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通过聚合酶链反应(PCR)从其他假单胞菌属物种中快速、特异性地检测铜绿假单胞菌。

Fast and specific detection of Pseudomonas Aeruginosa from other pseudomonas species by PCR.

作者信息

Jami Al-Ahmadi G, Zahmatkesh Roodsari R

机构信息

Department of Biology, Tonekabon Branch, Islamic Azad University, Iran.

出版信息

Ann Burns Fire Disasters. 2016 Dec 31;29(4):264-267.

Abstract

Pseudomonas aeruginosa is an important life-threatening nosocomial pathogen that plays a prominent role in wound infections of burned patients. We designed this study to identify the isolates of P. aeruginosa recovered from burned patients at the genus and species level through primers targeting oprI and oprL genes, and analyzed their antimicrobial resistance pattern. Over a 2-month period, wound samples were taken from burned patients and plated on MacConkey agar. All suspected colonies were primarily screened for P. aeruginosa by a combination of phenotypic tests. Molecular identifications of colonies were done using specific primers for oprI and oprL genes. Bacterial isolates were recovered from burn wound infections. Based on phenotypical identification tests, 138 (34%) P. aeruginosa isolates were identified; whereas by molecular techniques, just 128 P. aeruginosa yielded amplicon of oprL gene using species-specific primers, verifying the identity of P. aeruginosa; the others yielded amplicon of oprI gene using genus-specific primers, confirming the identity of fluorescent pseudomonads. This study indicates that molecular detection of P. aeruginosa in burn patients employing the OprL gene target is a useful technique for the early and precise detection of P. aeruginosa. PCR detection should be carried out as well as phenotypic testing for the best aggressive antibiotic treatment of P. aeruginosa strains at an earlier stage. It also has significant benefits on clinical outcomes.

摘要

铜绿假单胞菌是一种重要的可危及生命的医院病原体,在烧伤患者的伤口感染中起显著作用。我们设计了本研究,通过靶向oprI和oprL基因的引物,在属和种水平上鉴定从烧伤患者中分离出的铜绿假单胞菌,并分析其抗菌耐药模式。在2个月的时间里,从烧伤患者身上采集伤口样本,接种在麦康凯琼脂平板上。所有疑似菌落首先通过表型试验组合进行铜绿假单胞菌的初步筛查。使用针对oprI和oprL基因的特异性引物对菌落进行分子鉴定。从烧伤伤口感染中分离出细菌菌株。基于表型鉴定试验,鉴定出138株(34%)铜绿假单胞菌分离株;而通过分子技术,只有128株铜绿假单胞菌使用种特异性引物产生了oprL基因扩增子,证实了铜绿假单胞菌的身份;其他菌株使用属特异性引物产生了oprI基因扩增子,证实了荧光假单胞菌的身份。本研究表明,采用OprL基因靶点对烧伤患者中的铜绿假单胞菌进行分子检测是一种用于早期和精确检测铜绿假单胞菌的有用技术。为了在早期对铜绿假单胞菌菌株进行最佳的积极抗生素治疗,应同时进行PCR检测和表型检测。这对临床结果也有显著益处。

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