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食品基质对基于toxR基因的水产品中副溶血性弧菌快速检测的影响

Effect of food matrix on rapid detection of Vibrio parahaemolyticus in aquatic products based on toxR gene.

作者信息

Li Darong, Zhao Jiayi, Lan Weiqing, Zhao Yong, Sun Xiaohong

机构信息

College of Food Science and Technology, Shanghai Ocean University, Shanghai, 201306, People's Republic of China.

Shanghai Engineering Research Center of Aquatic-Product Processing & Preservation, Shanghai, 201306, People's Republic of China.

出版信息

World J Microbiol Biotechnol. 2023 May 9;39(7):188. doi: 10.1007/s11274-023-03640-1.

DOI:10.1007/s11274-023-03640-1
PMID:37156898
Abstract

Vibrio parahaemolyticus has become an important public threat to human health. Rapid and robust pathogen diagnostics are necessary for monitoring its outbreak and spreading. Herein, we report an assay for the detection of V. parahaemolyticus based on recombinase aided amplification (RAA) combined with lateral flow dipstick (LFD), namely RAA-LFD. The RAA-LFD took 20 min at 36~38 ℃, and showed excellent specificity. It detected as low as 6.4 fg/µL of V. parahaemolyticus in genomic DNA, or 7.4 CFU/g spiked food samples with 4 h of enrichment. The limit of detection in shrimp (Litopenaeus Vannamei), fish (Carassius auratus), clam (Ruditapes philippinarum) evidenced that sensitivity was considerably affected by the food matrix. The presence of food matrix reduced the sensitivity of spiked food samples by 10 ~ 100 times. In the filed samples detection, RAA-LFD method showed good coincidence with GB4789.7-2013 method and PCR method at rates of 90.6% and 94.1%, respectively. RAA-LFD has high accuracy and sensitivity for the detection of V. parahaemolyticus, which can serve as a model tool to meet the growing need for point-of-care diagnosis of V. parahaemolyticus.

摘要

副溶血性弧菌已成为对人类健康的重要公共威胁。快速且可靠的病原体诊断对于监测其爆发和传播至关重要。在此，我们报告一种基于重组酶辅助扩增（RAA）结合侧向流动试纸条（LFD）检测副溶血性弧菌的方法，即RAA-LFD。RAA-LFD在36~38℃下只需20分钟，且具有出色的特异性。它能检测低至6.4 fg/µL基因组DNA中的副溶血性弧菌，或在经过4小时富集的加标食品样本中检测到低至7.4 CFU/g的副溶血性弧菌。在虾（凡纳滨对虾）、鱼（鲫鱼）、蛤（菲律宾蛤仔）中的检测限表明，食品基质对灵敏度有显著影响。食品基质的存在使加标食品样本的灵敏度降低了10至100倍。在现场样本检测中，RAA-LFD方法与GB4789.7-2013方法和PCR方法的符合率分别为90.6%和94.1%，显示出良好的一致性。RAA-LFD对副溶血性弧菌的检测具有高准确性和灵敏度，可作为满足对副溶血性弧菌即时诊断日益增长需求的模型工具。

相似文献

[1]
Effect of food matrix on rapid detection of Vibrio parahaemolyticus in aquatic products based on toxR gene.

World J Microbiol Biotechnol. 2023-5-9

[2]
Improvement and evaluation of loop-mediated isothermal amplification combined with chromatographic flow dipstick assays for Vibrio parahaemolyticus.

J Microbiol Methods. 2020-4

[3]
Development and evaluation of a sensitive recombinase aided amplification assay for rapid detection of Vibrio parahaemolyticus.

J Microbiol Methods. 2022-2

[4]
Rapid and Visual Detection of in Aquatic Foods Using Gene-Based Loop-Mediated Isothermal Amplification with Lateral Flow Dipstick (LAMP-LFD).

J Microbiol Biotechnol. 2021-12-28

[5]
The development of loop-mediated isothermal amplification combined with lateral flow dipstick for detection of Vibrio parahaemolyticus.

Lett Appl Microbiol. 2011-2-7

[6]
Recombinase polymerase amplification-lateral flow (RPA-LF) assay combined with immunomagnetic separation for rapid visual detection of Vibrio parahaemolyticus in raw oysters.

Anal Bioanal Chem. 2020-5

[7]
Rapid and Sensitive Detection of Vibrio parahaemolyticus and Vibrio vulnificus by Multiple Endonuclease Restriction Real-Time Loop-Mediated Isothermal Amplification Technique.

Molecules. 2016-1-19

[8]
An improved recombinase polymerase amplification assay for visual detection of Vibrio parahaemolyticus with lateral flow strips.

J Food Sci. 2020-5-25

[9]
Development of an isothermal recombinase-aided amplification assay for the rapid and visualized detection of Klebsiella pneumoniae.

J Sci Food Agric. 2022-7

[10]
Multiplex PCR-Lateral Flow Dipstick Method for Detection of Thermostable Direct Hemolysin () Producing .

Biosensors (Basel). 2023-6-30

引用本文的文献

[1]
Development of Multiple Real-Time Fluorescent Quantitative PCR for Pathogen Detection in Aquaculture.

Vet Sci. 2025-4-2

[2]
Establishment and Application of Duplex Recombinase-Aided Amplification Combined with Lateral Flow Dipsticks for Rapid and Simultaneous Visual Detection of and in Milk.

Foods. 2025-2-9

[3]
Recombinase-aid amplification combined with lateral flow detection assay for sex identification of the great white pelican (Pelecanus onocrotalus).

Sci Rep. 2024-9-12

[4]
Rapid detection methods for foodborne pathogens based on nucleic acid amplification: Recent advances, remaining challenges, and possible opportunities.

Food Chem (Oxf). 2023-9-18

[5]
Recent Uses of Paper Microfluidics in Isothermal Nucleic Acid Amplification Tests.

Biosensors (Basel). 2023-9-15

本文引用的文献

[1]
A Fluorescent Recombinase Aided Amplification Assay for Detection of Babesia microti.

Korean J Parasitol. 2022-6

[2]
Development of an isothermal recombinase-aided amplification assay for the rapid and visualized detection of Klebsiella pneumoniae.

J Sci Food Agric. 2022-7

[3]
Automatic and multi-channel detection of bacteria on a slidable centrifugal disc based on FTA card nucleic acid extraction and recombinase aided amplification.

Lab Chip. 2021-12-21

[4]
Instrumentation-Compact Digital Microfluidic Reaction Interface-Extended Loop-Mediated Isothermal Amplification for Sample-to-Answer Testing of .

Anal Chem. 2021-7-20

[5]
Evaluation of loop-mediated isothermal amplification assay along with conventional and real-time PCR assay for sensitive detection of pathogenic Vibrio parahaemolyticus from seafood sample without enrichment.

Mol Biol Rep. 2021-1

[6]
RPA-Cas12a-FS: A frontline nucleic acid rapid detection system for food safety based on CRISPR-Cas12a combined with recombinase polymerase amplification.

Food Chem. 2020-7-19

[7]
Establishment of reverse transcription recombinase-aided amplification-lateral-flow dipstick and real-time fluorescence-based reverse transcription recombinase-aided amplification methods for detection of the Newcastle disease virus in chickens.

Poult Sci. 2020-7

[8]
Reverse-transcription recombinase-aided amplification assay for H7 subtype avian influenza virus.

Transbound Emerg Dis. 2019-12-4

[9]
Development and evaluation of a rapid and sensitive RPA assay for specific detection of Vibrio parahaemolyticus in seafood.

BMC Microbiol. 2019-8-13

[10]
A rapid and sensitive recombinase aided amplification assay incorporating competitive internal control to detect Bordetella pertussis using the DNA obtained by boiling.

Int J Infect Dis. 2019-7-6

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