Zhong Junliang, Zhao Xihong
Key Laboratory for Green Chemical Process of Ministry of Education, Key Laboratory for Hubei Novel Reactor and Green Chemical Technology, Research Center for Environmental Ecology and Engineering, School of Environmental Ecology and Biological Engineering, Wuhan Institute of Technology, Wuhan, 430205 China.
3 Biotech. 2018 Jan;8(1):28. doi: 10.1007/s13205-017-1052-7. Epub 2017 Dec 22.
The aim of this study was to evaluate the applicability of the conventional PCR detection method combined with propidium monoazide (PMA) treatment for the detection of viable but non-culturable (VBNC) state O157:H7 in ground beef meatballs. Under low temperature, O157:H7 cells were induced into the VBNC state in ground beef meatballs at - 20 °C after 152 days. The optimal PMA concentration of 5 µg/mL was obtained in beef meatball samples, which could completely inhibit the DNA amplification on dead cells (10 cells/mL) but with no inhibition on viable cells. The established PMA-PCR assay revealed that the VBNC counts exceeded 10 CFU/mL in artificial contamination beef samples, which could be used for semi-quantitative detection of VBNC cells in beef meatball samples. This study indicated that the PMA-PCR assay might be a potential method for detection of VBNC state . O157:H7 cells in food products.
本研究的目的是评估常规PCR检测方法结合单叠氮化丙锭(PMA)处理用于检测牛肉丸中活的但不可培养(VBNC)状态的O157:H7的适用性。在低温下,152天后,O157:H7细胞在-20°C的牛肉丸中被诱导进入VBNC状态。在牛肉丸样品中获得了5μg/mL的最佳PMA浓度,该浓度可完全抑制死细胞(10个细胞/mL)的DNA扩增,但对活细胞无抑制作用。所建立的PMA-PCR检测方法表明,人工污染牛肉样品中VBNC数量超过10 CFU/mL,可用于半定量检测牛肉丸样品中的VBNC细胞。本研究表明,PMA-PCR检测方法可能是检测食品中VBNC状态的O157:H7细胞的一种潜在方法。