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实时聚合酶链反应检测引起脑膜炎的最常见细菌和病毒

Real-time PCR Detection of the Most Common Bacteria and Viruses Causing Meningitis.

作者信息

Akkaya Oya, Guvenc Hulya Iren, Yuksekkaya Serife, Opus Aysegul, Guzelant Asuman, Kaya Meral, Kurtoglu Muhammet Guzel, Kaya Nurettin

出版信息

Clin Lab. 2017 Apr 1;63(4):827-832. doi: 10.7754/Clin.Lab.2016.160912.

Abstract

BACKGROUND

Central nervous system (CNS) infections require prompt diagnosis, as the clinical condition progresses rapidly and may lead to severe permanent sequelae or death. The causative agents include viruses, bacteria, fungi, and parasites. In this study, samples with the diagnosis of CNS infection based on cerebrospinal fluid (CSF) sent to us from other hospitals/labs, were studied by multiplex real-time Polymerase Chain Reaction (PCR) method. The purpose of this study is to demonstrate, retrospectively, the most common bacteria and viruses causing meningitis and seasonal distribution of these agents using the multiplex real-time PCR method in CSF samples.

METHODS

This study retrospectively evaluated the results of 470 CSF specimens that had been sent to the Molecular Unit of our hospital with a pre-diagnosis of CNS infection and had been tested with the PCR method between January 2014 and December 2015. Specimens were tested using multiplex real-time PCR assay for Adenovirus (AdV), Cytomegalovirus (CMV), Enteroviruses (EV) (Polioviruses, Coxackieviruses, Echoviruses, and other enteroviruses), Epstein- Barr virus (EBV), Herpes simplex virus 1 and 2, Human Herpes virus 6 and 7, Varicella-zoster virus (VZV), Human Parechoviruses and Parvovirus B19, Hemophilus influenzae, Streptococcus pneumoniae or Neisseria meningitidis. (FTD NEURO9 and FTD Bacterial meningitis, multiplex real-time PCR Kit).

RESULTS

A bacterial or viral agent was identified in 98 (21%) of the 470 CSF samples. Of the patients, 85% were children and 15% were adults. Of the 98 positive samples, 22 (22.5%) patients were 15 years or older, and the remaining 76 (77.5%) were younger than 15 years. While Enterovirus (25%) was the most frequently identified agent, Adenovirus ranked second (22%) and Streptococcus pneumoniae ranked third (15%) in total. Positivity was highest in the 0 - 5-year age range. Bacteria were detected with the PCR method in 22 patients: S. pneumonia in 14, and N. meningitidis in 8. In cultures, S. pneumonia grew only in 7 and N. meningitidis in one. EV and AdV were seen in the summer months. The two coexisted in 3 (3%) patients.

CONCLUSIONS

Early diagnosis and treatment of meningitis are very important for reducing its mortality and morbidity. In patients with suspected meningitis, early detection of the responsible agents may be possible with molecular methods, such as PCR. Significant economic benefits may be obtained by preventing unnecessary antibiotic use and hospitalizations through the early detection of the microbial agents.

摘要

背景

中枢神经系统(CNS)感染需要迅速诊断,因为临床病情进展迅速,可能导致严重的永久性后遗症或死亡。病原体包括病毒、细菌、真菌和寄生虫。在本研究中,我们采用多重实时聚合酶链反应(PCR)方法,对从其他医院/实验室送来的、基于脑脊液(CSF)诊断为CNS感染的样本进行了研究。本研究的目的是通过多重实时PCR方法,回顾性地证明脑脊液样本中引起脑膜炎的最常见细菌和病毒以及这些病原体的季节分布情况。

方法

本研究回顾性评估了2014年1月至2015年12月期间送至我院分子科、预诊断为CNS感染并已用PCR方法检测的470份脑脊液标本的结果。使用多重实时PCR检测试剂盒对标本进行腺病毒(AdV)、巨细胞病毒(CMV)、肠道病毒(EV)(脊髓灰质炎病毒、柯萨奇病毒、埃可病毒和其他肠道病毒)、爱泼斯坦-巴尔病毒(EBV)、单纯疱疹病毒1型和2型、人类疱疹病毒6型和7型、水痘带状疱疹病毒(VZV)、人细小病毒和细小病毒B19、流感嗜血杆菌、肺炎链球菌或脑膜炎奈瑟菌检测。(FTD NEURO9和FTD细菌性脑膜炎多重实时PCR试剂盒)。

结果

在470份脑脊液样本中,98份(21%)检测到细菌或病毒病原体。患者中,85%为儿童,15%为成人。在98份阳性样本中,22例(22.5%)患者年龄在15岁及以上,其余76例(77.5%)年龄小于15岁。总体而言,肠道病毒(25%)是最常检测到的病原体,腺病毒排第二(22%),肺炎链球菌排第三(15%)。0至5岁年龄组的阳性率最高。通过PCR方法在22例患者中检测到细菌:14例为肺炎链球菌,8例为脑膜炎奈瑟菌。在培养物中,仅7例培养出肺炎链球菌,1例培养出脑膜炎奈瑟菌。肠道病毒和腺病毒在夏季出现。3例(3%)患者同时检测到这两种病毒。

结论

脑膜炎的早期诊断和治疗对于降低其死亡率和发病率非常重要。对于疑似脑膜炎患者,通过PCR等分子方法可能早期检测出病原体。通过早期检测微生物病原体,避免不必要的抗生素使用和住院治疗,可获得显著的经济效益。

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