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首页> 外文期刊>British journal of ophthalmology >Utility of broad-range 16S rRNA PCR assay versus conventional methods for laboratory diagnosis of bacterial endophthalmitis in a tertiary care hospital
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Utility of broad-range 16S rRNA PCR assay versus conventional methods for laboratory diagnosis of bacterial endophthalmitis in a tertiary care hospital

机译：宽范围的16S rRNA PCR测定与常规方法进行三级护理医院中的细菌眼镜炎实验室诊断方法

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Endophthalmitis, a sight-threatening intraocular infection, can be of postsurgical, post-traumatic or endogenous origin. Laboratory diagnosis-based appropriate therapy can be vision-saving. Conventional culture-based laboratory diagnosis takes time and lacks sensitivity. In this study a broad-range PCR assay was assessed against conventional and automated culture methods in vitreous specimens for accurate microbiological diagnosis.To use broad-range PCR assay targeting 16S ribosomal RNA (rRNA) region of bacteria and to assess its performance vis-à-vis conventional and automated culture methods in the laboratory diagnosis of endophthalmitis.Vitreous specimens from 195 patients with clinically diagnosed endophthalmitis were processed for classical and automated culture methods, antimicrobial sensitivity and broad-range PCR assay targeting 762 bp region of 16S rRNA followed by nucleotide sequencing by Sanger’s method. Causative agents were identified from the nucleotide sequences analysed against the GenBank database, and organisms were identified using the Clinical and Laboratory Standards Institute (CLSI) MM18A guidelines.Bacteria could be detected from 127 (65.13%) of the 195 vitreous specimens by broad-range PCR assay; bacterial isolation was possible from 17 (8.7%) and 60 (30.76%) of these specimens by conventional and automated culture methods, respectively (p<0.0001). PCR assay could detect two uncultured bacterium, and in five cases the bacterial identity could not be determined from NCBI database matching.Broad-range PCR assay could provide definitive microbial diagnosis within 24 hours in significantly more patients (p<0.0001). Some rare organisms could be detected, useful in treatment modalities. Automated culture was significantly more sensitive than conventional culture.
机译：眼镜炎，威胁着威胁性的眼内感染，可以是后勤，创伤后或内源性的。实验室诊断的适当疗法可以是节能的。常规的基于培养的实验室诊断需要时间并缺乏敏感性。在该研究中，在玻璃体标本中评估常规和自动化培养方法的广泛PCR测定，用于准确微生物诊断。使用靶向16S核糖体RNA（RRNA）细菌区域的宽范围PCR测定，并评估其性能Vis-on-in-on-l -VIS常规和自动化培养方法在眼镜炎的实验室诊断中。195例临床诊断的患者的患者进行了临床诊断的患者，用于典型和自动化培养方法，抗微生物敏感性和宽范围PCR测定靶向16S rRNA的762bp区域，然后核苷酸靶向核苷酸Sanger方法测序。从针对Genbank数据库分析的核苷酸序列中鉴定了致病剂，并且使用临床和实验室标准研究所（CLSI）MM18A指南来确定生物。通过广泛的玻璃体标本可检测到195个（65.13％）的127（65.13％） PCR测定;通过常规和自动化培养方法分别从17（8.7％）和60（30.76％）的细菌分离可以通过常规和自动化的培养方法（P <0.0001）。 PCR测定可以检测到两个未培养的细菌，并且在五种情况下，不能从NCBI数据库匹配确定细菌标识。BROOD范围PCR测定可以在24小时内提供明确的微生物诊断，在24小时内明显更多的患者（P <0.0001）。可以检测一些稀有生物，可用于治疗方式。自动化文化明显比传统培养更敏感。

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