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

Xu, Mengjie; Fu, Huiyu; Chen, Dailing; Shao, Zhigang; Zhu, Jun; Alali, Walid Q.; Chen, Lanming

doi:10.3389/fmicb.2019.02899

Cited by 14 publications

(33 citation statements)

References 70 publications

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“…Bacterial strains and culture media used in this study are listed in Table S1. Bacterial culture media were purchased as described previously [28]. Vibrio strains were inoculated in media (pH 8.4-8.5) with 3.0% NaCl, while non-Vibrios in media (pH 7.0-7.2) with 1% NaCl [28].…”

Section: Bacterial Strains and Culture Conditionsmentioning

confidence: 99%

Identification of Antibacterial Components in the Methanol-Phase Extract from Edible Herbaceous Plant Rumex madaio Makino and Their Antibacterial Action Modes

Liu

Yang

et al. 2022

Molecules

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Outbreaks and prevalence of infectious diseases worldwide are some of the major contributors to morbidity and morbidity in humans. Pharmacophageous plants are the best source for searching antibacterial compounds with low toxicity to humans. In this study, we identified, for the first time, antibacterial components and action modes of methanol-phase extract from such one edible herbaceous plant Rumex madaio Makino. The bacteriostatic rate of the extract was 75% against 23 species of common pathogenic bacteria. The extract was further purified using the preparative high-performance liquid chromatography (Prep-HPLC) technique, and five separated componential complexes (CC) were obtained. Among these, the CC 1 significantly increased cell surface hydrophobicity and membrane permeability and decreased membrane fluidity, which damaged cell structure integrity of Gram-positive and -negative pathogens tested. A total of 58 different compounds in the extract were identified using ultra-HPLC and mass spectrometry (UHPLC-MS) techniques. Comparative transcriptomic analyses revealed a number of differentially expressed genes and various changed metabolic pathways mediated by the CC1 action, such as down-regulated carbohydrate transport and/or utilization and energy metabolism in four pathogenic strains tested. Overall, the results in this study demonstrated that the CC1 from R. madaio Makino are promising candidates for antibacterial medicine and human health care products.

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Section: Bacterial Strains and Culture Conditionsmentioning

confidence: 99%

Identification of Antibacterial Components in the Methanol-Phase Extract from Edible Herbaceous Plant Rumex madaio Makino and Their Antibacterial Action Modes

Liu

Yang

et al. 2022

Molecules

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“…Thus, LAMP has achieved wide recognition (Law et al 2015). Law et al (2016) compared a variety of rapid methods used for detecting foodborne bacterial pathogens, and concluded that LAMP is clearly an important method for rapid detection and control of spread of foodborne pathogens and prevention of foodborne disease outbreaks Xu et al (2019) developed their LAMP method and compared it with PCR assay, finding LAMP detected the virulence genes hapA, mshA, pilA and tlh, and also the toxigenic genes ctxA and tcpA, as well as the species-specific gene lolB of V.…”

Section: Discussionmentioning

confidence: 99%

Loop-mediated isothermal amplification (LAMP) assay for rapid detection of viable but non-culturable Vibrio cholerae O1

2022

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Vibrio cholerae, an important waterborne pathogen, is a rod-shaped bacterium that naturally exists in aquatic environments. When conditions are unfavorable for growth, the bacterium can undergo morphological and physiological changes to assume a coccoid morphology. This stage in its life cycle is referred to as viable but non-culturable (VBNC) since VBNC cells do not grow on conventional bacteriological culture media. The current study compared polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) to detect and identify VBNC V. cholerae. Because it is difficult to detect and identify VBNC V. cholerae, the results of the current study are useful in showing LAMP to be more sensitive and rapid than PCR in detecting and identifying non-culturable, coccoid forms of V. cholerae. Furthermore, the LAMP method is effective in detecting and identifying very low numbers of coccoid VBNC V. cholerae in environmental water samples, with the added benefit of being inexpensive to perform.

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“…V. cholerae isolates were individually incubated in 5 mL of tryptic soy broth (TSB) (Land Bridge, Beijing, China) (pH 8.5, 3.0% NaCl) at 37°C for 16–18 h. Genomic DNA of the isolates was prepared using the TaKaRa MiniBEST Bacterial Genomic DNA Extraction Kit Ver.3.0 (TaKaRa Biomedical Technology Co., Ltd., Beijing, China) following the manufacturer’s instructions. Extracted DNA samples were analyzed as described previously [ 47 ].…”

Section: Methodsmentioning

confidence: 99%

“…PCRs were performed as described previously [ 6 ]. Genomic DNA of V. cholerae ATCC 14035 ( ctxAB + , tcpA + , ace + , and zot + ) and GIM 1.449 ( ctxAB − , rtxCABD + , hlyA + , tlh + , and hapA + ) were used as positive controls as described previously [ 6 , 47 ]. Amplicons were confirmed by DNA sequencing at Sangon (Shanghai, China).…”

Section: Methodsmentioning

confidence: 99%

First Experimental Evidence for the Presence of Potentially Toxic Vibrio cholerae in Snails, and Virulence, Cross-Resistance and Genetic Diversity of the Bacterium in 36 Species of Aquatic Food Animals

Chen

et al. 2021

Antibiotics

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Vibrio cholerae is the most common waterborne pathogen that can cause pandemic cholera in humans. Continuous monitoring of V. cholerae contamination in aquatic products is crucial for assuring food safety. In this study, we determined the virulence, cross-resistance between antibiotics and heavy metals, and genetic diversity of V. cholerae isolates from 36 species of aquatic food animals, nearly two-thirds of which have not been previously detected. None of the V. cholerae isolates (n = 203) harbored the cholera toxin genes ctxAB (0.0%). However, isolates carrying virulence genes tcpA (0.98%), ace (0.5%), and zot (0.5%) were discovered, which originated from the snail Cipangopaludina chinensis. High occurrences were observed for virulence-associated genes, including hapA (73.4%), rtxCABD (68.0–41.9%), tlh (54.2%), and hlyA (37.9%). Resistance to moxfloxacin (74.9%) was most predominant resistance among the isolates, followed by ampicillin (59.1%) and rifampicin (32.5%). Approximately 58.6% of the isolates displayed multidrug resistant phenotypes. Meanwhile, high percentages of the isolates tolerated the heavy metals Hg2+ (67.0%), Pb2+ (57.6%), and Zn2+ (57.6%). Distinct virulence and cross-resistance profiles were discovered among the V. cholerae isolates in 13 species of aquatic food animals. The ERIC-PCR-based genome fingerprinting of the 203 V. cholerae isolates revealed 170 ERIC-genotypes, which demonstrated considerable genomic variation among the isolates. Overall, the results of this study provide useful data to fill gaps for policy and research related to the risk assessment of V. cholerae contamination in aquatic products.

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

Cited by 14 publications

References 70 publications

Identification of Antibacterial Components in the Methanol-Phase Extract from Edible Herbaceous Plant Rumex madaio Makino and Their Antibacterial Action Modes

Identification of Antibacterial Components in the Methanol-Phase Extract from Edible Herbaceous Plant Rumex madaio Makino and Their Antibacterial Action Modes

Loop-mediated isothermal amplification (LAMP) assay for rapid detection of viable but non-culturable Vibrio cholerae O1

First Experimental Evidence for the Presence of Potentially Toxic Vibrio cholerae in Snails, and Virulence, Cross-Resistance and Genetic Diversity of the Bacterium in 36 Species of Aquatic Food Animals

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