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首页> 外文期刊>Clinical and diagnostic laboratory immunology >Detection of Francisella tularensis in Biological Specimens Using a Capture Enzyme-Linked Immunosorbent Assay, an Immunochromatographic Handheld Assay, and a PCR
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Detection of Francisella tularensis in Biological Specimens Using a Capture Enzyme-Linked Immunosorbent Assay, an Immunochromatographic Handheld Assay, and a PCR

机译：使用捕获酶联免疫吸附测定，免疫色谱手持测定和PCR检测生物标本中的弗拉杆菌

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The early detection of Francisella tularensis, the causative agent of tularemia, is important for adequate treatment by antibiotics and the outcome of the disease. Here we describe a new capture enzyme-linked immunosorbent assay (cELISA) based on monoclonal antibodies specific for lipopolysaccharide (LPS) of Francisella tularensis subsp. holarctica and Francisella tularensis subsp. tularensis. No cross-reactivity with Francisella tularensis subsp. novicida,Francisella philomiragia, and a panel of other possibly related bacteria, including Brucella spp.,Yersinia spp., Escherichia coli, andBurkholderia spp., was observed. The detection limit of the assay was 103 to 104 bacteria/ml. This sensitivity was achieved by solubilization of the LPS prior to the cELISA. In addition, a novel immunochromatographic membrane-based handheld assay (HHA) and a PCR, targeting sequences of the 17-kDa protein (TUL4) gene of F. tularensis, were used in this study. Compared to the cELISA, the sensitivity of the HHA was about 100 times lower and that of the PCR was about 10 times higher. All three techniques were successfully applied to detect F. tularensis in tissue samples of European brown hares (Lepus europaeus). Whereas all infected samples were recognized by the cELISA, those with relatively low bacterial load were partially or not detected by PCR and HHA, probably due to inhibitors or lack of sensitivity. In conclusion, the HHA can be used as a very fast and simple approach to perform field diagnosis to obtain a first hint of an infection with F. tularensis, especially in emergent situations. In any suspect case, the diagnosis should be confirmed by more sensitive techniques, such as the cELISA and PCR.
机译：早期发现图拉弗朗西斯菌是图拉姆病的病原体，对于抗生素的适当治疗和疾病的结果非常重要。在这里，我们描述了一种新的捕获酶联免疫吸附试验（cELISA），该酶基于特异性针对 Francisella tularensis 亚种脂多糖（LPS）的单克隆抗体。 holarctica 和 Francisella tularensis 子亚种。 tularensis 。与 ella Tularensis 亚种无交叉反应。 novicida ， Francisella philomiragia ，以及一组其他可能相关的细菌，包括 Brucella 属，耶尔森氏菌。 ，大肠杆菌和伯克霍尔德菌 spp.。该方法的检出限为10 3 至10 4 / ml。通过在cELISA之前溶解LPS可以达到这种敏感性。此外，新型的基于免疫层析膜的手持分析（HHA）和PCR靶向了 F的17-kDa蛋白（TUL4）基因的序列。在本研究中使用了tularensis 。与cELISA相比，HHA的灵敏度低约100倍，PCR的灵敏度高约10倍。所有这三种技术已成功应用于检测 F。欧洲褐兔（ eurpueus ）组织样品中的土拉木（tularensis）。尽管所有感染的样品均通过cELISA识别，但细菌载量相对较低的样品只能通过抑制剂或缺乏敏感性而通过PCR和HHA部分检测或未检测到。总之，HHA可以用作执行现场诊断的非常快速和简单的方法，以获得F感染的初步提示。 tularensis ，尤其是在紧急情况下。在任何可疑情况下，都应通过更敏感的技术（例如cELISA和PCR）来确认诊断。

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