Competitive fluorescence assay for Cd2+ based on aptamer structure-switching

Yu, Hao; Zhao, Qiang

doi:10.1016/j.microc.2023.109348

Cited by 3 publications

(3 citation statements)

References 32 publications

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“…As toxic metal pollutants, Cd 2+ can cause serious environmental damages and potential health risks. − Detection of Cd 2+ is of great significance to environmental monitoring and human health. , We further demonstrated the application of the CRISPR/Cas12a-amplified aptamer switch assay for Cd 2+ with the same principle by using an aptamer against Cd 2+ (CD17-T9-acDNA) − and the cDNA of the Cd 2+ aptamer (Table S1). We applied C13 for microplate coating because it is optimal for causing large signal changes in the presence of Cd 2+ . We optimized some important experimental conditions that affect the sensing performance, including NaCl concentration, MgCl 2 concentration, and incubation time.…”

Section: Resultsmentioning

confidence: 99%

“…We optimized some important experimental conditions that affect the sensing performance, including NaCl concentration, MgCl 2 concentration, and incubation time. Concentrations of NaCl and MgCl 2 in the binding buffer affected the binding affinity of the aptamer to Cd 2+ , and a low concentration of MgCl 2 promoted hybridization between the aptamer and cDNA. , Higher concentrations of NaCl and MgCl 2 were not suitable for sensitive Cd 2+ detection, resulting in a lower percentage of signal change (Figures S5 and S6), as they reduce the affinity of the aptamer for Cd 2+ . The incubation time of the mixture solution of the aptamer and Cd 2+ with cDNA immobilized on the microplate also affected the sensitivity of the assay.…”

Section: Resultsmentioning

confidence: 99%

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CRISPR/Cas12a-Amplified Aptamer Switch Microplate Assay for Small Molecules

Zhu,

Yu,

Zhao

2024

Anal. Chem.

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The presence of small molecule contaminants such as mycotoxins and heavy metals in foods and the environment causes a serious threat to human health and huge economic losses. The development of simple, rapid, sensitive, and on-site methods for small molecule pollutant detection is highly demanded. Here, combining the advantages of structure-switchable aptamer-mediated signal conversion and CRISPR/Cas12a-based signal amplification, we developed a CRISPR/Cas12a-amplified aptamer switch assay on a microplate for sensitive small molecule detection. In this assay, a short DNA strand complementary to the aptamer (cDNA) is immobilized on a microplate, which can capture the aptamer-linked active DNA probe (Apt-acDNA) in the sample solution when the target is absent. With the addition of the Cas12a reporter system, the captured Apt-acDNA probes activate Cas12a to indiscriminately cleave fluorescent DNA substrates, producing a high fluorescence signal. When the target is present, the Apt-acDNA probe specifically binds to the target rather than hybridizing with cDNA on the microplate, and the fluorescence signal is reduced. The analytical performance of our method was demonstrated by the detection of two highly toxic pollutants, aflatoxin B1 (AFB1) and cadmium ion (Cd2+), as examples. The assay exhibited good selectivity and high sensitivity, with detection limits of 31 pM AFB1 and 3.9 nM Cd2+. It also allowed the detection of targets in the actual sample matrix. With the general signal conversion strategy, this method can be used to detect other targets by simply changing the aptamer and cDNA, showing potential practical applications in broad fields.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

CRISPR/Cas12a-Amplified Aptamer Switch Microplate Assay for Small Molecules

Zhu,

Yu,

Zhao

2024

Anal. Chem.

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“…A flexible and cost-effective design allows aptamers to be used in analysis methods with excellent performance [10]. Detection methods using aptamers have been employed in conjunction with colorimetry [11], fluorescence [12,13], surface plasmon resonance [14,15], and chemiluminescent [16] and electrochemical [17][18][19] techniques. The literature shows a growing number of studies aimed at the development of electrochemical biosensors based on aptamers (Figure 2).…”

Section: Introductionmentioning

confidence: 99%

Relevant Aspects in the Development of Electrochemical Aptasensors for the Determination of Antibiotics—A Review

da Silva,

Pereira

2023

Electrochem

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Aptamers are three-dimensional structures of DNA or RNA that present high affinity and selectivity to specific targets, obtained through in vitro screening. Aptamers are used as biological recognizers in electrochemical biosensors, the so-called aptasensors, providing greater specificity in recognizing the most diverse analytes. Electrochemical aptasensors have extremely relevant characteristics, such as high sensitivity, low cost compared to other biorecognizers such as antibodies, and excellent compatibility, being considered one of the most promising alternative methods in several areas, such as biomedical diagnosis and monitoring environmental contaminants. In this sense, the present work reviews the relevant aspects of methodologies based on electrochemical aptasensors and their applications in determining antibiotics, seeking to foster innovation in electrochemical biosensors.

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Aptasensors and Advancement in Molecular Recognition Technology

Napit,

Jaysawal,

Chowdhury

et al. 2024

Adv Materials Technologies

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Synthetic oligonucleic acids known as aptamers exhibit remarkable selectivity and affinity for target recognition and binding. Selected via an iterative process known as “selective evolution of ligands by exponential enrichment” (SELEX), aptamers fold into defined 3D conformations to interact with their targets. The incorporation of aptamers as recognition elements has driven notable progress in biosensors, giving rise to the development of aptasensors. Here, the process of aptamer discovery and the development of various types of aptasensors are summarized. The fundamental design principles of aptasensors are elaborated along with the superiority of aptamers compared to antibodies. The various modes employed by aptasensors, such as structure‐switching design, hybridization chain reaction amplification, enzyme‐assisted recycling, and split aptamer design are examined. Further light is shed on the diverse landscape of aptasensors, their adaptability to different analytes aptasensors as well as their potential to propel advancements in modern biosensor technology. As a nucleic acids‐based biosensor platform, aptasensors poise to become a next generation of sensitive and cost‐effective technology to shape the future of molecular recognition in biosensing.

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Competitive fluorescence assay for Cd2+ based on aptamer structure-switching

Cited by 3 publications

References 32 publications

CRISPR/Cas12a-Amplified Aptamer Switch Microplate Assay for Small Molecules

CRISPR/Cas12a-Amplified Aptamer Switch Microplate Assay for Small Molecules

Relevant Aspects in the Development of Electrochemical Aptasensors for the Determination of Antibiotics—A Review

Aptasensors and Advancement in Molecular Recognition Technology

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