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| 质谱直接鉴定血流感染病原菌前处理方法探索 |
| Exploration of pre-treatment methods for direct identification of bloodstream infection pathogens by Mass Spectrometry |
| 投稿时间：2025-12-15 修订日期：2026-04-02 |
| DOI： |
| 中文关键词: 基质辅助激光解吸电离飞行时间质谱 血流感染 直接鉴定 前处理方法 |
| 英文关键词: Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry Bloodstream infections Direct identification Pretreatment method |
| 基金项目:济宁医学院贺林院士新医学临床转化工作站科研基金 |
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| 中文摘要: |
| 目的 对比分离胶分离法与裂解法处理后的阳性血培养标本质谱鉴定的准确率，寻找最优前处理方法，助力临床血流感染快速精准诊疗。方法 收集实验室348份血培养阳性标本，分别采用分离胶分离法和裂解法对阳性培养液进行前处理后，直接用质谱检测。以培养菌落自动化鉴定药敏仪鉴定结果为标准评估直接质谱鉴定结果的准确性，无法鉴定或鉴定结果与镜下形态不符者进行16S r DNA基因测序为最终对照标准。并用模拟念珠菌样本验证优势方法的可靠性。结果 分离胶分离法中符合率高的甲酸法（89.37%）鉴定准确率低于裂解法中符合率低的红细胞裂解液法（95.69%），差异有统计学意义(P﹤0.05)。用直接鉴定较难的模拟念珠菌样本验证裂解法的准确率，配套试剂法符合率（88.39%）高于红细胞裂解液法（86.61%），但差异无统计学意义（P﹥0.05）。结论 红细胞裂解液法是最优的前处理方法，准确率高，经济实用性强，可以为血流感染的早期精准诊断提供参考依据。 |
| 英文摘要: |
| Objective Comparing the accuracy of mass spectrometry identification for positive blood culture samples processed by separator gel method versus lysis method, to identify the optimal pretreatment approach, thereby facilitating rapid and precise clinical diagnosis and treatment of bloodstream infections. Methods A total of 348 positive blood culture samples were collected in the laboratory. After pretreatment of the positive culture broth using the separator gel method and the lysis method, respectively, direct mass spectrometry detection was performed. The accuracy of the direct mass spectrometry identification results was evaluated against the automated colony identification and drug susceptibility instrument results as the standard. For samples that could not be identified or whose identification results were inconsistent with microscopic morphology, 16S rDNA gene sequencing was performed as the final reference standard. Additionally, simulated Candida samples were used to verify the reliability of the superior method. Results In the separator gel method, the formic acid approach, which demonstrated a high concordance rate (89.37%), showed a lower identification accuracy compared to the red blood cell lysis method (95.69%) in the lysis method, despite the latter having a lower concordance rate. The difference was statistically significant (P < 0.05). To further validate the accuracy of the lysis method, simulated Candida samples—known for being challenging in direct identification—were tested. The commercial kit method achieved a higher concordance rate (88.39%) than the red blood cell lysis method (86.61%), but the difference was not statistically significant (P > 0.05).Conclusion The red blood cell lysis method is the optimal pretreatment approach, offering high accuracy and strong cost-effectiveness, and can serve as a reference for the early and precise diagnosis of bloodstream infections. |
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