Immunocapture Magnetic Beads Enhanced the LAMP-CRISPR/Cas12a Method for the Sensitive, Specific, and Visual Detection of Campylobacter jejuni

Li, Chao; Chen, Xuan; Wen, Renqiao; Ma, Peng; Gu, Ke; Li, Cui; Zhou, Changyu; Lei, Changwei; Tang, Yizhi; Wang, Hongning

doi:10.3390/bios12030154

Cited by 27 publications

(11 citation statements)

References 55 publications

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“…We believe that this distinguishing feature will be a strong advantage for this technique. Beyond the demonstration of analytical applications, in the future, we hope that this technique will lead to the ability to rapidly synthesize large quantities of ssDNA , and could also be applied to isothermal amplification techniques. − …”

Section: Discussionmentioning

confidence: 99%

Biosensing with Polymerase Chain Reaction-Stable DNA-Functionalized Magnetically Susceptible Carbon–Iron Microparticles

Koirala,

Dalbec,

May

et al. 2023

Anal. Chem.

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We demonstrate a rapid and sensitive method for DNA detection without the need for fluorescence. This is based on carbon-coated magnetic iron (Fe) microparticles with a covalent surface attachment of DNA. We show that these magnetic microparticles can capture complementary DNA. Significantly, the DNA covalent surface bonds are robust to high temperatures and can be included in a sample during polymerase chain reaction (PCR). This method is employed for the detection of targeted DNA sequences (40–50 bp). Hybridization probes on the surface of the magnetically susceptible Fe microparticle recognize the target DNA sequence-specifically. The double-stranded DNA (dsDNA) microparticles are then quickly captured with a magnet from the sample matrix. This foregoes postpurification processes, such as electrophoresis, which make our technique time- and cost-effective. Captured dsDNA can be detected with intercalating dyes such as ethidium bromide through a loss in the UV absorption signal with a limit of detection (LOD) of 24 nM within 15 min. Likewise, surface-bound DNA can act as a primer in PCR to decrease the LOD to 5 pM within 2 h. This is the first instance of a nucleotide-modified magnetically susceptible carbon substrate that is PCR-compatible. Besides DNA capture, this strategy can eventually be applied to sequence-specific nucleic acid purification and enrichment, PCR cleanup, and single-strand generation. The DNA-coated particles are stable under PCR conditions (unlike commonly used polystyrene or gold particles).

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Section: Discussionmentioning

confidence: 99%

Biosensing with Polymerase Chain Reaction-Stable DNA-Functionalized Magnetically Susceptible Carbon–Iron Microparticles

Koirala,

Dalbec,

May

et al. 2023

Anal. Chem.

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“…Serological methods (i.e., immunoassays) (e.g., enzyme-linked immunosorbent assay; ELISA) [124][125][126][127][128], biosensors for the on-site detection of foodborne pathogens [129,130], DNA hybridization techniques (e.g., loop-mediated isothermal amplification; LAMP) [126,131,132], DNA fingerprinting techniques (e.g., multilocus sequence typing; MLST) [133][134][135] and above all PCR-based method and techniques (e.g., multiplex PCR; mPCR, quantitative or real time PCR; qPCR/rt-PCR), have been developed for the fastest and most efficient identification and differentiation of Campylobacter species among other foodborne pathogens. It should be noted though that some DNA fingerprinting techniques are more sophisticated (e.g., pulsed-field gel electrophoresis; PFGE, whole-genome sequencing; WGS) and require well-trained personnel with a know-how-to conduct the technique and interpret the data.…”

Section: Molecular Methods For Differentiating Campylobacter Speciesmentioning

confidence: 99%

Prevalence and Distribution of Thermotolerant Campylobacter Species in Poultry: A Comprehensive Review with a Focus on the Factors Affecting the Detection and Enumeration of Campylobacter jejuni and Campylobacter coli in Chicken Meat

Andritsos¹,

Tzimotoudis²,

Mataragas³

2023

Preprint

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It is well known that the strong-evidence foodborne outbreaks of human campylobacteriosis are associated with the consumption of raw or incompletely thermally processed poultry meat, whereas broilers act as the main reservoir for Campylobacter species. Campylobacter jejuni and Campylobacter coli are the two main species of campylobacters detected in chicken meat, while they account for almost 90% of the reported cases of campylobacteriosis in humans. Over 80% of these cases are attributed to C. jejuni and about 10% of them are due to C. coli. Therefore, until recently the dominance of C. jejuni against all other Campylobacter spp. isolated from chicken meat samples was well established and unquestionable. Lately, however, C. coli has been increasingly recovered from chicken meat to such an extent that it is now evident that it often comprises the dominant species among the identified campylobacters in the meat samples. This work attempts for the first time a detailed review in the literature to deepen into this noteworthy epidemiological swift in the prevalence of C. jejuni and C. coli, along with the distribution of Campylobacter spp. in chicken meat. Factors such as the sampling method followed for screening campylobacters in broiler carcasses (e.g., swabs or carcass rinsates, skinned or skinless meat excised samples) and part of the animal carcass from which the sample is obtained (e.g., neck, breast, leg), seasonality of sampling (summer vs. winter) and environmental conditions (e.g., rainfall, relative humidity) at the farm level, the isolation procedure (enumeration or detection) and pathogen identification (biochemical or molecular), the enrichment and plating isolation media (e.g., Bolton vs. Preston broth, charcoal-based vs. chromogenic agars), as well as the biofilm-forming ability of different campylobacters, highlight the multivariate dimension of the phenomenon and are thoroughly discussed in the present review.

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“…Additionally, a range of platforms have been developed by leveraging the advantages of CRISPR‐Cas with nucleic acid amplification technologies such as PCR (Ma, Peng, et al., 2021), RAA (Li, Ye, Chen, Xiang, et al., 2021), RPA (Chen et al., 2020; Zhuang et al., 2022), LAMP (Li, Chen, et al., 2022), multiple cross‐displacement amplification (Zhu et al., 2021), strand displacement amplification (Wang, Liu, et al., 2019; Zhou et al., 2018), rolling circle amplification (Qing et al., 2021), hybridization chain reaction (Liu et al., 2022), exponential amplification method (Huang et al., 2018; Tian et al., 2020), nicking enzyme‐assisted amplification (Bai et al., 2022), and so on. The CRISPR‐Cas biosensors are usually based on fluorescence signal readouts; other signal readouts such as colorimetry (Li, Zheng, et al., 2021), electrochemistry (Qing et al., 2021), lateral flow assay (Marsic et al., 2021), photothermal effect (Ma, Peng, et al., 2021), portable personal glucose meter (Liu, Hu, et al., 2021), surface‐enhanced Raman scattering (SERS) assay (Kim, Lee, Seo, et al., 2020; Pan et al., 2022), gas bubble signal (Silva et al., 2021), microfluidic paper‐based analytical device (μPAD) (Zhuang et al., 2022), hydrogel‐integrated paper‐based analytical device (μReaCH‐PAD) (Huang, Ni, et al., 2021), coupling biolayer interferometry (Qiao, Liu, et al., 2021), luminescence resonance energy transfer (Lin et al., 2022), and so on are established for visual and rapid biosensing applications.…”

Section: Crispr‐cas‐based Detectionmentioning

confidence: 99%

“…Most importantly, this method demonstrated a higher sensitivity (10 0 CFU/mL) and a shorter time (50 min) than conventional qPCR. Similarly, an immunocapture magnetic bead was used to capture the bacterial genomic DNA from complex samples, which was then released by heating (Li, Chen, et al., 2022). Organic‐aqueous solution was also used for extracting ochratoxin A, which was then detected using CRISPR‐Cas‐based assay (Mao, Wang, et al., 2022).…”

Section: Challenges and Opportunitiesmentioning

confidence: 99%

CRISPR‐Cas‐based detection for food safety problems: Current status, challenges, and opportunities

Man

Liu

2022

Comp Rev Food Sci Food Safe

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Food safety is one of the biggest public issues occurring around the world. Microbiological, chemical, and physical hazards can lead to food safety issues, which may occur at all stages of the supply chain. In order to tackle food safety issues and safeguard consumer health, rapid, accurate, specific, and field-deployable detection methods meeting diverse requirements are one of the imperative measures for food safety assurance. CRISPR-Cas system, a newly emerging technology, has been successfully repurposed in biosensing and has demonstrated huge potential to establish conceptually novel detection methods with high sensitivity and specificity. This review focuses on CRISPR-Cas-based detection and its current status and huge potential specifically for food safety inspection. We firstly illustrate the pending problems in food safety and summarize the popular detection methods. We then describe the potential applications of CRISPR-Casbased detection in food safety inspection. Finally, the challenges and futuristic

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Immunocapture Magnetic Beads Enhanced the LAMP-CRISPR/Cas12a Method for the Sensitive, Specific, and Visual Detection of Campylobacter jejuni

Cited by 27 publications

References 55 publications

Biosensing with Polymerase Chain Reaction-Stable DNA-Functionalized Magnetically Susceptible Carbon–Iron Microparticles

Biosensing with Polymerase Chain Reaction-Stable DNA-Functionalized Magnetically Susceptible Carbon–Iron Microparticles

Prevalence and Distribution of Thermotolerant <i>Campylobacter</i> Species in Poultry: A Comprehensive Review with a Focus on the Factors Affecting the Detection and Enumeration of <i>Campylobacter jejuni</i> and <i>Campylobacter coli</i> in Chicken Meat

CRISPR‐Cas‐based detection for food safety problems: Current status, challenges, and opportunities

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