Engineering Phage Tail Fiber Protein as a Wide-Spectrum Probe for Acinetobacter baumannii Strains with a Recognition Rate of 100%,Analytical Chemistry

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Engineering Phage Tail Fiber Protein as a Wide-Spectrum Probe for Acinetobacter baumannii Strains with a Recognition Rate of 100%
Analytical Chemistry ( IF 6.7 ) Pub Date : 2022-06-24 , DOI: 10.1021/acs.analchem.2c00682
Ying Chen ₁ , Honglin Yang ₁ , Shuai Luo ₁ , Lin Wang ₁ , Shuguang Lu ₂ , Zhifeng Fu ₁

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As a multidrug-resistant pathogen, Acinetobacter baumannii has long been identified as one of the most common nosocomial bacteria. High-performance recognition probes for wide-spectrum detection of A. baumannii are highly desired to achieve efficient diagnosis and timely treatment of infectious diseases induced by this pathogen. An engineering tail fiber protein (ETFP) named as Gp50 encoded by lytic phage Abp9 was expressed in Escherichia coli and identified as a binding protein for A. baumannii. According to the results of genome sequencing of an A. baumannii wild strain and phage-resistant strains, the binding receptor of ETFP Gp50 is inferred to be a lipopolysaccharide distributed on the bacterial surface. The engineering protein did not show lytic activity to A. baumannii, which facilitates the development of reliable diagnosis kits and biosensors with high flexibility and low false-negative rate. The results of specificity study show that ETFP Gp50 is a species-specific binding protein with a recognition rate of 100% for all tested 77 A. baumannii strains, while that of the natural phage Abp9 is only 27.3%. With the engineering protein, a fluorescence method was developed to detect A. baumannii with a detection range of 2.0 × 10² to 2.0 × 10⁸ cfu mL^–1. The method has been used for the quantification of A. baumannii in a diverse sample matrix with acceptable reliability. The work demonstrates the application potential of ETFP Gp50 as an ideal recognition probe for rapid screening of A. baumannii strains in a complicated sample matrix.

中文翻译：

工程噬菌体尾纤维蛋白作为鲍曼不动杆菌菌株的广谱探针，识别率为100%

作为一种多重耐药病原体，鲍曼不动杆菌长期以来一直被确定为最常见的医院细菌之一。用于广谱检测鲍曼不动杆菌的高性能识别探针对于实现对由该病原体引起的传染病的有效诊断和及时治疗是非常必要的。一种名为 Gp50 的工程尾纤维蛋白 (ETFP) 由裂解噬菌体 Abp9 编码，在大肠杆菌中表达并被鉴定为鲍曼不动杆菌的结合蛋白。根据鲍曼不动杆菌的基因组测序结果野生菌株和噬菌体抗性菌株，ETFP Gp50的结合受体推测为分布于细菌表面的脂多糖。该工程蛋白对鲍曼不动杆菌没有溶解活性，有利于开发具有高灵活性和低假阴性率的可靠诊断试剂盒和生物传感器。特异性研究结果表明，ETFP Gp50是种特异性结合蛋白，对所有测试的77株鲍曼不动杆菌的识别率均为100% ，而天然噬菌体Abp9的识别率仅为27.3%。利用工程蛋白，开发了一种荧光法检测鲍曼不动杆菌，检测范围为2.0 × 10 ²至2.0 × 10 ⁸ cfu mL^–1。该方法已用于对不同样品基质中的鲍曼不动杆菌进行定量，具有可接受的可靠性。该工作证明了 ETFP Gp50 作为一种理想的识别探针在复杂样品基质中快速筛选鲍曼不动杆菌菌株的应用潜力。

更新日期：2022-06-24

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