A HCR based multivalent aptamer amplifier for ultrasensitive detection of Salmonella

Sun, Mengni; Ma, Na; Shi, Hanxing; Cheong, Ling‐Zhi; Yang, Wei; Qiao, Zhongliang

doi:10.1016/j.snb.2022.132860

Cited by 26 publications

(14 citation statements)

References 30 publications

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“…The binding affinities of TDN-multiApts with various valences were evaluated via equilibrium dissociation constants (Kds, inversely proportional to binding avidity) and measured by ELISA. 34 Briefly, 50 μL of target bacteria solution (10 5 cfu mL −1 ) was added into the high binding plates for a 2 h incubation. Afterward, 1% BSA was prepared and added into the plate at 37 °C for 30 min to block the redundant binding site.…”

Section: ■ Materials and Methodsmentioning

confidence: 99%

Dual-Functional Tetrahedron Multivalent Aptamer Assisted Amplification-Free CRISPR/Cas12a Assay for Sensitive Detection of Salmonella

Qiao,

Xue,

Sun

et al. 2023

J. Agric. Food Chem.

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Salmonellosis continues to impose a significant economic burden globally. Rapid and sensitive detection of Salmonella is crucial to preventing the outbreaks of foodborne illnesses, yet it remains a formidable challenge. Herein, a dual-functional tetrahedron multivalent aptamer assisted amplification-free CRISPR/Cas12a assay was developed for Salmonella detection. In the system, the aptamer was programmatically assembled on the tetrahedral DNA nanostructure to fabricate a multivalent aptamer (TDN-multiApt), which displayed a 3.5-fold enhanced avidity over the monovalent aptamer and possessed four CRISPR/Cas12a targeting fragments to amplify signal. Therefore, TDN-multiApt could directly activate Cas12a to achieve the second signal amplification without any nucleic acid amplification. By virtue of the synergism of high avidity and cascaded signal amplifications, the proposed method allowed the ultrasensitive detection of Salmonella as low as 7 cfu mL −1 . Meanwhile, this novel platform also exhibited excellent specificity against target bacteria and performed well in the detection of various samples, indicating its potential application in real samples.

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Section: ■ Materials and Methodsmentioning

confidence: 99%

Dual-Functional Tetrahedron Multivalent Aptamer Assisted Amplification-Free CRISPR/Cas12a Assay for Sensitive Detection of Salmonella

Qiao,

Xue,

Sun

et al. 2023

J. Agric. Food Chem.

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“…Notably, due to the low abundance, short size, high sequence homology among miRNAs family members, and susceptibility to degradation of miRNAs, detection of miRNAs with high sensitivity and specificity is challenging . Aiming to improve the sensitivity of miRNAs detection, various signal amplification systems have been developed, including nanomaterial-mediated amplification procedures [e.g., gold nanoparticles (GNPs), graphene nanosheets, and so on] − and molecular biological technique-based amplification strategies (e.g., hybridization chain reaction, strand-displacement amplification, and other DNA nanotechnology-based methods). − Among them, GNPs with excellent biocompatibility, facile preparation, and easy modification have been widely used in constructing signal amplification systems. ,,− Thermophoresis as an effective signal amplification approach utilizes the responses of particles to temperature gradient, thus inducing the motion and aggregation of particles, and finally achieving the signal amplification . Taking advantage of label-free and the capability of selectively driving suspended objects into warm or cold regions along a temperature gradient, thermophoresis has been employed for miRNA detection. , For example, Sun’s group reports a thermophoretic sensor for in situ detection of exosomal miRNAs with high accuracy, which relies on nanoflare detection of miRNAs and thermophoretic enrichment of cancer cell-derived exosomes .…”

Section: Introductionmentioning

confidence: 99%

Intracellular MicroRNA Imaging and Specific Discrimination of Prostate Cancer Circulating Tumor Cells Using Multifunctional Gold Nanoprobe-Based Thermophoretic Assay

Luo,

Meng,

et al. 2024

Anal. Chem.

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Circulating tumor cells (CTCs) have emerged as powerful biomarkers for diagnosis of prostate cancer. However, the effective identification and concurrently accurate imaging of CTCs for early screening of prostate cancer have been rarely explored. Herein, we reported a multifunctional gold nanoprobe-based thermophoretic assay for simultaneous specific distinguishing of prostate cancer CTCs and sensitive imaging of intracellular microRNA (miR-21), achieving the rapid and precise detection of prostate cancer. The multifunctional gold nanoprobe (GNP-DNA/Ab) was modified by two types of prostatespecific antibodies, anti-PSMA and anti-EpCAM, which could effectively recognize the targeting CTCs, and meanwhile linked double-stranded DNA for further visually imaging intracellular miR-21. Upon the specific internalization of GNP-DNA/Ab by PC-3 cells, target aberrant miR-21 could displace the signal strand to recover the fluorescence signal for sensitive detection at the single-cell level, achieving single PC-3 cell imaging benefiting from the thermophoresis-mediated signal amplification procedure. Taking advantage of the sensitive miR-21 imaging performance, GNP-DNA/Ab could be employed to discriminate the PC-3 and Jurkat cells because of the different expression levels of miR-21. Notably, PC-3 cells were efficiently recognized from white blood cells, exhibiting promising potential for the early diagnosis of prostate cancer. Furthermore, GNP-DNA/Ab possessed good biocompatibility and stability. Therefore, this work provides a great tool for aberrant miRNA-related detection and specific discrimination of CTCs, achieving the early and accurate diagnosis of prostate cancer.

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“…Complex pretreatment steps and a prolonged detection period contribute to these limitations. , Immunoassay techniques, exemplified by enzyme-linked immunosorbent assay (ELISA), offer specific and quick results but require enhanced sensitivity . Molecular biology assays provide speed and sensitivity but are hindered by the need for skilled operators, laboratory equipment, and the risk of false positives. , Biosensors, including antibodies and aptamers, − present advantages such as convenience, rapidity, high sensitivity, and specificity but also have drawbacks such as extended antibody preparation time, elevated costs, and limited stability. Aptamers face challenges with extended screening periods and susceptibilities to false positives.…”

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.

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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.

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A HCR based multivalent aptamer amplifier for ultrasensitive detection of Salmonella

Cited by 26 publications

References 30 publications

Dual-Functional Tetrahedron Multivalent Aptamer Assisted Amplification-Free CRISPR/Cas12a Assay for Sensitive Detection of Salmonella

Dual-Functional Tetrahedron Multivalent Aptamer Assisted Amplification-Free CRISPR/Cas12a Assay for Sensitive Detection of Salmonella

Intracellular MicroRNA Imaging and Specific Discrimination of Prostate Cancer Circulating Tumor Cells Using Multifunctional Gold Nanoprobe-Based Thermophoretic Assay

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

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