Rapid detection and simultaneous antibiotic susceptibility analysis of Yersinia pestis directly from clinical specimens by use of reporter phage.
Rapid detection and simultaneous antibiotic susceptibility analysis of Yersinia pestis directly from clinical specimens by use of reporter phage.
复制标题
使用报告噬菌体直接从临床标本中快速检测鼠疫耶尔森氏菌并同时进行抗生素敏感性分析。
DOI：
10.1128/jcm.00316-14
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发表时间：
2014
影响因子：
9.4
通讯作者：
Schofield,DA
中科院分区：
文献类型：
--
作者：
Vandamm,JP;Rajanna,C;Sharp,NJ;Molineux,IJ;Schofield,DA
鼠疫耶尔森氏菌是一个1级代理由于其传染性的肺致病性，非常迅速的进展，和高死亡率。由于这种疾病通常是致命的，没有适当的治疗，从临床矩阵的快速检测是至关重要的病人的结果。我们先前用luxAB工程化诊断噬菌体ΦA1122以产生“光标记”报告噬菌体。ΦA1122：：luxAB快速检测Y。通过转导生物发光信号响应，在纯培养物和人血清中检测鼠疫杆菌。在本报告中，我们评估了报告噬菌体的分析特异性，并直接从加标全血中研究了诊断实用性（检测和抗生素敏感性分析）。生物报告基因对Y的包容性为100%（n= 59）。鼠疫和一致的种内信号转导水平。假阳性没有从典型菌血症相关的物种或鼠疫诊断相关的物种中获得。然而，一些非鼠疫耶尔森氏菌菌株和肠杆菌确实引发信号，尽管在高度衰减的转导水平。在简单肉汤富集血液样本和标准需氧培养瓶中测定诊断性能。在血液中，在5 h内检测到<102 CFU/ml。此外，Y.鼠疫菌可在20至45分钟内从阳性血培养物中直接鉴定出来，无需进一步处理。重要的是，血液样本与抗生素共孵育有助于同时进行抗菌药物敏感性分析。因此，报告噬菌体表现出直接从临床样本中进行快速检测和抗生素敏感性分析，这些特征可以改善鼠疫爆发期间患者的预后。
Yersinia pestis is a tier 1 agent due to its contagious pneumopathogenicity, extremely rapid progression, and high mortality rate. As the disease is usually fatal without appropriate therapy, rapid detection from clinical matrices is critical to patient outcomes. We previously engineered the diagnostic phage ΦA1122 withluxABto create a “light-tagged” reporter phage. ΦA1122::luxABrapidly detects Y. pestis in pure culture and human serum by transducing a bioluminescent signal response. In this report, we assessed the analytical specificity of the reporter phage and investigated diagnostic utility (detection and antibiotic susceptibility analysis) directly from spiked whole blood. The bioreporter displayed 100% (n= 59) inclusivity for Y. pestis and consistent intraspecific signal transduction levels. False positives were not obtained from species typically associated with bacteremia or those relevant to plague diagnosis. However, some non-pestis Yersinia strains and Enterobacteriaceae did elicit signals, albeit at highly attenuated transduction levels. Diagnostic performance was assayed in simple broth-enriched blood samples and standard aerobic culture bottles. In blood, <102CFU/ml was detected within 5 h. In addition, Y. pestis was identified directly from positive blood cultures within 20 to 45 min without further processing. Importantly, coincubation of blood samples with antibiotics facilitated simultaneous antimicrobial susceptibility profiling. Consequently, the reporter phage demonstrated rapid detection and antibiotic susceptibility profiling directly from clinical samples, features that may improve patient prognosis during plague outbreaks.