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首页> 外文期刊>Journal of Clinical Microbiology >Pathogen Identification by Multiplex LightMix Real-Time PCR Assay in Patients with Meningitis and Culture-Negative Cerebrospinal Fluid Specimens
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Pathogen Identification by Multiplex LightMix Real-Time PCR Assay in Patients with Meningitis and Culture-Negative Cerebrospinal Fluid Specimens

机译：脑膜炎和培养阴性的脑脊液标本的多重LightMix实时PCR测定病原体。

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ABSTRACT Acute bacterial meningitis is a medical emergency, and delays in initiating effective antimicrobial therapy result in increased morbidity and mortality. Culture-based methods, thus far considered the “gold standard” for identifying bacterial microorganisms, require 24 to 48 h to provide a diagnosis. In addition, antimicrobial therapy is often started prior to clinical sample collection, thereby decreasing the probability of confirming the bacterial pathogen by culture-based methods. To enable a fast and accurate detection of the most important bacterial pathogens causing meningitis, namely, Streptococcus pneumoniae , Haemophilus influenzae , Neisseria meningitidis , Streptococcus agalactiae , and Listeria monocytogenes , we evaluated a commercially available multiplex LightMix real-time PCR (RT-PCR) in 220 cerebrospinal fluid (CSF) specimens. The majority of CSF samples were collected by lumbar puncture, but we also included some CSF samples from patients with symptoms of meningitis from the neurology department that were recovered from shunts. CSF samples were analyzed by multiplex RT-PCR enabling a first diagnosis within a few hours after sample arrival at our institute. In contrast, bacterial identification took between 24 and 48 h by culture. Overall, a high agreement of bacterial identification between culture and multiplex RT-PCR was observed (99%). Moreover, multiplex RT-PCR enabled the detection of pathogens, S. pneumoniae ( n = 2), S. agalactiae ( n = 1), and N. meningitidis ( n = 1), in four culture-negative samples. As a complement to classical bacteriological CSF culture, the LightMix RT-PCR assay proved to be valuable by improving the rapidity and accuracy of the diagnosis of bacterial meningitis.
机译：摘要急性细菌性脑膜炎是一种医疗急症，延迟开始有效的抗菌治疗会增加发病率和死亡率。迄今为止，基于培养的方法被认为是鉴定细菌微生物的“金标准”，需要24至48小时才能做出诊断。另外，抗微生物治疗通常在临床样品收集之前开始，从而降低了通过基于培养物的方法确认细菌病原体的可能性。为了能够快速，准确地检测出引起脑膜炎的最重要细菌病原体，即肺炎链球菌，流感嗜血杆菌，脑膜炎奈瑟菌，无乳链球菌和单核细胞增生李斯特菌，我们评估了市售的多重LightMix实时PCR（RT-PCR）在220例脑脊液（CSF）标本中。大部分CSF样本是通过腰椎穿刺收集的，但我们还包括了从分流术中回收的神经科脑膜炎症状患者的一些CSF样本。通过多重RT-PCR对脑脊液样品进行了分析，能够在样品到达我们研究所后的几个小时内进行首次诊断。相反，通过培养，细菌鉴定花费了24至48小时。总体而言，在培养物和多重RT-PCR之间观察到细菌鉴定的一致性很高（99％）。此外，多重RT-PCR能够在四个培养阴性样品中检测到病原体，如肺炎链球菌（n = 2），无乳链球菌（n = 1）和脑膜炎奈瑟菌（n = 1）。作为经典细菌CSF培养的补充，LightMix RT-PCR测定通过提高诊断细菌性脑膜炎的速度和准确性被证明是有价值的。

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