Harnessing enhanced CRISPR/Cas12a trans-cleavage activity with extended reporters and reductants for early diagnosis of Helicobacter pylori, the causative agent of peptic ulcers and stomach cancer

Habimana, Jean de Dieu; Mukama, Omar; Chen, Guiquan; Chen, Mengjun; Amissah, Obed Boadi; Wang, Lin; Liu, Yujie; Sun, Yuhan; Li, Amy L.; Deng, Sihao; Huang, Jufang; Yan, Xiao‐Xin; Rutaganda, Theobard; Mutangana, Dieudonné; Wu, Lin; Huang, Rongqi; Li, Zhiyuan

doi:10.1016/j.bios.2022.114939

Cited by 17 publications

(4 citation statements)

References 48 publications

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“…The patients were examined at the end of treatment and the treatment effect was evaluated in terms of inflammation, symptoms and signs, which were divided into three parts like significant effect-The active inflammation of gastric mucosa disappeared in patients and clinical symptoms and signs disappeared, and the results of 13 C urea breath test were negative in patients after treatment [8] ; effective-Microscopically visible gastric mucosal lesions were reduced by half compared with those before treatment and clinical symptoms and the 13 C urea breath test result was negative or changed from strong positive to weak positive; invalid-The clinical symptoms and signs did not improve after treatment, and the 13 C urea breath test result was still positive after treatment.…”

Section: Observation Indexes and Evaluation Methodsmentioning

confidence: 99%

Effect of Triple Therapy with Lactobacillus acidophilus Tablets on Intestinal Flora in Chronic Helicobacter pylori Infected Atrophic Gastritis

Dai,

Cao,

Weng

et al. 2024

IJPS

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Dai et al.: Effect of Lactobacillus acidophilus Tablets on Helicobacter pylori Infected Atrophic GastritisTo evaluate the effects of Lactobacillus acidophilus tablets based on quadruple therapy on intestinal flora and inflammatory response in patients with Helicobacter pylori-infected chronic atrophic gastritis is the objective of the study. The clinical data of Helicobacter pylori-infected patients with Helicobacter pylori-infected chronic atrophic gastritis were analyzed retrospectively and 90 patients who met the criteria of inclusion and exclusion were selected as subjects. The control group was treated with conventional quadruple therapy and the observation group was treated with compound Lactobacillus acidophilus tablets combined with quadruple therapy. The intestinal flora and inflammation of the patients were compared. The efficacy of 82 % (41/50) and 96 % (p<0.05) were significantly higher in the post-treatment observation group than in the control group; the area of gastric lesion (0.85±0.17) was significantly lower than in the control group (p<0.05) and the digestive system symptoms score (3.23±1.13) was significantly lower than in the control group (6.67 %). Gastrointestinal function score (4.63±1.39) and Helicobacter pylori eradication rate (47/50) were significantly higher (94 %) in the observation group than in the control group, p<0.05; tumor necrosis factor alpha, (23.39±6.20 µg/ml) and interleukin-6 (15.39±6.41 µg/ml) were significantly lower than that in the control group and gastrin level (1.78±0.19 nmol/l) was significantly higher. Compound Lactobacillus acidophilus tablets based on quadruple therapy for the treatment of Helicobacter pylori-infected chronic atrophic gastritis can well eradicate Helicobacter pylori, control intestinal inflammation, improve intestinal flora and reduce clinical symptoms and signs of patients.

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Section: Observation Indexes and Evaluation Methodsmentioning

confidence: 99%

Effect of Triple Therapy with Lactobacillus acidophilus Tablets on Intestinal Flora in Chronic Helicobacter pylori Infected Atrophic Gastritis

Dai,

Cao,

Weng

et al. 2024

IJPS

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“…The LbCas12a-mediated trans -cleavage assay was carried out following a method with slight modification . A 20 μL mixture consisted of 10 μL of 2× Buffer, 0.8 μM Cas12a (1 μL), 10 μM reporters (1 μL) (Table S2), 1.2 μM crRNA (1 μL), 100 mM MgSO 4 (1 μL), 2 μL of activator, and DEPC water to make up the final volume. A portable Dhelix-Q5 isothermal fluorescence PCR machine was used for real-time monitoring of trans -cleavage activity at 37 °C for 30–60 min.…”

Section: Methodsmentioning

confidence: 99%

A Rationally Designed CRISPR/Cas12a Assay Using a Multimodal Reporter for Various Readouts

et al. 2023

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The CRISPR/Cas systems offer a programmable platform for nucleic acid detection, and CRISPR/Cas-based diagnostics (CRISPR-Dx) have demonstrated the ability to target nucleic acids with greater accuracy and flexibility. However, due to the configuration of the reporter and the underlying labeling mechanism, almost all reported CRISPR-Dx rely on a single-option readout, resulting in limitations in end-point result readouts. This is also associated with high reagent consumption and delays in diagnostic reports due to protocol differences. Herein, we report for the first time a rationally designed Cas12a-based multimodal universal reporter (CAMURE) with improved sensitivity that harnesses a dual-mode reporting system, facilitating options in endpoint readouts. Through systematic configurations and optimizations, our novel universal reporter achieved a 10-fold sensitivity enhancement compared to the DETECTR reporter. Our unique and versatile reporter could be paired with various readouts, conveying the same diagnostic results. We applied our novel reporter for the detection of staphylococcal enterotoxin A due to its high implication in staphylococcal food poisoning. Integrated with loop-mediated isothermal amplification, our multimodal reporter achieved 10 CFU/mL sensitivity and excellent specificity using a real-time fluorimeter, in-tube fluorescence, and lateral flow strip readouts. We also propose, using artificially contaminated milk samples, a fast (2−5 min) Triton X-100 DNA extraction approach with a comparable yield to the commercial extraction kit. Our CAMURE could be leveraged to detect all gene-encoding SEs by simply reprogramming the guide RNA and could also be applied to the detection of other infections and disease biomarkers.

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“…Polyphenols such as kaempferol inhibit reproductive enzymes, destabilize the cytoplasmic membrane, and regulate MAPK and NF-κB pathways to mitigate inflammatory damage ( Bi et al, 2022 ). Alkaloids, such as coptisine, inhibit urease activity and disrupt cell membranes ( Habimana et al, 2023 ). Combining plant foods with antibiotics enhances H. pylori eradication and offers potential for preventing associated gastric disorders ( Liu et al, 2023 ).…”

Section: New Approachesmentioning

confidence: 99%

Novel therapeutic regimens against Helicobacter pylori: an updated systematic review

Huang,

Cao,

Cao

2024

Front. Microbiol.

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Helicobacter pylori (H. pylori) is a strict microaerophilic bacterial species that exists in the stomach, and H. pylori infection is one of the most common chronic bacterial infections affecting humans. Eradicating H. pylori is the preferred method for the long-term prevention of complications such as chronic gastritis, peptic ulcers, gastric mucosa-associated lymphoid tissue lymphoma, and gastric cancer. However, first-line treatment with triple therapy and quadruple therapy has been unable to cope with increasing antibacterial resistance. To provide an updated review of H. pylori infections and antibacterial resistance, as well as related treatment options, we searched PubMed for articles published until March 2024. The key search terms were “H. pylori”, “H. pylori infection”, “H. pylori diseases”, “H. pylori eradication”, and “H. pylori antibacterial resistance.” Despite the use of antimicrobial agents, the annual decline in the eradication rate of H. pylori continues. Emerging eradication therapies, such as the development of the new strong acid blocker vonoprazan, probiotic adjuvant therapy, and H. pylori vaccine therapy, are exciting. However, the effectiveness of these treatments needs to be further evaluated. It is worth mentioning that the idea of altering the oxygen environment in gastric juice for H. pylori to not be able to survive is a hot topic that should be considered in new eradication plans. Various strategies for eradicating H. pylori, including antibacterials, vaccines, probiotics, and biomaterials, are continuously evolving. A novel approach involving the alteration of the oxygen concentration within the growth environment of H. pylori has emerged as a promising eradication strategy.

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Harnessing enhanced CRISPR/Cas12a trans-cleavage activity with extended reporters and reductants for early diagnosis of Helicobacter pylori, the causative agent of peptic ulcers and stomach cancer

Cited by 17 publications

References 48 publications

Effect of Triple Therapy with Lactobacillus acidophilus Tablets on Intestinal Flora in Chronic Helicobacter pylori Infected Atrophic Gastritis

Effect of Triple Therapy with Lactobacillus acidophilus Tablets on Intestinal Flora in Chronic Helicobacter pylori Infected Atrophic Gastritis

A Rationally Designed CRISPR/Cas12a Assay Using a Multimodal Reporter for Various Readouts

Novel therapeutic regimens against Helicobacter pylori: an updated systematic review

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