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一种使用逆转录聚合酶链置换反应(RT-PSR)和羟基萘酚蓝指示剂检测活菌的视觉检测方法的开发。

Development of a Visual Assay for Detection of Viable Using RT-PSR and Hydroxynaphthol Blue Indicator.

作者信息

Wang Peng, Chen Qiming, Wang Yikai, Sun Xueting, Liu Zhanmin

机构信息

School of Life Sciences, Shanghai University, No. 99 Shangda Road, Shanghai 200444, China.

Nantong Customs of the People's Republic of China, No. 102 Chongchuan Road, Nantong 226006, China.

出版信息

Biology (Basel). 2025 Apr 7;14(4):383. doi: 10.3390/biology14040383.

DOI:10.3390/biology14040383
PMID:40282248
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12024772/
Abstract

is a foodborne pathogen in powdered infant formula, which poses a significant risk to susceptible populations such as infants and the elderly. This study aims to develop a visual detection method for viable using the reverse transcription-polymerase spiral reaction and hydroxynaphthol blue indicator. Under the optimized conditions, the detection process could be completed within 55 min with low equipment dependence. It was evaluated to have high specificity and sensitivity with the detection limit low to 1.2 × 10 CFU/mL. The assay also showed 100% accuracy in artificially contaminated samples.

摘要

是婴儿配方奶粉中的一种食源性病原体,对婴儿和老年人等易感人群构成重大风险。本研究旨在利用逆转录-聚合酶螺旋反应和羟基萘酚蓝指示剂开发一种用于活菌的视觉检测方法。在优化条件下,检测过程可在55分钟内完成,对设备依赖性低。经评估,该方法具有高特异性和高灵敏度,检测限低至1.2×10 CFU/mL。该检测方法在人工污染样本中也显示出100%的准确率。 (原文中“1.2 × 10 CFU/mL”表述有误,推测可能是“1.2 × 10⁴ CFU/mL”之类,可根据实际情况修正)

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/42a1/12024772/f1d1fbf3c0e9/biology-14-00383-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/42a1/12024772/3b98df5799fc/biology-14-00383-sch001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/42a1/12024772/f1d1fbf3c0e9/biology-14-00383-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/42a1/12024772/3b98df5799fc/biology-14-00383-sch001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/42a1/12024772/f1d1fbf3c0e9/biology-14-00383-g001.jpg

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Infect Immun. 2022 Dec 15;90(12):e0028122. doi: 10.1128/iai.00281-22. Epub 2022 Nov 15.
2
Quantitative Detection of Viable but Nonculturable Using Photosensitive Nucleic Acid Dye PMA Combined with Isothermal Amplification LAMP in Raw Milk.利用光敏核酸染料PMA结合等温扩增技术LAMP对生鲜乳中活的非可培养菌进行定量检测
Foods. 2022 Sep 1;11(17):2653. doi: 10.3390/foods11172653.
3
A simple and sensitive aptasensor with rolling circle amplification for viable Cronobacter sakazakii detection in powdered infant formula.
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4
Quantitative detection of trace VBNC Cronobacter sakazakii by immunomagnetic separation in combination with PMAxx-ddPCR in dairy products.免疫磁珠分离结合 PMAxx-ddPCR 定量检测乳制品中痕量阪崎克罗诺杆菌。
Food Microbiol. 2021 Oct;99:103831. doi: 10.1016/j.fm.2021.103831. Epub 2021 May 19.
5
Trending biocontrol strategies against Cronobacter sakazakii: A recent updated review.针对阪崎克罗诺杆菌的新兴生物防治策略:近期更新综述
Food Res Int. 2020 Nov;137:109385. doi: 10.1016/j.foodres.2020.109385. Epub 2020 Jun 5.
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Gaps in the assortment of rapid assays for microorganisms of interest to the dairy industry.乳制品行业关注的微生物快速检测方法种类存在差距。
Adv Appl Microbiol. 2020;113:1-56. doi: 10.1016/bs.aambs.2020.07.001. Epub 2020 Aug 14.
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A signal cascade amplification strategy based on RT-PCR triggering of a G-quadruplex DNAzyme for a novel electrochemical detection of viable Cronobacter sakazakii.基于 RT-PCR 触发 G-四链体 DNA zyme 的信号级联放大策略,用于新型电活性活菌阪崎克罗诺杆菌检测。
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