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通过广谱实时聚合酶链反应诊断脑室引流相关细菌性脑膜炎

Diagnosis of ventricular drainage-related bacterial meningitis by broad-range real-time polymerase chain reaction.

作者信息

Deutch Susanna, Dahlberg Daniel, Hedegaard Jesper, Schmidt Michael B, Møller Jens K, Ostergaard Lars

机构信息

Department of Infectious Diseases, Skejby Sygehus, Aarhus University Hospital, Aarhus, Denmark.

出版信息

Neurosurgery. 2007 Aug;61(2):306-11; discussion 311-2. doi: 10.1227/01.NEU.0000255526.34956.E4.

Abstract

OBJECTIVE

To compare a broad-range real-time polymerase chain reaction (PCR) diagnostic strategy with culture to evaluate additional effects on the etiological diagnosis and the quantification of the bacterial load during the course of ventricular drainage-related bacterial meningitis (VR-BM).

METHODS

We applied a PCR that targeted conserved regions of the 16S ribosomal ribonucleic acid gene to cerebrospinal fluid (CSF) samples from patients with external ventricular drainage or a ventriculoperitoneal shunt during the course of VR-BM. We compared the PCR results with CSF cultures. A total of 350 routine CSF samples were consecutively collected from 86 patients. The CSF deoxyribonucleic acid was automatically purified and subjected to PCR. Amplicons from the PCR samples that were positive for VR-BM were subsequently deoxyribonucleic acid sequenced for final identification. Clinical data were extracted from patient files.

RESULTS

Sixteen patients had at least one VR-BM-positive sample as diagnosed from culture or PCR. Nineteen episodes were diagnosed with signs of VR-BM (n = 16 patients) or were determined to be contaminated (n = 3 patients). Four episodes of VR-BM were diagnosed via PCR alone and were predominantly caused by gram-negative pathogens, five episodes were diagnosed via culture alone, and seven episodes were diagnosed via both culture and PCR. Five patients had mixed infections. Overall, 71 samples were positive for VR-BM as indicated by either one or both of the methods. Eighteen CSF samples were VR-BM positive as indicated by culture alone, and 21 CSF samples were positive as indicated via PCR alone.

CONCLUSIONS

Culture supplemented with broad-range, real-time PCR may increase the number of etiologically diagnosed VR-BM episodes, particularly when these are caused by gram-negative bacteria.

摘要

目的

比较一种广谱实时聚合酶链反应(PCR)诊断策略与培养法,以评估其对脑室引流相关细菌性脑膜炎(VR-BM)病程中病因诊断及细菌载量定量的额外影响。

方法

我们将针对16S核糖体核糖核酸基因保守区域的PCR应用于VR-BM病程中外侧脑室引流或脑室腹腔分流患者的脑脊液(CSF)样本。我们将PCR结果与CSF培养结果进行比较。连续从86例患者中收集了350份常规CSF样本。CSF脱氧核糖核酸自动纯化后进行PCR。随后对VR-BM呈阳性的PCR样本的扩增子进行脱氧核糖核酸测序以进行最终鉴定。从患者病历中提取临床数据。

结果

16例患者至少有一份样本经培养或PCR诊断为VR-BM阳性。19次发作被诊断为有VR-BM体征(n = 16例患者)或被确定为污染(n = 3例患者)。4次VR-BM发作仅通过PCR诊断,主要由革兰氏阴性病原体引起,5次发作仅通过培养诊断,7次发作通过培养和PCR均诊断出来。5例患者有混合感染。总体而言,两种方法中的一种或两种均显示71份样本VR-BM呈阳性。仅培养显示18份CSF样本VR-BM呈阳性,仅PCR显示21份CSF样本呈阳性。

结论

广谱实时PCR辅助培养可能会增加病因诊断的VR-BM发作次数,特别是当这些发作由革兰氏阴性菌引起时。

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