Plant Genetic Engineering Laboratory, School of Agriculture and Food Sciences, The University of Queensland, St. Lucia, Qld, Australia.
Queensland Alliance for Agriculture and Food Innovation, The University of Queensland, St. Lucia, Qld, Australia.
J Appl Microbiol. 2020 Mar;128(3):620-629. doi: 10.1111/jam.14509. Epub 2019 Dec 4.
Current culture-based methods for detection and determination of Campylobacter levels on processed chickens takes at least 2 days. Here we sought to develop a new complete, low-cost and rapid (approximately 2·5 h) detection system requiring minimal operator input.
We observed a strong correlation between culture-based cell counts and our ability to detect either Campylobacter jejuni or Campylobacter coli by loop-mediated isothermal amplification from the same samples. This knowledge was used to develop a rapid and simple five-step assay to quantify Campylobacter, which was subsequently assessed for its specificity, reproducibility and accuracy in quantifying Campylobacter levels from processed chickens. The assay was found to be highly specific for C. jejuni and C. coli and was capable of distinguishing between samples that are either within or exceeding the industry set target of 6000 Campylobacter colony forming units (CFU) per carcass (equivalent to 12 CFU per ml of chicken rinse) with >90% accuracy relative to culture-based methods.
Our method can reliably quantify Campylobacter counts of processed chickens with an accuracy comparable to culture-based assays but provides results within hours as opposed to days.
The research presented here will help improve food safety by providing fast Campylobacter detection that will enable the implementation of real-time risk management strategies in poultry processing plants to rapidly test processed chickens and identify effective intervention strategies. This technology is a powerful tool that can be easily adapted for other organisms and thus could be highly beneficial for a broad range of industries.
目前用于检测和确定加工鸡肉中弯曲杆菌水平的基于培养的方法至少需要 2 天。在这里，我们试图开发一种新的完整、低成本且快速（约 2.5 小时）的检测系统，该系统需要的操作人员输入最少。
我们观察到，基于培养的细胞计数与我们通过环介导等温扩增从相同样本中检测空肠弯曲杆菌或大肠弯曲杆菌的能力之间存在很强的相关性。这一知识被用于开发一种快速而简单的五步检测法来定量弯曲杆菌，随后评估其在定量加工鸡肉中弯曲杆菌水平方面的特异性、重现性和准确性。该检测法对空肠弯曲杆菌和大肠弯曲杆菌具有高度特异性，能够区分低于或超过行业设定的 6000 个弯曲杆菌菌落形成单位（CFU）/胴体目标（相当于每毫升鸡肉冲洗液 12 CFU）的样本，其准确性相对于基于培养的方法>90%。
我们的方法可以可靠地定量加工鸡肉中的弯曲杆菌计数，其准确性可与基于培养的方法相媲美，但在数小时内而不是数天内提供结果。
本文所介绍的研究将通过提供快速的弯曲杆菌检测来帮助提高食品安全，从而使禽肉加工厂能够实时实施风险管理策略，快速测试加工鸡肉并识别有效的干预策略。这项技术是一种强大的工具，可以很容易地适应其他生物体，因此可能对广泛的行业非常有益。
