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首页> 外文期刊>Journal of Clinical Microbiology >Diagnosis of Listeria monocytogenes Meningoencephalitis by Real-Time PCR for the hly Gene
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Diagnosis of Listeria monocytogenes Meningoencephalitis by Real-Time PCR for the hly Gene

机译：hly基因实时荧光定量PCR诊断单核细胞增生李斯特菌脑膜脑炎

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Listeria monocytogenes is a bacterial pathogen that can invade the central nervous system (CNS), causing meningoencephalitis and brain abscesses. The diagnosis of CNS listeriosis, based on the isolation of the bacteria in the cerebrospinal fluid (CSF), can be difficult because of previous antibiotic treatment and a low number of bacteria in the CSF. To improve the sensitivity of microbiological diagnosis, we have developed a real-time PCR assay for detecting and quantifying L. monocytogenes DNA in the CSF. The designed primers specifically amplify the L. monocytogenes hly gene, which encodes listeriolysin O, a pore-forming cytolysin. The PCR assay for the hly gene (PCR-hly) provides reproducible quantitative results over a wide dynamic range of concentrations and was highly sensitive while detecting a single gene copy/ml. By assaying a large panel of bacterial species, including species secreting pore-forming cytolysin, we determined the specificity of the PCR-hly, which exclusively detects the L. monocytogenes DNA. We then analyzed 214 CSF samples from patients suspected of having CNS listeriosis. PCR-hly was positive in all cases in which L. monocytogenes was isolated by culture. Positive PCR-hly of the CSF was also obtained for five additional, clinically confirmed cases of CNS listeriosis for which bacterial cultures were negative presumably due to previous treatment with antibiotics. As a complement to classical bacteriological CSF culture, our designed real-time PCR-hly assay proved to be valuable by enhancing the rapidity and the accuracy of the diagnosis of CNS infection by L. monocytogenes. In addition, the quantitative results provided may, in some instances, be useful for the follow-up of patients under treatment.
机译：单核细胞增生李斯特菌是一种细菌性病原体，可侵袭中枢神经系统（CNS），引起脑膜脑炎和脑脓肿。由于以前的抗生素治疗以及脑脊液中细菌数量少，因此基于脑脊液（CSF）中细菌分离的中枢神经系统李斯特菌病的诊断可能很困难。为了提高微生物诊断的敏感性，我们开发了一种实时PCR检测方法，用于检测和定量脑脊液中的单核细胞增生李斯特菌DNA。设计的引物可特异性扩增单核细胞增生李斯特菌 hly 基因，该基因编码利斯特菌溶血素O（一种形成孔的细胞溶素）。 hly 基因的PCR测定法（PCR-hly）可在宽动态范围内提供可重复的定量结果，并且在检测单个基因拷贝/ ml时非常灵敏。通过分析一大批细菌物种，包括分泌孔形成性溶菌素的物种，我们确定了PCR-hly的特异性，它专门检测单核细胞增生李斯特菌DNA。然后，我们从疑似中枢神经系统李斯特菌病患者中分析了214份CSF样本。在通过培养分离出单核细胞增生李斯特菌的所有情况下，PCR-hly均为阳性。对于另外五例经临床证实的中枢神经系统李斯特菌病病例，也获得了CSF的阳性PCR阳性结果，推测这些病例的细菌培养可能是由于先前使用抗生素治疗而阴性。作为经典细菌CSF培养的补充，我们设计的实时PCR-hly检测通过提高单核细胞增生李斯特氏菌诊断中枢神经系统感染的速度和准确性被证明是有价值的。另外，在某些情况下，提供的定量结果可能对治疗中患者的随访有用。

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