叠氮溴化丙锭结合实时定量聚合酶链式反应法快速检测玫瑰酱中戊糖片球菌活菌-食品安全质量检测学报-CSCIED科技核心评价数据库-手机版

叠氮溴化丙锭结合实时定量聚合酶链式反应法快速检测玫瑰酱中戊糖片球菌活菌

Rapid detection of viable Pediococcus pentosaceus in rose jam by propidium monoazide-quantitative real-time polymerase chain reaction method

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DOI	10.19812/j.cnki.jfsq11-5956/ts.20250110007
刊名	食品安全质量检测学报 Journal of Food Safety and Quality
年，卷(期)	2025, 16(13)
作者	张艳珍, 袁伟杰, 马超, 麻丽丹, 刘云国*,
作者单位	临沂大学生命科学学院; 水原大学工程学院; 拱北海关技术中心
摘要	目的探究基于叠氮溴化丙锭(propidium monoazide, PMA)与实时定量聚合酶链式反应(quantitative real-time polymerase chain reaction, qPCR)技术联用, 建立一种快速检测玫瑰酱中戊糖片球菌活菌的方法。方法根据特异性基因murI设计戊糖片球菌特异性引物, 通过优化PMA浓度、暗孵育时间和曝光时间, 确定最佳PMA处理条件, 并通过PMA-qPCR方法验证不同比例的戊糖片球菌死菌/活菌混合菌液, 评估该方法的灵敏度和检出限。结果设计的引物具有良好的特异性, PMA的最佳处理条件为质量浓度80 μg/mL, 暗孵育时间为15 min, 曝光时间为25 min, 此条件下, 构建的标准曲线相关系数(r2)为0.9983, 最低检出限为3.98×103 CFU/g。结论本研究建立的PMA-qPCR方法可有效实现玫瑰酱中戊糖片球菌活菌数的快速检测, 灵敏度高, 为复杂食品基质中微生物的定量检测提供了技术支持。
Abstract	Objective To establish a rapid detection method of viable Pediococcus pentosaceus in rose jam based on the combination of propidium monoazide (PMA) and quantitative real-time polymerase chain reaction (qPCR). Methods Primers specific for Pediococcus pentosaceus were designed according to the species-specific murI gene. The optimal PMA treatment conditions were determined by optimizing the PMA concentration, dark incubation time, and exposure time. Different ratios of Pediococcus pentosaceus dead/live bacterial mixtures were validated by the PMA-qPCR method to assess the sensitivity and limit of detection of the technique. Results The designed primers exhibited high specificity, and the optimal PMA treatment conditions were determined as 80 μg/mL PMA, 15 min of dark incubation, and 25 min of exposure. Under this condition, a standard curve with correlation coefficient (r2) of 0.9983 was constructed, and the lowest limit of detection was determined to be 3.98×103 CFU/g. Conclusion The PMA-qPCR method established in this study enables the rapid and accurate quantification of viable Pediococcus pentosaceus in rose jam with high sensitivity, providing technical support for the quantitative detection of microorganisms in complex food matrices.
关键词	玫瑰酱;戊糖片球菌;propidium monoazide;quantitative real-time polymerase chain reaction;活菌检测
KeyWord	rose jam; Pediococcus pentosaceus; propidium monoazide; quantitative real-time polymerase chain reaction; viable
基金项目
页码	214-219

参考文献
相关文献

[1]QI Y, HUANG L, ZENG Y, et al. Pediococcus pentosaceus: screening and application as probiotics in food processing
[J]. Frontiers in Microbiology, 2021, 12: 762467.
[2]JIANG S, CAI L, LV L, et al. Pediococcus pentosaceus, a future additive or probiotic candidate
[J]. Microbial Cell Factories, 2021, 20: 45.
[3]SABOORI B, SHAHIDI F, HEDAYATI S, et al. Investigating the probiotic properties and antimicrobial activity of lactic acid bacteria isolated from an Iranian fermented dairy product, Kashk
[J]. Foods, 2022, 11(23): 3904.
[4]HA JH, KIM AR, LEE KS, et al. Anti-aging activity of Lavandula angustifolia extract fermented with Pediococcus pentosaceus DK1 isolated from Diospyros kaki fruit in UVB-irradiated human skin fibroblasts and analysis of principal components
[J]. Journal of Microbiology and Biotechnology, 2019, 29(1): 21–29.
[5]AHMED S, ASHRAF F, TARIQ M, et al. Aggrandizement of fermented cucumber through the action of autochthonous probiotic cum starter strains of Lactiplantibacillus plantarum and Pediococcus pentosaceus
[J]. Annals of Microbiology, 2021, 71(1): 33.
[6]LU K, WANG X, ZHOU Y, et al. Genomic characterization and probiotic potential assessment of an exopolysaccharide-producing strain Pediococcus pentosaceus LL-07 isolated from fermented meat
[J]. BMC Microbiology, 2024, 24(1): 142.
[7]JEONGHYUN Y, TAE WK, CHANG WC, et al. Antifungal mechanisms investigation of lactic acid bacteria against Aspergillus flavus: Through combining microbial metabolomics and co-culture system
[J]. Journal of Applied Microbiology, 2024, 135(5): 112.
[8]ZHANG Y, HU P, XIE Y, et al. Co-fermentation with Lactobacillus curvatus LAB26 and Pediococcus pentosaceus SWU73571 for improving quality and safety of sour meat
[J]. Meat Science, 2020, 170: 108240.
[9]SONG G, HE Z, WANG X, et al. Improving the quality of Suancai by inoculating with Lactobacillus plantarum and Pediococcus pentosaceus
[J]. Food Research International, 2021, 148: 110581.
[10]FUJIWARA M, KUWAHARA D, HAYASHI M, et al. Lowering effect of viable Pediococcus pentosaceus QU 19 on the rise in postprandial glucose
[J]. Bioscience of Microbiota, Food and Health, 2020, 39(2): 57–64.
[11]CAO Z, PAN H, TONG H, et al. In vitro evaluation of probiotic potential of Pediococcus pentosaceus L1 isolated from paocai-a Chinese fermented vegetable
[J]. Annals of Microbiology, 2016, 66(3): 963–971.
[12]KIM JS, KIM JH, PALANIYANDI SA, et al. Yak-Kong soybean (Glycine max) fermented by a novel Pediococcus pentosaceus inhibits the oxidative stress-induced monocyte–endothelial cell adhesion
[J]. Nutrients, 2019, 11(6): 1380.
[13]BIAN X, YANG L, WU W, et al. Pediococcus pentosaceus LI05 alleviates DSS-induced colitis by modulating immunological profiles, the gut microbiota and short-chain fatty acid levels in a mouse model
[J]. Microbial Biotechnology, 2020, 13(4): 1228–1244.
[14]SHIN JS, JUNG JY, LEE SG, et al. Exopolysaccharide fraction from Pediococcus pentosaceus KFT18 induces immunostimulatory activity in macrophages and immunosuppressed mice
[J]. Journal of Applied Microbiology, 2016, 120(5): 1390–1402.
[15]郑淑彦, 王伟, 董金金, 等. 食用玫瑰营养保健功能及产品开发研究进展
[J]. 食品研究与开发, 2016, 37(23): 206–211. ZHENG SY, WANG W, DONG JJ, et al. Research progress on nutrition health function and product development of edible rose
[J]. Food Research and Development, 2016, 37(23): 206–211.
[16]杨文清, 李梦茹, 李西西, 等. 玫瑰酱的功能特性与香气成分分析
[J]. 中国调味品, 2023, 48(3): 156–160, 169. YANG WQ, LI MR, LI XX, et al. Analysis of functional properties and aroma components of rose sauce
[J]. China Condiment, 2023, 48(3): 156–160, 169.
[17]续钊, 张元馨, 卢前明. 玫瑰酱中微生物多样性分析及优势菌群的鉴定
[J]. 中国调味品, 2023, 48(3): 179–182. XU Z, ZHANG YX, LU QM. Analysis of microbial diversity and identification of dominant microbial communities in rose sauce
[J]. China Condiment, 2023, 48(3): 179–182.
[18]夏傲喃. 玫瑰酱中微生物多样性分析及优势菌群的鉴定
[D]. 乌鲁木齐: 新疆大学, 2021. XIA AON. Analysis of microbial diversity and identification of dominant flora in rose jam
[D]. Urumqi: Xinjiang University, 2021.
[19]陈卓君, 魏铭, 林果, 等. PMA-qPCR技术在发酵食品活菌计数中的应用
[J]. 食品与发酵工业, 2019, 12(45): 242–248. CHEN ZJ, WEI M, LIN G, et al. Application of PMA-qPCR in enumerating living microbes in fermented foods
[J]. Food and Fermentation Industries, 2019, 12(45): 242–248.
[20]王力均, 谭强来, 朱江, 等. 应用PMA-qPCR方法快速准确检测发酵乳制品中副干酪乳杆菌活菌的研究
[J]. 中国微生态学杂志, 2013, 25(1): 1–4. WANG LJ, TAN QL, ZHU J, et al. Development of PMA-qPCR assay for rapid and accurate detection of viable Lactobacillus paracasei in fermented dairy products
[J]. Chinese Journal of Microecology, 2013, 25(1): 1–4.
[21]张亚豪, 梁会朋, 常聪, 等. 基干实时荧光定量PCR技术监测四川工业泡菜发酵过程中主要细菌的变化
[J]. 中国调味品, 2018, 43(1): 35–38, 47. ZHANG YH, LIANG HP, CHANG C, et al. The variation of main bacteria in fermentation of Sichuan industrial pickles monitored by real-time fluorescence quantitative PCR
[J]. China Condiment, 2018, 43(1): 35–38, 47.
[22]盖冬雪, 任洪林, 卢士英, 等. 乳品中大肠杆菌PMA-qPCR活菌检测方法的建立
[J]. 中国畜牧兽医, 2016, 43(2): 493–498. GAI DX, REN HL, LU SY, et al. Establishment of PMA-qPCR assay for detection of viable E.coli in milk
[J]. China Animal Husbandry & Veterinary Medicine, 2016, 43(2): 493–498.
[23]陶怡君. 叠氮溴化丙锭结合qPCR检测与区分活菌和死菌的研究进展
[J]. 微生物学免疫学进展, 2020, 48(6): 63–68. TAO YJ. Advances in detection and differentiation of live and dead bacteria by PMA-qPCR technology
[J]. Progress In Microbiology and Immunology, 2020, 48(6): 63–68.
[24]彭琳媛, 林洪, 王静雪. PMA-qPCR定量检测VBNC副溶血弧菌方法的建立与优化
[J]. 食品与发酵工业, 2022, 48(1): 247–252. PENG LY, LIN H, WANG JX. Development and optimization of PMA-qPCR for the quantitative detection of viable but non-culturable Vibrio parahaemolyticus
[J]. Food and Fermentation Industries, 2022, 48(1): 247–252.
[25]吕旭聪, 刘志彬, 张雯, 等. 红曲黄酒传统酿造过程中优势细菌活菌检测方法的构建及应用
[J]. 中国食品学报, 2018, 18(7): 210–219. LU XC, LIU ZB, ZHANG W, et al. Construction and application of detection method for viable dominant bacteria during traditional brewing of Hong Qu glutinous rice wine
[J]. Journal of Chinese Institute of Food Science and Technology, 2018, 18(7): 210–219.
[26]QAMAR H, HE R, LI Y, et al. Metabolome and metagenome integration unveiled synthesis pathways of novel antioxidant peptides in fermented lignocellulosic biomass of palm kernel meal
[J]. Antioxidants, 2024, 13(10): 1253.
[27]谢鑫, 粱云, 郭立芸. 荧光定量PCR检测啤酒酿造过程中的戊糖片球菌
[J]. 中外酒业, 2016, 4(7): 19–24. XIE X, LIANG Y, GUO LY. Detection of Pediococcus pentosaceus during beer brewing by fluorescence quantitative PCR
[J]. Global Alcinfo, 2016, 4(7): 19–24.
[28]GUO J, FAN F, WANG W, et al. Development of PMA-qPCR assay to accurately and reproducible quantify viable bacteria of Paenibacillus polymyxa
[J]. Letters in Applied Microbiology, 2023, 76(11): ovad127.
[29]曹怡芳, 周爱莲, 白泓, 等. 牛奶发酵过程中阪崎克罗诺杆菌可形成活的非可培养状态
[J]. 现代食品科技, 2022, 38(3): 56–62. CAO YF, ZHOU AIL, BAI H, et al. Formation of viable but non-culturable state of Cronobacter sakazakii during milk fermention
[J]. Modern Food Science and Technology, 2022, 38(3): 56–62.
[30]段亮杰, 沙雨婷, 罗意, 等. 叠氮溴化丙锭-荧光定量PCR法实时快速检测5种乳杆菌活菌数方法的建立与应用
[J]. 微生物学通报, 2020, 47(12): 4317–4327. DUAN LJ, SHA YT, LUO Y, et al. Quantitative PCR combined with propidium monoazide treatment for real-time and rapid determination of five viable Lactobacillus
[J]. Microbiology China, 2020, 47(12): 4317–4327.
[31]李畅, 任洪林, 卢士英, 等. 乳品中的酵母活菌数PMA-qPCR检测方法的建立及初步应用
[J]. 食品工业科技, 2018, 39(22): 128–132. LI C, REN HL, LU SY, et al. Development and application of PMA-qPCR method for detecting viable yeasts in dairy products
[J]. Science and Technology of Food Industry, 2018, 39(22): 128–132.
[32]SHI Z, LI X, FAN X, et al. PMA-qPCR method for the selective quantitation of viable lactic acid bacteria in fermented milk
[J]. Frontiers in Microbiology, 2022, 13: 984506.
[33]WANG X, CHEN C, SHEN T, et al. Heterologous expression, purification and biochemical characterization of a glutamate racemase (MurI) from Streptococcus mutans UA159
[J]. Peer J Life & Environment, 2019, 7: e8300.
[34]马丹, 杨彬彬, 陶文靖, 等. 叠氮溴化丙锭-qPCR法定量检测植物乳杆菌
[J]. 食品安全质量检测学报, 2021, 12(12): 4867–4875. MA D, YANG BB, TAO WJ, et al. Quantitative detection of Lactobacillus plantarum by propidium monoazide-qPCR
[J]. Journal of Food Safety & Quality, 2021, 12(12): 4867–4875.

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张艳珍, 袁伟杰, 马超, 麻丽丹, 刘云国*. 叠氮溴化丙锭结合实时定量聚合酶链式反应法快速检测玫瑰酱中戊糖片球菌活菌 [J]. 食品安全质量检测学报. 2025; 16; (13). 214 - 219.

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