Cranfield University, Cranfield, Bedfordshire, MK43 0AL, England, United Kingdom.
Biotechnology and Nanotechnology Research Center, Malaysian Agricultural Research and Development Institute, MARDI, P. O. Box 12301, 50774, Kuala Lumpur, Malaysia.
Sci Rep. 2019 Sep 20;9(1):13642. doi: 10.1038/s41598-019-49672-2.
In this work, a subtractive inhibition assay (SIA) based on surface plasmon resonance (SPR) for the rapid detection of Campylobacter jejuni was developed. For this, rabbit polyclonal antibody with specificity to C. jejuni was first mixed with C. jejuni cells and unbound antibody was subsequently separated using a sequential process of centrifugation and then detected using an immobilized goat anti-rabbit IgG polyclonal antibody on the SPR sensor chip. This SIA-SPR method showed excellent sensitivity for C. jejuni with a limit of detection (LOD) of 131 ± 4 CFU mL and a 95% confidence interval from 122 to 140 CFU mL. The method has also high specificity. The developed method showed low cross-reactivity to bacterial pathogens such as Salmonella enterica serovar Typhimurium (7.8%), Listeria monocytogenes (3.88%) and Escherichia coli (1.56%). The SIA-SPR method together with the culturing (plating) method was able to detect C. jejuni in the real chicken sample at less than 500 CFU mL, the minimum infectious dose for C. jejuni while a commercial ELISA kit was unable to detect the bacterium. Since the currently available detection tools rely on culturing methods, which take more than 48 hours to detect the bacterium, the developed method in this work has the potential to be a rapid and sensitive detection method for C. jejuni.
在这项工作中,开发了一种基于表面等离子体共振(SPR)的减法抑制检测(SIA)方法,用于快速检测空肠弯曲菌。为此,首先将针对空肠弯曲菌的兔多克隆抗体与空肠弯曲菌细胞混合,然后使用离心的连续过程分离未结合的抗体,并用 SPR 传感器芯片上固定的兔抗羊 IgG 多克隆抗体进行检测。这种 SIA-SPR 方法对空肠弯曲菌具有优异的灵敏度,检测限(LOD)为 131±4 CFU mL,95%置信区间为 122 至 140 CFU mL。该方法还具有高度特异性。开发的方法对沙门氏菌血清型 Typhimurium(7.8%)、李斯特菌(3.88%)和大肠杆菌(1.56%)等细菌病原体的交叉反应性较低。SIA-SPR 方法与培养(平板)方法相结合,能够在低于 500 CFU mL 的浓度下检测真实鸡肉样本中的空肠弯曲菌,这是空肠弯曲菌的最小感染剂量,而商业 ELISA 试剂盒无法检测到该细菌。由于现有的检测工具依赖于培养方法,需要 48 小时以上才能检测到细菌,因此本工作中开发的方法有可能成为一种快速、灵敏的空肠弯曲菌检测方法。