多重聚合酶链反应在细菌性脑膜炎诊断中的应用。
Multiplex-PCR for diagnosis of bacterial meningitis.
机构信息
Faculdade de Ciências Farmacêuticas, Department of Clinical Chemistry and Toxicology, Universidade de São Paulo (USP), São Paulo, Brazil.
Biomedical Sciences Institute, Vaccine Development Laboratory, Department of Microbiology, USP, São Paulo, Brazil.
出版信息
Braz J Microbiol. 2019 Apr;50(2):435-443. doi: 10.1007/s42770-019-00055-9. Epub 2019 Feb 22.
Considering the great lethality and sequels caused by meningitis, rapid diagnosis and prompt treatment initiation have a great impact on patient outcome. Here, we developed a multiplex-PCR for simultaneous detection of the four most prevalent bacterial pathogens directly in CSF samples. The multiplex-PCR was designed to detect the following genes: fbsA (Streptococcus agalactiae), lytA (Streptococcus pneumoniae), crtA (Neisseria meningitidis), p6 (Haemophilus influenzae), and 16S rRNA (any bacterial agent). The multiplex-PCR showed a DNA detection limit of 1 pg/μL. Among 447 CSF samples tested, 40 were multiplex-PCR positive, in which 27 and 13 had positive and negative bacterial culture, respectively. Our multiplex-PCR is fast, reliable, and easily implementable into a laboratory routine for bacterial meningitis confirmation, especially for patients who previously started antimicrobial therapy. Our molecular approach can substantially improve clinical diagnosis and epidemiological measures of meningitis disease burden.
鉴于脑膜炎的巨大杀伤力和后遗症，快速诊断和及时开始治疗对患者的预后有很大影响。在这里，我们开发了一种多重 PCR 方法，可直接从 CSF 样本中同时检测四种最常见的细菌病原体。该多重 PCR 设计用于检测以下基因：fbsA（无乳链球菌）、lytA（肺炎链球菌）、crtA（脑膜炎奈瑟菌）、p6（流感嗜血杆菌）和 16S rRNA（任何细菌）。多重 PCR 的 DNA 检测限为 1 pg/μL。在检测的 447 份 CSF 样本中，40 份为多重 PCR 阳性，其中 27 份和 13 份分别有阳性和阴性细菌培养。我们的多重 PCR 快速、可靠，易于在实验室常规中实施，用于细菌脑膜炎的确认，特别是对于那些之前开始抗菌治疗的患者。我们的分子方法可以大大提高脑膜炎疾病负担的临床诊断和流行病学措施。
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