Ou Aifen, Wang Kan, Mao Yanxiong, Yuan Lei, Ye Yanrui, Chen Ling, Zou Yimin, Huang Tengyi
Department of Food, Guangzhou City Polytechnic, Guangzhou, China.
Center for Translational Medicine, The Second Affiliated Hospital of Shantou University Medical College, Shantou, China.
Front Microbiol. 2021 Jan 7;11:615875. doi: 10.3389/fmicb.2020.615875. eCollection 2020.
Formation of viable but non-culturable (VBNC) status in methicillin-resistant (MRSA) has never been reported, and it poses a significant concern for food safety. Thus, this study aimed to firstly develop a rapid, cost-effective, and efficient testing method to detect and differentiate MRSA strains in the VBNC state and further apply this in real food samples. Two targets were selected for detection of MRSA and toxin, and rapid isothermal amplification detection assays were developed based on cross-priming amplification methodology. VBNC formation was performed for MRSA strain in both pure culture and in artificially contaminated samples, then propidium monoazide (PMA) treatment was further conducted. Development, optimization, and evaluation of PMA-crossing priming amplification (CPA) were further performed on detection of MRSA in the VBNC state. Finally, application of PMA-CPA was further applied for detection on MRSA in the VBNC state in contaminated food samples. As concluded in this study, formation of the VBNC state in MRSA strains has been verified, then two PMA-CPA assays have been developed and applied to detect MRSA in the VBNC state from pure culture and food samples.
耐甲氧西林金黄色葡萄球菌（MRSA）中活的非可培养（VBNC）状态的形成从未被报道过，这对食品安全构成了重大担忧。因此，本研究旨在首先开发一种快速、经济高效的检测方法，以检测和区分处于VBNC状态的MRSA菌株，并进一步将其应用于实际食品样本中。选择了两个靶标用于检测MRSA和毒素，并基于交叉引物扩增方法开发了快速等温扩增检测方法。对纯培养物和人工污染样本中的MRSA菌株进行VBNC形成实验，然后进一步进行单叠氮丙锭（PMA）处理。在检测处于VBNC状态的MRSA时，进一步开展了PMA交叉引物扩增（CPA）的方法开发、优化和评估。最后，将PMA-CPA应用于检测受污染食品样本中处于VBNC状态的MRSA。如本研究所得出的结论，已证实MRSA菌株中VBNC状态的形成，然后开发了两种PMA-CPA检测方法，并将其应用于检测纯培养物和食品样本中处于VBNC状态的MRSA。
