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首页> 外文期刊>Journal of Clinical Microbiology >Rapid Diagnosis of Bacterial Meningitis by Real-Time PCR and Fluorescence In Situ Hybridization
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Rapid Diagnosis of Bacterial Meningitis by Real-Time PCR and Fluorescence In Situ Hybridization

机译：实时PCR和荧​​光原位杂交技术快速诊断细菌性脑膜炎

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Real-time PCR and fluorescence in situ hybridization (FISH) were evaluated as rapid methods for the diagnosis of bacterial meningitis and compared to standard diagnostic procedures. For PCR, a LightCycler approach was chosen, implementing eubacterial and specific PCR assays for the most relevant bacteria. For FISH, a similar probe set containing eubacterial and specific probes was composed of published and newly designed probes. Both methods were evaluated by use of cerebrospinal fluid (CSF) samples from patients with suspected bacterial meningitis. For all microscopy- and culture-positive samples (n = 28), the eubacterial PCR was positive. In addition, all identifiable pathogens were detected with specific PCR assays, according to an algorithm based on the Gram stain. The FISH method detected the pathogen in 13 of 18 positive samples. While the FISH method remained negative for all microscopy- and culture-negative samples (n = 113), the eubacterial PCR was positive for five of these samples. Sequencing of the amplicon revealed the presence of Neisseria meningitidis, Streptococcus agalactiae, and Haemophilus influenzae in three of these five samples. In addition, samples with discordant results by culture and microscopy were successfully investigated by PCR (10 samples) and FISH (5 samples). In conclusion, PCR is a highly sensitive tool for rapid diagnosis of bacterial meningitis. FISH is less sensitive but is useful for the identification of CSF samples showing bacteria in the Gram stain. Based on our results, an approach for laboratory diagnosis of meningitis including PCR and FISH is discussed.
机译：实时PCR和荧​​光原位杂交（FISH）被评估为诊断细菌性脑膜炎的快速方法，并与标准诊断程序进行了比较。对于PCR，选择了LightCycler方法，对最相关的细菌实施真细菌和特异性PCR分析。对于FISH，包含真细菌和特定探针的相似探针组由已发布和新设计的探针组成。两种方法均通过使用疑似细菌性脑膜炎患者的脑脊液（CSF）样品进行评估。对于所有显微镜和培养阳性样本（ n = 28），真细菌PCR均为阳性。此外，根据基于革兰氏染色的算法，所有的可识别病原体均通过特定的PCR检测法检测到。 FISH方法在18个阳性样本中的13个中检测到病原体。尽管FISH方法在所有显微镜和培养阴性样品中均阴性（ n = 113），但其中5个样品的真细菌PCR阳性。扩增子的测序表明，在这五个样本中的三个样本中存在脑膜炎奈瑟氏菌，无乳链球菌和流感嗜血杆菌。此外，通过PCR（10个样品）和FISH（5个样品）成功地研究了培养和显微镜检查结果不一致的样品。总之，PCR是用于快速诊断细菌性脑膜炎的高度敏感的工具。 FISH的敏感性较低，但可用于鉴定显示革兰氏染色细菌的CSF样品。根据我们的结果，讨论了一种实验室诊断脑膜炎的方法，包括PCR和FISH。

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