scholarly journals Simple Visualized Detection Method of Virulence-Associated Genes of Vibrio cholerae by Loop-Mediated Isothermal Amplification

2019 ◽  
Vol 10 ◽  
Author(s):  
Mengjie Xu ◽  
Huiyu Fu ◽  
Dailing Chen ◽  
Zehuai Shao ◽  
Jun Zhu ◽  
...  
Keyword(s):  
Vibrio Cholerae ◽  
Detection Method ◽  
Isothermal Amplification ◽  
Loop Mediated Isothermal Amplification ◽  
Visualized Detection

scholarly journals Rapid and Visualized Detection of Virulence-Related Genes of Vibrio cholerae in Water and Aquatic Products by Loop-Mediated Isothermal Amplification

2021 ◽  
Author(s):  
DAILING CHEN ◽  
ZHILI LIANG ◽  
SHUNLIN REN ◽  
WALID ALALI ◽  
Lanming Chen
Keyword(s):  
Vibrio Cholerae ◽  
Target Genes ◽  
Limit Of Detection ◽  
Molecular Probes ◽  
Isothermal Amplification ◽  
Lamp Method ◽  
Loop Mediated Isothermal Amplification ◽  
Aquatic Products ◽  
Standard Polymerase Chain Reaction ◽  
Visualized Detection

Vibrio cholerae  can cause pandemic cholera in humans. The bacterium resides in aquatic environments worldwide. Continuous testing of V. cholerae contamination in water and aquatic products is imperative for food safety control and human health. In this study, a rapid and visualized method was for the first time developed based on loop-mediated isothermal amplification (LAMP) for detection of very important virulence-related genes ace , zot , cri , and  nanH for toxins and infection process of V. cholerae . Three pairs of molecular probes targeting each of these genes were designed and synthesized. The one-step LAMP reaction was conducted at 65 o C for 40 min. Positive results were simply inspected by the production of light green color under visible light or green fluorescence under UV light (302 nm). Limit of detection (LOD) of the LAMP method ranged from 1.85-2.06 pg/reaction of genomic DNA or 2.50-4.00×10 2  CFU/reaction for target genes of cell culture of V. cholerae , which was more sensitive than standard polymerase chain reaction (PCR). Inclusivity and exclusivity of the LAMP method were 100% for all target genes. The method showed similar high efficiency to a certain extent in rapid testing of spiked or collected specimens of water and aquatic products. Target genes were detected by the absence from all water samples from various sources. However, high occurrences of nanH  gene were observed in intestine samples derived from four species of fish and one species of shellfish, indicating a risk of potentially toxic V. cholerae  in commonly consumed aquatic products. The results in this study provide a potential tool for rapid and visualized detection of V. cholerae in water and aquatic products.

Development of a loop-mediated isothermal amplification assay for Vibrio cholerae O1 and O139

2019 ◽  
Vol 45 ◽  
pp. 65-67 ◽  
Author(s):  
Hidemasa Izumiya ◽  
Masatomo Morita ◽  
Eiji Arakawa ◽  
Tuan Cuong Ngo ◽  
Hoai Thu Nguyen ◽  
...  
Keyword(s):  
Vibrio Cholerae ◽  
Isothermal Amplification ◽  
Loop Mediated Isothermal Amplification ◽  
Amplification Assay ◽  
Vibrio Cholerae O1

scholarly journals The Development of Highly Specific and Sensitive Primers for the Detection of Potentially Allergenic Soybean (Glycine max) Using Loop-Mediated Isothermal Amplification Combined with Lateral Flow Dipstick (LAMP-LFD)

Foods ◽  
2020 ◽  
Vol 9 (4) ◽  
pp. 423 ◽  
Author(s):  
Stefanie M. Allgöwer ◽  
Chris A. Hartmann ◽  
Thomas Holzhauser
Keyword(s):  
Glycine Max ◽  
Detection Method ◽  
Lateral Flow ◽  
Isothermal Amplification ◽  
Loop Mediated Isothermal Amplification ◽  
Lateral Flow Dipstick ◽  
Soybean Allergy ◽  
Systematic Development ◽  
Multicopy Genes ◽  
Selection Of

The soybean (Glycine max) has been recognized as a frequent elicitor of food allergy worldwide. A lack of causative immunotherapy of soybean allergy makes soybean avoidance essential. Therefore, sensitive and specific methods for soybean detection are needed to allow for soybean verification in foods. Loop-mediated isothermal amplification (LAMP) represents a rapid and simple DNA-based detection method principally suitable for field-like applications or on-site analytical screening for allergens during the manufacturing of foods. This work describes the systematic development and selection of suitable LAMP primers based on soybean multicopy genes. The chemistry applied allows for a versatile detection of amplified DNA, using either gel electrophoresis, fluorescence recording, or a simple Lateral Flow Dipstick (LFD). LAMP based on the ORF160b gene was highly specific for the soybean and may allow for a detection level equivalent to approximately 10 mg soy per kg food. Various soybean cultivars were detectable at a comparable level of sensitivity. LAMP combined with LFD-like detection facilitates a simple, highly specific and sensitive detection of the soybean without the need for expensive analytical equipment. In contrast to the majority of antibody-based methods for soybean detection, all identified primer sequences and optimized protocols are disclosed and broadly available to the community.

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