Marín-Romero Antonio, Di Zeo-Sánchez Daniel E, Tabraue-Chávez Mavys, Villanueva-Paz Marina, Pinazo-Bandera Jose M, Sanabria-Cabrera Judith, García-Cortés Miren, Díaz-Mochón Juan J, Lucena M Isabel, Andrade Raúl J, Stephens Camilla, Pernagallo Salvatore
DESTINA Genomica S.L., Edificio BIC, Parque Tecnológico Ciencias de la Salud, Granada, Spain.
Servicios de Aparato Digestivo y Farmacología Clínica, Instituto de Investigación Biomédica de Málaga y Plataforma en Nanomedicina-IBIMA Plataforma, Hospital Universitario Virgen de la Victoria, Universidad de Málaga, Málaga, Spain.
PLoS One. 2025 May 30;20(5):e0324858. doi: 10.1371/journal.pone.0324858. eCollection 2025.
There is currently a need to investigate new biomarkers of acute liver injury (ALI) that are highly specific to the liver and capable of detecting early-stage liver damage. In this regard, circulating microRNAs (miRNAs), particularly microRNA122 (miRNA122), have recently been proposed as promising new candidate biomarkers. However, the quantification of miRNAs is not a standardized technique and presents several challenges that hinder its routine use. The aim of this work is to validate the innovative Dynamic Chemical Labeling (DCL) PCR-Free technology for its ability to detect miRNA122 in patient samples using Luminex xMAP platforms. The DCL PCR-Free technology was used to directly measure and monitor miRNA122 levels in serum samples from patients with ALI. Patients were monitored throughout the recovery process from liver injury, from the time of detection and for up to 30 days afterwards, with follow-up over three separate visits. The results demonstrate the robustness of the method, with sensitivity of 3.36% and precision of 99.80%, and show a high correlation between miRNA122 and traditional liver injury markers such as ALT (r = 0.8150, p = 0.0001), AST (r = 0.7895, p = 0.0002) and TBL (r = 0.2646, p = 0.3406) throughout the ALI recovery process. In conclusion, measuring miRNA122 levels using the DCL PCR-Free method provides a distinctive approach, not only for diagnosing ALI patients but also for effectively monitoring disease progression, tracking liver recovery, and evaluating treatment effectiveness.
目前需要研究急性肝损伤(ALI)的新型生物标志物,这些标志物对肝脏具有高度特异性,并且能够检测早期肝损伤。在这方面,循环微RNA(miRNA),特别是微RNA122(miRNA122),最近被提议作为有前景的新型候选生物标志物。然而,miRNA的定量并非标准化技术,存在若干阻碍其常规应用的挑战。本研究的目的是验证创新的无PCR动态化学标记(DCL)技术利用Luminex xMAP平台检测患者样本中miRNA122的能力。采用无PCR DCL技术直接测量和监测急性肝损伤患者血清样本中的miRNA122水平。在患者从肝损伤开始恢复的整个过程中进行监测,从检测之时起直至之后30天,分三次单独随访进行跟踪。结果证明了该方法的稳健性,灵敏度为3.36%,精密度为99.80%,并且在急性肝损伤恢复过程中,miRNA122与传统肝损伤标志物如谷丙转氨酶(ALT,r = 0.8150,p = 0.0001)、谷草转氨酶(AST,r = 0.7895,p = 0.0002)和总胆红素(TBL,r = 0.2646,p = 0.3406)之间呈现高度相关性。总之,使用无PCR DCL方法测量miRNA122水平提供了一种独特的方法,不仅可用于诊断急性肝损伤患者,还可有效监测疾病进展、跟踪肝脏恢复情况以及评估治疗效果。