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摘要: 医院住院患者常常发生血液病原菌感染,死亡率高,早期快速鉴定病原体给予适当的抗微生物疗法是降低死亡率的关键。 血培养是诊断血液病原菌感染的金标准,然而血培养不仅耗时且阳性率低。 分子生物学方法可大幅度缩减诊断时间,提高诊断的灵敏度与特异性,为临床合理用药与准确治疗提供可靠的依据,进而提高患者的生存率。 本研究就血源性病原菌诊断的分子生物学方法(主要包括核酸杂交技术、核酸扩增技术、荧光实时定量PCR技术、DNA微阵列与基质辅助激光解析电离飞行时间质谱等技术)等进行综述。Abstract: Blood borne pathogen infection often occurs in hospitalized patients with high mortality rate, the early and rapid identification of pathogens and appropriate antimicrobial treatment are the key measures to reduce the mortality. Blood culture is the gold standard in the diagnosis of blood borne pathogen infection, blood culture, however, is not only time-consuming but also non-sensitive with low positive rate. Molecular biology method can greatly reduce diagnosis time, improve the sensitivity and specificity of diagnosis and provide reliable evidence for the rational clinical drug use and accurate treatment, which can greatly improve the patient's survival rate. This paper summarizes the progress in the research of molecular biology methods for the diagnosis of blood-borne pathogens, including nucleic acid hybridization technique, nucleic acid amplification technique, fluorescence real-time quantitative PCR, DNA microarray and matrix assisted laser desorption/ionizaton time of flight mass spectrometry(MALDI-TOF-MS).
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表 1 基于血培养阳性样本病原菌检测的商业方法
Table 1. Commercially available methods for pathogen detection in positive blood culture samples
仪器及生产厂家 技术原理 病原菌 灵敏度(%) 特异度(%) 诊断时间(h) Prove-itTM Sepsis(Mobidiag® Finland) 多重PCR、基因芯片 60种细菌、13种真菌 96.0 ~ 99.0 98.0 ~ 100.0 18 FilmArray®(BioFire Diagnostics,USA) 多重实时PCR 8种革兰阳性菌、11种革兰阴性菌、5种真菌 80.0 ~ 94.0 95.0 ~ 100.0 16 Verigene®(Nanosphere Technology,USA)Gram-negative bacteria(BC-GN) 基因芯片杂交 9种革兰阴性菌 89.0 ~ 100.0 93.0 ~ 100.0 17.5 Verigene®(Nanosphere Technology,USA)Gram-positive bacteria(BC-GP) 基因芯片杂交 13种革兰阳性菌 93.0 ~ 100.0 95.0 ~ 100.0 17.5 表 2 全血中直接检测病原菌的商业方法
Table 2. Commercially available methods for direct pathogen detection in whole blood samples
仪器及生产厂家 技术原理 病原菌 检测限
(CFU/ml)灵敏度
(%)特异度
(%)诊断时间
(h)T2Candida® test(T2Biosystems inc) PCR、核磁共振技术 5种念珠菌 1 100.0 98.0 3 SeptiFast(Roche Diagnostics,Germany) 实时PCR 25种细菌和真菌 3 ~ 30 68.0 ~ 75.0 86.0 ~ 92.0 6 SepsiTest(Molzym,Germany) PCR、测序 345种细菌和真菌 20 ~ 460 86.0 ~ 87.0 83.0 ~ 85.0 8 ~ 12 Vyoo®(SIRS,Germany) PCR、电泳 34种细菌、6种真菌 5 ~ 100 60.0 70.0 ~ 75.0 7 MagicplexTM(Seegene,Korea) 实时PCR 85种细菌、6种真菌 − 65.0 92.0 6 -
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2019-0073 血源性病原菌早期诊断技术研究进展.docx
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