Balasubramanian Shankar, Sorokulova Iryna B, Vodyanoy Vitaly J, Simonian Aleksandr L
Materials Research and Education Center, Samuel Ginn College of Engineering, Auburn University, 275 Wilmore, Auburn, AL 36849, USA.
Biosens Bioelectron. 2007 Jan 15;22(6):948-55. doi: 10.1016/j.bios.2006.04.003. Epub 2006 May 12.
Rapid and reliable detection of harmful pathogens at low levels are vital due to the related environmental and economical impact. While antibodies (monoclonal or polyclonal) are successfully employed in many immunoanalysis procedures as a biorecognition element, many of them remain costly with a comparatively short shelf life and uncertain manufacturability. Additionally, they suffer from several limitations, such as susceptibility to hostile environmental stresses such as temperature, pH, ionic strength, and cross-reactivity. The development of easy available, sensitive, and robust alternative molecular recognition elements, capable of providing a very high level of selectivity are very attractive to industry and may benefit in multiple areas. Several attempts have been made to utilize fluorescent-tagged bacteriophages and phage-displayed peptides for bacterial detection. However, involvement of complex labeling and detecting procedures make these approaches time-consuming and complicated. Here, we are reporting for the first time, the label-free detection of Staphylococcus aureus using lytic phage as highly specific and selective biorecognition element and surface plasmon resonance-based SPREETA sensor as a detection platform. Lytic phage was immobilized on the gold surface of SPREETA sensor via trouble-free direct physical adsorption. The detection limit was found to be 10(4) cfu/ml. Detection specificity was investigated by an inhibition assay while selectivity was examined with Salmonella typhimurium. The preliminary results using lytic phage as a probe for bacterial detection, in combination with SPR platform are promising and hence can be employed for rapid and label-free detection of different bacterial pathogens.
由于相关的环境和经济影响，快速可靠地检测低水平的有害病原体至关重要。虽然抗体（单克隆或多克隆）作为生物识别元件已成功应用于许多免疫分析程序中，但其中许多抗体成本高昂，保质期相对较短且可制造性不确定。此外，它们还存在一些局限性，例如易受温度、pH值、离子强度等恶劣环境压力的影响以及交叉反应性。开发易于获得、灵敏且稳健的替代分子识别元件，能够提供非常高的选择性，对工业界极具吸引力，并且可能在多个领域受益。已经进行了几次尝试，利用荧光标记的噬菌体和噬菌体展示肽进行细菌检测。然而，复杂的标记和检测程序使得这些方法耗时且复杂。在此，我们首次报道了使用裂解性噬菌体作为高度特异性和选择性的生物识别元件，以及基于表面等离子体共振的SPREETA传感器作为检测平台，对金黄色葡萄球菌进行无标记检测。裂解性噬菌体通过简便的直接物理吸附固定在SPREETA传感器的金表面。检测限为10(4) cfu/ml。通过抑制试验研究检测特异性，同时用鼠伤寒沙门氏菌检测选择性。使用裂解性噬菌体作为细菌检测探针并结合SPR平台的初步结果很有前景，因此可用于快速无标记检测不同的细菌病原体。
