PMA-qPCR技术在发酵食品活菌计数中的应用

Application of PMA-qPCR in enumerating living microbes in fermented foods

发酵食品具有丰富的口感和较高的营养价值,深受广大消费者喜爱。准确计数发酵食品生产加工与贮藏过程中的活菌,是对产品进行质量控制、评价以及工艺改进的基础。叠氮溴化丙锭(propidium monoazide,PMA)是一种具有高亲和力的光敏性DNA结合染料,用于处理样品后偶联使用实时荧光定量聚合酶链式反应(quantitative real-time PCR,qPCR)计数可极大缩短检测时长且灵敏度高、特异性强。该文综述PMA-qPCR技术的原理、影响因素及其在红酒、啤酒、酸奶等发酵食品活菌计数中的应用现状。将目标微生物及PMA处理条件列为此技术影响因素,以此为基础总结各类发酵食品中可检测微生物及检测线性范围及检测限,为扩大此方法使用范围,获取发酵食品中更为丰富和精准的活菌检测结果提供参考。

Fermented foods are favored by consumers because of their high nutritional values and tastes.Monitoring living microorganisms during processing and storage of fermented foods is important for quality control,production evaluation and process modification.Propidium monoazide(PMA)is a highly photosensitive DNA-binding dye that can be used with quantitative fluorescence real-time PCR(qPCR)for quick,accurate,and specific detection.The mechanisms and influencing factors of PMA-qPCR,as well as its applications in enumerating living microbes in wines,beers,and yoghurts were reviewed.It was found that the types of target microorganisms and PMA treatment conditions were the most important influencing factors.The linear ranges and detection limits of detectable microbes in various fermented foods were summarized.This review aimed to provide a reference for multi-faceted applications of PMA-qPCR.