A novel artificial Zinc finger – luciferase fusion protein was successfully developed for rapid detection of Salmonella typhimurium, a worldwide-distributed foodborne pathogen. The designed Zinc finger (ZF) protein bound specifically to a 12 bp region of the Salmonella spp invasion gene invA. While the luciferase from Gaussia princeps called Gaussia luciferase (Gluc) was for the first time fused with the artificial ZF domain to improve the detection sensitivity. The fusion protein successfully recognized and bound to the synthesized invA dsDNA with high specificity and sensitivity. The detection limit was as low as 10 fmol of dsNDA. Then, the bacteria PCR products were subsequently used to assess the zinc finger – luciferase fusion protein. The final results indicated that the ZF-Gluc fusion protein system could detect S. typhimurium as low as 1 CFU/mL in 2 h after the PCR. Therefore, this study provided us with a novel artificial zinc finger fusion protein and an efficient method to accomplish the rapid detection of the major foodborne pathogen S. typhimurium. In addition, the specific artificial ZF proteins that bund to particular dsDNA sequences could be easily designed, the ZF-Gluc might has broad application prospects in the field of rapid pathogenic bacteria detection.

成功开发了一种新型人工锌指-荧光素酶融合蛋白，用于快速检测鼠伤寒沙门氏菌，这是一种全球分布的食源性病原体。设计的锌指 (ZF) 蛋白与沙门氏菌属入侵基因invA的 12 bp 区域特异性结合。而来自Gaussiaprinceps的荧光素酶称为Gaussia荧光素酶（Gluc），首次与人工ZF结构域融合，提高了检测灵敏度。融合蛋白成功识别并与合成的invA结合dsDNA 具有高特异性和敏感性。dsNDA 的检测限低至 10 fmol。然后，细菌 PCR 产物随后用于评估锌指 - 荧光素酶融合蛋白。最终结果表明，ZF-Gluc 融合蛋白系统可以在 PCR 后 2 小时内检测低至 1 CFU/mL 的鼠伤寒沙门氏菌。因此，本研究为我们提供了一种新型的人工锌指融合蛋白和一种实现主要食源性病原体鼠伤寒沙门氏菌快速检测的有效方法。此外，与特定dsDNA序列结合的特异性人工ZF蛋白可以很容易地设计，ZF-Gluc在快速病原菌检测领域可能具有广阔的应用前景。