Agrawal Shailaja, Kulabhusan Prabir Kumar, Joshi Manali, Bodas Dhananjay, Paknikar Kishore M
Nanobioscience group, Agharkar Research Institute, GG Agarkar Road, Pune 411 004, India.
Bioinformatics Centre, Savitribai Phule Pune University, Pune 411007, India.
J Biotechnol. 2016 Aug 10;231:40-45. doi: 10.1016/j.jbiotec.2016.05.027. Epub 2016 May 21.
Salmonellosis is one of the most common and widely distributed foodborne diseases. A sensitive and robust detection method of Salmonella Typhimurium (S. Typhimurium) in food can critically prevent a disease outbreak. In this work, the use of phage displayed peptides was explored for the detection of S. Typhimurium. A phage-displayed random dodecapeptide library was subjected to biopanning against lipopolysaccharide (LPS) of S. Typhimurium. The peptide NFMESLPRLGMH (pep49) derived from biopanning displayed a high affinity (25.8nM) for the LPS of S. Typhimurium and low cross-reactivity with other strains of Salmonella and related Gram-negative bacteria. Molecular insights into the interaction of pep49 with the LPS of S. Typhimurium was gleaned using atomistic molecular dynamics simulations and docking. It was deduced that the specificity of pep49 with S. Typhimurium LPS originated from the interactions of pep49 with abequose that is found only in the O-antigen of S. Typhimurium. Further, pep49 was able to detect S. Typhimurium at a LOD of 10(3) CFU/mL using ELISA, and may be a potential cost efficient alternative to antibodies.
沙门氏菌病是最常见且分布广泛的食源性疾病之一。一种灵敏且可靠的食品中鼠伤寒沙门氏菌（S. Typhimurium）检测方法对于预防疾病爆发至关重要。在这项工作中，探索了利用噬菌体展示肽来检测鼠伤寒沙门氏菌。对一个噬菌体展示的随机十二肽文库进行了针对鼠伤寒沙门氏菌脂多糖（LPS）的生物淘选。从生物淘选中获得的肽NFMESLPRLGMH（pep49）对鼠伤寒沙门氏菌的LPS表现出高亲和力（25.8nM），并且与其他沙门氏菌菌株及相关革兰氏阴性菌的交叉反应性较低。利用原子分子动力学模拟和对接对pep49与鼠伤寒沙门氏菌LPS的相互作用进行了分子层面的深入了解。据推断，pep49与鼠伤寒沙门氏菌LPS的特异性源于pep49与仅在鼠伤寒沙门氏菌O抗原中发现的阿比可糖的相互作用。此外，pep49能够使用酶联免疫吸附测定（ELISA）以10(3) CFU/mL的检测限检测鼠伤寒沙门氏菌，并且可能是一种具有潜在成本效益的抗体替代物。
