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基于基质辅助激光解吸电离飞行时间质谱系统的血流感染致病菌早期检测
作者:张可昕 胡翀 陈琛 何赏 杨明 王成彬  
单位:325000 浙江温州 温州医科大学检验医学院/生命科学学院(张可昕、杨明、王成彬) 100853 北京 解放军总医院临床检验科(张可昕、胡翀、陈琛、何赏、杨明、王成彬) 
关键词:基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS) 细菌性血流感染 早期诊断 
分类号:R03;R373.13
出版年,卷(期):页码:2018,43(1):17-22
摘要:

[摘要] 目的 基于基质辅助激光解吸电离飞行时间质谱系统(MALDI-TOF MS)建立早期血流感染致病菌检测体系。方法 选取大肠埃希菌、铜绿假单胞菌、鲍曼不动杆菌、肺炎克雷伯杆菌、屎肠球菌、粪肠球菌、金黄色葡萄球菌、表皮葡萄球菌、变形杆菌、肺炎链球菌等10种临床最常见的病原菌作为检测对象,结合多重PCR技术和Mass ARRAY iPLEX单碱基延伸技术、MALDI-TOF MS技术,模拟细菌感染血液样本建立MALDI-TOF MS细菌核酸检测体系,测定其灵敏度;选取201735月在解放军总医院急诊科收集的疑似脓毒症患者血液样本33例对该体系进行验证,并将检测结果与微生物科血培养结果进行比较。结果 本研究所建立的MALDI-TOF质谱血液致病菌检测系统可同时检测分属两组的10种细菌,所有细菌样本均可见延伸峰;灵敏度为100CFU/ml;与33例临床血培养鉴定结果的阴性符合率为100%(27/27)6例血培养阳性样品中有2例为摩氏摩根菌和人葡萄球菌感染,因这两种细菌不包含在10种检测对象之内而未能通过质谱检出,其余4例血培养阳性样品与质谱检测结果的符合率为100%结论 本研究建立的 MALDI-TOF质谱检测体系可从全血样本中直接检测10种临床常见血流感染致病菌,无需进行培养,与传统血培养及生化鉴定相比具有耗时短、灵敏度高、准确度高等优势,可在一定程度上满足临床早期、快速诊断血流感染致病菌的需求。

[Abstract] Objective To establish a clinical method for early detection of the bloodstream infection bacteria based on matrix-assisted laser ionization time of flight mass spectrometry (MALDI-TOF MS). Methods After consulting the Laboratory Information System statistics and domestic related literature, We chose 10 kinds of bacteria (Escherichia coli., Pseudomonas aeruginosa, Acinetobacter baumannii, Klebsiella pneumoniae, Enterococcus faecium, Enterococcus faecalis, Staphylococcus aureus and Staphylococcus epidermidis, Proteus mirabilis, Streptococcus pneumoniae) as target. The MALDI-TOF MS was established using simulated bacterial infection blood samples. From March to May 2017, 33 blood samples of suspected sepsis patients from Emergency Department, General Hospital of PLA, were tested. Results The MALDI-TOF MS whose detecting sensitivity was 100CFU/ml had the same negative detection rate with the blood culture (27cases/27cases, 100%). Besides the 2 samples of Morganella morganii infection and Staphylococcus hominis infection were out of the range, the results of the remaining 4 positive samples were consistent (100%). Conclusion Compared to the blood culture and biochemical identification, MALDI-TOF MS can rapidly detect 10 kinds of bloodstream infection bacteria with high sensitivity and high accuracy.

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