Puigvehí Marc, Hernández Juanjo, Broquetas Teresa, Coll Susanna, Garcia-Retortillo Montserrat, Cañete Nuria, Giménez Maria Dolors, Garcia Mar, Bory Felipe, Salvadó Margarita, Solà Ricard, Carrión José A
Liver Section, Gastroenterology Department, Hospital del Mar, Universitat Autònoma de Barcelona.
IMIM (Hospital del Mar Medical Research Institute), Barcelona, Spain.
PLoS One. 2016 Dec 16;11(12):e0164883. doi: 10.1371/journal.pone.0164883. eCollection 2016.
INTRODUCTION & AIMS: Cryopreservation of serum samples is a standard procedure for biomedical research in tertiary centers. However, studies evaluating the long-term biological stability of direct liver fibrosis markers using cryopreserved samples are scarce.
We compared the stability of hyaluronic acid (HA), tissue inhibitor of metalloproteinases (TIMP-1) and amino-terminal propeptide of type III procollagen (PIIINP) in 225 frozen serum samples of HCV-infected patients with a paired liver biopsy for up to 25 years (1990-2014). Moreover, we assessed the diagnostic accuracy (AUROC) of the Enhanced Liver Fibrosis (ELF®) score to identify significant fibrosis (F2-4) and its predictive capacity to identify clinical events during follow-up.
Seventy-six patients (39,8%) had mild fibrosis (F0-1) and 115 (60,2%) significant fibrosis (F2-4). HA, PIIINP and TIMP-1 values remained stable during the period from 1995 to 2014 while those of 1990-94 were slightly higher. We did not find significant differences in the median ELF® values during the 20-year period from 1995-2014 in patients with mild (from 8,4 to 8,7) and significant fibrosis (from 9,9 to 10,9) (p = ns between periods and fibrosis stages). The AUROCs of ELF® to identify significant fibrosis were high in all the periods (from 0,85 to 0,91). The ELF® score showed a good predictive capability to identify clinical events during follow-up.
The biological stability of direct serum markers (HA, PIIINP and TIMP-1) using HCV-infected samples cryopreserved for 20 years is good. Therefore, the diagnostic accuracy of the ELF® score to identify significant fibrosis and clinical events during follow-up is very high.
血清样本的冷冻保存是三级医疗中心生物医学研究的标准程序。然而，使用冷冻保存样本评估直接肝纤维化标志物长期生物学稳定性的研究较少。
我们比较了225份丙型肝炎病毒（HCV）感染患者的冷冻血清样本中透明质酸（HA）、金属蛋白酶组织抑制剂（TIMP - 1）和III型前胶原氨基末端前肽（PIIINP）的稳定性，并对其进行了长达25年（1990 - 2014年）的配对肝活检。此外，我们评估了增强肝纤维化（ELF®）评分诊断显著纤维化（F2 - 4）的准确性（受试者工作特征曲线下面积，AUROC）及其在随访期间识别临床事件的预测能力。
76例患者（39.8%）有轻度纤维化（F0 - 1），115例（60.2%）有显著纤维化（F2 - 4）。HA、PIIINP和TIMP - 1值在1995年至2014年期间保持稳定，而1990 - 1994年的值略高。在1995 - 2014年的20年期间，轻度纤维化患者（从8.4到8.7）和显著纤维化患者（从9.9到10.9）的ELF®中位数无显著差异（不同时期和纤维化阶段之间p =无显著性差异）。ELF®在所有时期诊断显著纤维化的AUROC都很高（从0.85到0.91）。ELF®评分在随访期间显示出良好的识别临床事件的预测能力。
使用冷冻保存20年的HCV感染样本，直接血清标志物（HA、PIIINP和TIMP - 1）的生物学稳定性良好。因此，ELF®评分在诊断显著纤维化和随访期间临床事件方面的准确性非常高。
