结合LAMP-CRISPR/Cas12b与热敏感尿嘧啶DNA糖苷酶实现可消除残留污染的副溶血弧菌快速可视化检测

吴芳; 卢辰; 胡文灏; 郭馨; 陈佳玥; 罗志丹

doi:10.1631/jzus.B2200705

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结合LAMP-CRISPR/Cas12b与热敏感尿嘧啶DNA糖苷酶实现可消除残留污染的副溶血弧菌快速可视化检测

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Correspondence | Updated：2023-08-08

结合LAMP-CRISPR/Cas12b与热敏感尿嘧啶DNA糖苷酶实现可消除残留污染的副溶血弧菌快速可视化检测
Rapid visual detection of Vibrio parahaemolyticus by combining LAMP-CRISPR/Cas12b with heat-labile uracil-DNA glycosylase to eliminate carry-over contamination
浙江大学学报（英文版）（B辑：生物医学和生物技术） 2023年24卷第8期页码：749-754
Affiliations：

1.Jiangsu Key Laboratory of Marine Biological Resources and Environment, Jiangsu Key Laboratory of Marine Pharmaceutical Compound Screening, Jiangsu Ocean University, Lianyungang 222005, China
2.Co-Innovation Center of Jiangsu Marine Bio-industry Technology, Jiangsu Ocean University, Lianyungang 222005, China
3.BestEnzymes Biotech Co., Ltd., Lianyungang 222005, China
Funds：

the Priority Academic Program Development of Jiangsu Higher Education Institutions of China, the Postgraduate Research & Practice Innovation Program of Jiangsu Province(KYCX2021-038);the Lianyungang Science and Technology Program(CG2232)
DOI：10.1631/jzus.B2200705
中图分类号：
CSTR：

吴芳, 卢辰, 胡文灏, 等. 结合LAMP-CRISPR/Cas12b与热敏感尿嘧啶DNA糖苷酶实现可消除残留污染的副溶血弧菌快速可视化检测[J]. 浙江大学学报（英文版）（B辑：生物医学和生物技术）, 2023,24(8):749-754.

Fang WU, Chen LU, Wenhao HU, et al. Rapid visual detection of Vibrio parahaemolyticus by combining LAMP-CRISPR/Cas12b with heat-labile uracil-DNA glycosylase to eliminate carry-over contamination[J]. Journal of Zhejiang University-SCIENCE B(Biomedicine & Biotechnology), 2023,24(8):749-754.
吴芳, 卢辰, 胡文灏, 等. 结合LAMP-CRISPR/Cas12b与热敏感尿嘧啶DNA糖苷酶实现可消除残留污染的副溶血弧菌快速可视化检测[J]. 浙江大学学报（英文版）（B辑：生物医学和生物技术）, 2023,24(8):749-754. DOI： 10.1631/jzus.B2200705.

Fang WU, Chen LU, Wenhao HU, et al. Rapid visual detection of Vibrio parahaemolyticus by combining LAMP-CRISPR/Cas12b with heat-labile uracil-DNA glycosylase to eliminate carry-over contamination[J]. Journal of Zhejiang University-SCIENCE B(Biomedicine & Biotechnology), 2023,24(8):749-754. DOI： 10.1631/jzus.B2200705.

摘要

Abstract

作为海产品中的主要致病菌，副溶血弧菌的快速准确检测对于海水健康养殖和避免副溶血弧菌相关食源性疾病的发生至关重要。本研究结合尿嘧啶DNA糖苷酶（UDG)、环介导等温扩增（LAMP)和CRISPR/Cas12b技术，建立了海产品中副溶血弧菌的一锅法检测技术。该检测系统实现了一管化检测，并可避免残留污染的风险。我们通过优化dNTP混合物中dTTP/dUTP比例，并筛选出最优sgRNA。该方法在最优条件下，对副溶血弧菌纯培养物的检出限低至1×10,2, CFU/mL，在虾肉样品的检出限低至1×10,2, CFU/g。该方法对其他微生物病原体无交叉反应，且与荧光定量PCR结果符合率为100%。

关键词

尿嘧啶糖苷酶环介导等温扩增CRISPR/Cas12b副溶血弧菌一锅法检测关

Keywords

references

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