Development of an RPA-based CRISPR/Cas12a assay in combination with a lateral flow strip for rapid detection of toxigenic Fusarium verticillioides in maize

Liang, Xiaoyan; Zhang, Xiu; Xi, Kaifei; Liu, Yang; Jijakli, M. Haissam; Guo, Wei

doi:10.1016/j.foodcont.2023.110172

Cited by 6 publications

(2 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Recombinase-assisted amplification (RAA), a novel nucleic acid thermostatic amplification technology, shares similarities with recombinase polymerase amplification (RPA). RAA achieves target signal amplification within 30 min under thermostatic conditions of 37–42 °C, making it particularly suitable for rapid detection and screening in the field …”

Section: Introductionmentioning

confidence: 99%

An Innovative and Efficient Fluorescent Detection Technique for Salmonella in Animal-Derived Foods Using the CRISPR/Cas12a-HCR System Combined with PCR/RAA

Wang,

Du,

Shao

et al. 2024

J. Agric. Food Chem.

View full text Add to dashboard Cite

Here, we present a method for Salmonella detection using clustered regularly interspaced short palindromic repeats associated with the CRISPR-associated protein 12a-hybridization chain reaction (CRISPR/Cas12a-HCR) system combined with polymerase chain reaction/recombinase-assisted amplification (PCR/RAA) technology. The approach relies on the Salmonella invA gene as a biorecognition element and its amplification through PCR and RAA. In the presence of the target gene, Cas12a, guided by crRNA, recognizes and cleaves the amplification product, initiating the HCR. Fluorescently labeled single-stranded DNA (ssDNA) H1 and H2 were introduced, and the Salmonella concentration was determined based on the fluorescence intensity from the triggered HCR. Both assays demonstrate high specificity, sensitivity, simplicity, and rapidity. The detection range was 2 × 10 1 −2 × 10 9 CFU/mL, with an LOD of 20 CFU/mL, and the entire process enabled specific and rapid Salmonella detection within 85−105 min. Field-incurred spiked recovery tests were conducted in mutton and beef samples using both assays, demonstrating satisfactory recovery and accuracy in animal-derived foods. By combining CRISPR/Cas12a with hybridization chain reaction technology, this study presents a rapid and sensitive Salmonella detection method that is crucial for identifying pathogenic bacteria and monitoring food safety.

show abstract

Section: Introductionmentioning

confidence: 99%

An Innovative and Efficient Fluorescent Detection Technique for Salmonella in Animal-Derived Foods Using the CRISPR/Cas12a-HCR System Combined with PCR/RAA

Wang,

Du,

Shao

et al. 2024

J. Agric. Food Chem.

View full text Add to dashboard Cite

show abstract

“…verticillioides) (teleomorph, Gibberella moniliformis) is a notorious maize pathogen that causes various disease in maize, including seedling blight, stalk rot, ear rot, and kernel rot in the global maize growing regions . Apart from causing yield losses, this fungus also produces a group of mycotoxins called fumonisins, which can contaminate maize kernels and pose a threat to food safety . More than a dozen types of fumonisins have been identified, including fumonisin A, fumonisin B, fumonisin C, and fumonisin P .…”

Section: Introductionmentioning

confidence: 99%

Development of an Efficient CRISPR/Cas9 System in Fusarium verticillioides and Its Application in Reducing Mycotoxin Contamination

Tang,

Ding,

Guo

2024

J. Agric. Food Chem.

Self Cite

View full text Add to dashboard Cite

Fusarium verticillioides (F. verticillioides) is a globally recognized and highly impactful fungal pathogen of maize, causing yield losses and producing harmful mycotoxins that pose a threat to human and animal health. However, the genetic tools available for studying this crucial fungus are currently limited in comparison to other important fungal pathogens. To address this, an efficient CRISPR/Cas9 genome editing system based on an autonomously replicating plasmid with an AMA1 sequence was established in this study. First, gene disruption of pyrG and pyrE via nonhomologous end-joining (NHEJ) pathway was successfully achieved, with efficiency ranging from 66 to 100%. Second, precise gene deletions were achieved with remarkable efficiency using a dual sgRNA expression strategy. Third, the developed genome editing system can be applied to generate designer chromosomes in F. verticillioides, as evidenced by the deletion of a crucial 38 kb fragment required for fumonisin biosynthesis. Fourth, the pyrG recycling system has been established and successfully applied in F. verticillioides. Lastly, the developed ΔFUM1 and ΔFUM mutants can serve as biocontrol agents to reduce the fumonisin B1 (FB1) contamination produced by the toxigenic strain. Taken together, these significant advancements in genetic manipulation and biocontrol strategies provide valuable tools for studying and mitigating the impact of F. verticillioides on maize crops.

show abstract

RT-RPA and RPA-LFA assay for rapid and ultrasensitive detection of Vibrio parahaemolyticus

Hu,

Chen,

Shi

et al. 2024

Food Control

View full text Add to dashboard Cite

Development of an RPA-based CRISPR/Cas12a assay in combination with a lateral flow strip for rapid detection of toxigenic Fusarium verticillioides in maize

Cited by 6 publications

References 33 publications

An Innovative and Efficient Fluorescent Detection Technique for Salmonella in Animal-Derived Foods Using the CRISPR/Cas12a-HCR System Combined with PCR/RAA

An Innovative and Efficient Fluorescent Detection Technique for Salmonella in Animal-Derived Foods Using the CRISPR/Cas12a-HCR System Combined with PCR/RAA

Development of an Efficient CRISPR/Cas9 System in Fusarium verticillioides and Its Application in Reducing Mycotoxin Contamination

RT-RPA and RPA-LFA assay for rapid and ultrasensitive detection of Vibrio parahaemolyticus

Contact Info

Product

Resources

About