonstrate that P concentrations increased rapidly when samples were maintained at RT. This was most relevant for P <10 μM, as concentrations more than doubled within a few hours. Immediate freezing on dry ice and storage at -80°C provided stable results and allowed postponement of acidification for >1 year. In the patients of the lowest estimated glomerular filtration rate (eGFR) quartile, median P was 2.7 μM (interquartile range [IQR] <2.0-4.2) with a median eGFR of 25.1 ml/min per 1.73 m (IQR 20.3-28.1).
We conclude that immediate freezing and maintenance of plasma samples at -80°C facilitates the sample collection process and allows accurate P assessment in large cohorts. The present study may serve as a reference for sample handling to assess P in clinical trials and to determine its role in CKD progression.
草酸盐稳态的改变与肾结石疾病及慢性肾脏病(CKD)的进展相关。然而,在大型患者队列中准确测量血浆草酸盐(P)浓度具有挑战性,因为样本的快速酸化被认为是必要的。在本研究中,我们调查了样本处理方式的变化对P结果的影响,并研究了一种替代既定分析前程序的策略。
测试了室温(RT)下的储存时间以及在-80°C保存样本的影响。在德国慢性肾脏病(GCKD)研究中纳入的1826例患者中测量了P,该研究是一项正在进行的多中心、前瞻性、观察性队列研究。
我们证明,当样本在室温下保存时,P浓度迅速升高。这对于P<10μM的情况最为明显,因为浓度在数小时内增加了一倍多。立即在干冰上冷冻并在-80°C保存可提供稳定的结果,并允许将酸化推迟1年以上。在估计肾小球滤过率(eGFR)最低四分位数的患者中,P中位数为2.7μM(四分位间距[IQR]<2.0-4.2),eGFR中位数为每1.73 m² 25.1 ml/min(IQR 20.3-28.1)。
我们得出结论,血浆样本立即冷冻并在-80°C保存有助于样本采集过程,并允许在大型队列中准确评估P。本研究可为临床试验中评估P的样本处理以及确定其在CKD进展中的作用提供参考。