干血斑技术用于重症感染患者美罗培南精准给药的评价研究

    Evaluation Study of Dried Blood Spot Technology for Guiding Precision Dosing of Meropenem in Critically Ill Patients with Severe Infection

    • 摘要:
      目的  建立一种基于干血斑(dried blood spot,DBS)技术的美罗培南血药浓度检测方法,并探讨其在重症感染患者精准用药中的应用价值。
      方法 采用HPLC测定DBS中美罗培南的浓度,优化样品前处理流程和色谱条件,进行完整的方法学验证(包括特异性、线性、准确度、精密度、提取回收率、基质效应和稳定性),并与传统静脉血浆样本进行一致性比较(Bland-Altman法和Passing-Bablok回归法),验证重症感染患者样本检验临床实用性。
      结果 所建立的DBS-HPLC在1~100 μg·mL−1线性良好(R2=0.9982),定量下限为1 μg·mL−1。特异性、准确度、精密度、提取回收率、基质效应和稳定性均符合生物样本分析要求,DBS技术在常温、4 ℃ 和−20 ℃条件下均能使美罗培南保持72 h的稳定性,且在常温下的稳定性优于血浆样本。DBS采样与传统静脉全血采样测得的美罗培南血药浓度结果具有良好的一致性。临床浓度监测显示,以DBS法检测结合传统静脉血浆法复核,98%(49/50)的患者美罗培南谷浓度达到目标阈值。
      结论 DBS-HPLC可用于美罗培南血药浓度的快速、微创监测,为重症感染患者的个体化给药提供可靠支持。

       

      Abstract:
      OBJECTIVE To develop an assay for determining meropenem blood concentrations using dried blood spot(DBS) technology and to evaluate its application value in guiding precision dosing for critically ill patients with severe infection.
      METHODS Meropenem concentrations in DBS samples were quantified using HPLC. The sample pretreatment procedures and chromatographic conditions were optimized, followed by a full method validation including specificity, linearity, accuracy, precision, extraction recovery, matrix effects, and stability. The agreement between meropenem concentrations measured in DBS and paired conventional venous plasma samples was assessed using Bland-Altman and Passing-Bablok regression analyses. Clinical utility of testing samples from critically ill patients with severe infection was verified.
      RESULTS The established DBS-HPLC demonstrated excellent linearity over the concentration range of 1−100 μg·mL−1(R2=0.9982), with a lower limit of quantification of 1 μg·mL−1. The specificity, accuracy, precision, extraction recovery, matrix effects, and stability all met the acceptance criteria for bioanalytical method validation. DBS technology maintained the stability of meropenem for 72 h under ambient temperature, 4 ℃, and −20 ℃ conditions, with superior stability at ambient temperature compared to plasma samples. Meropenem blood concentration results obtained from DBS sampling demonstrated good consistency with those from traditional venous plasma sampling. Clinical monitoring showed that 98% (49/50) of patients achieved the target trough concentration of meropenem based on DBS analysis verified by conventional venous plasma testing.
      CONCLUSION DBS-HPLC enables rapid and minimally-invasive monitoring of meropenem blood concentration, offering reliable support for personalized antimicrobial therapy in critically ill patients with severe infection.

       

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