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针对神经外科术后细菌性脑膜炎的 16S 核糖体 RNA 的聚合酶链反应诊断。

Polymerase chain reaction targeting 16S ribosomal RNA for the diagnosis of bacterial meningitis after neurosurgery.

机构信息

Faculdade de Medicina (FMUSP), Universidade de Sao Paulo, Sao Paulo, SP, BR.

Laboratorio de Investigacao Medica 49, Bacteriologia, Sao Paulo SP, BR.

出版信息

Clinics (Sao Paulo). 2021 Jan 22;76:e2284. doi: 10.6061/clinics/2021/e2284. eCollection 2021.

Abstract

OBJECTIVES

Bacterial and aseptic meningitis after neurosurgery can present similar clinical signs and symptoms. The aims of this study were to develop and test a molecular method to diagnose bacterial meningitis (BM) after neurosurgery.

METHODS

A 16S ribosomal RNA gene PCR-based strategy was developed using artificially inoculated cerebrospinal fluid (CSF) followed by sequencing. The method was tested using CSF samples from 43 patients who had undergone neurosurgery and were suspected to suffer from meningitis, and from 8 patients without neurosurgery or meningitis. Patients were classified into five groups, confirmed BM, probable BM, possible BM, unlikely BM, and no meningitis.

RESULTS

Among the samples from the 51 patients, 21 samples (41%) were culture-negative and PCR-positive. Of these, 3 (14%) were probable BM, 4 (19%) were possible BM, 13 (62%) were unlikely BM, and 1 (5%) was meningitis negative. Enterobacterales, non-fermenters (Pseudomonas aeruginosa and Acinetobacter baumannii), Staphylococcus haemolyticus, Granulicatella, Variovorax, and Enterococcus cecorum could be identified. In the group of patients with meningitis, a good agreement (3 of 4) was observed with the results of cultures, including the identification of species.

CONCLUSION

Molecular methods may complement the diagnosis, guide treatment, and identify non-cultivable microorganisms. We suggest the association of methods for suspected cases of BM after neurosurgery, especially for instances in which the culture is negative.

摘要

目的

神经外科手术后的细菌性和无菌性脑膜炎可能表现出相似的临床体征和症状。本研究旨在开发和测试一种用于诊断神经外科手术后细菌性脑膜炎(BM)的分子方法。

方法

使用人工接种的脑脊液(CSF),建立了一种基于 16S 核糖体 RNA 基因 PCR 的策略,随后进行测序。该方法使用来自 43 名接受过神经外科手术且疑似患有脑膜炎的患者和 8 名无神经外科手术或脑膜炎的患者的 CSF 样本进行了测试。患者被分为五组,确诊 BM、可能 BM、可能 BM、不太可能 BM 和无脑膜炎。

结果

在 51 名患者的样本中,有 21 份(41%)培养结果为阴性但 PCR 结果为阳性。其中,3 份(14%)为可能 BM,4 份(19%)为可能 BM,13 份(62%)为不太可能 BM,1 份(5%)为脑膜炎阴性。可鉴定出肠杆菌科、非发酵菌(铜绿假单胞菌和鲍曼不动杆菌)、溶血葡萄球菌、颗粒杆菌、鞘氨醇单胞菌和粪肠球菌。在脑膜炎患者组中,与培养结果(包括物种鉴定)观察到良好的一致性(4 例中的 3 例)。

结论

分子方法可以补充诊断、指导治疗并识别不可培养的微生物。我们建议对神经外科手术后 BM 的疑似病例联合使用这些方法,特别是在培养结果为阴性的情况下。

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