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分子生物学技术在急性术后眼内炎微生物诊断中的原理及应用。

Principles and applications of molecular biology techniques for the microbiological diagnosis of acute post-operative endophthalmitis.

机构信息

Department of Ophthalmology, Hospices Civils de Lyon, Université Lyon I, Lyon, France.

Department of Microbiology, CHU de Grenoble, Université Joseph Fourier, Grenoble, France; Department of Microbiology, Hospices Civils de Lyon, Université Lyon 1, Centre, National de référence des Staphylocoques, and INSERM, U851, Bron, France.

出版信息

Surv Ophthalmol. 2014 May-Jun;59(3):286-303. doi: 10.1016/j.survophthal.2013.08.002. Epub 2013 Dec 18.

DOI:10.1016/j.survophthal.2013.08.002
PMID:24359808
Abstract

The systematic microbiological evaluation of endophthalmitis allows the confirmation of the infectious nature of the disease and the possible adaptation of treatment at the individual level and, at the collective level, the epidemiological characterization of the bacterial spectrum of endophthalmitis. Long reserved for research, the use of molecular biology techniques to complement conventional culture techniques has become important for the diagnosis of endophthalmitis in recent years. These new diagnostic techniques are particularly useful for the microbiological study of bacteria that are difficult or impossible to grow because of their intrinsic properties, their presence in only a small inoculum, their sequestration on prosthetic materials, or their inactivation by prior antibiotic treatment. These techniques are based on the polymerase chain reaction (PCR), which allows the amplification and detection of extracted bacterial deoxyribonucleic acid (DNA) that is initially present in minute quantities in an ocular sample. In practice, these conventional or real-time PCRs allow either the a priori detection of bacterial DNA (universal PCR) or the identification of a specific DNA fragment of a bacterial genus or species (specific PCR). New techniques of PCR will allow more rapid bacterial identification and also characterization of genotypic properties, such as genes of virulence or antibiotic resistance.

摘要

眼内炎的系统微生物学评估可以确认疾病的感染性质,并可能在个体层面上调整治疗方案,从集体层面上对眼内炎的细菌谱进行流行病学特征描述。分子生物学技术曾经长期用于研究,近年来已成为眼内炎诊断的重要手段,用于补充传统的培养技术。这些新技术对于研究由于内在特性、菌量少、定植于假体材料或先前抗生素治疗而导致难以或无法生长的细菌的微生物学特别有用。这些技术基于聚合酶链反应(PCR),可以扩增和检测从眼部样本中提取的初始含量很少的细菌脱氧核糖核酸(DNA)。实际上,这些常规或实时 PCR 可以预先检测细菌 DNA(通用 PCR)或鉴定细菌属或种的特定 DNA 片段(特异性 PCR)。新的 PCR 技术将使细菌鉴定更快,并能够对毒力或抗生素耐药性等基因型特性进行特征描述。

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