Using Recombinase-Aid Amplification Combined with Pyrococcus furiosus Argonaute for Rapid Sex Identification in Flamingo (Phoenicopteridae) DOI Creative Commons
Shuangshuang Tan, Fanwen Zeng,

Wanhuan Zhong

et al.

Animals, Journal Year: 2024, Volume and Issue: 15(1), P. 7 - 7

Published: Dec. 24, 2024

Flamingos (Phoenicopteridae) are among the oldest birds worldwide and loved by people for their bright red feathers. In addition, flamingos sexually monomorphic birds, distinguishing between males females is difficult. The polymerase chain reaction (PCR) widely used sex identification. However, PCR method requires a precise thermal cycler in laboratory time-consuming. Therefore, developing rapid, sensitive, accurate to identify of crucial. this study, we established identification system using recombinase-aided amplification-Pyrococcus furiosus Argonaute (RAA-PfAgo) technique greater flamingo (Phoenicopterus roseus). flamingo-RAA-PfAgo can unknown-sex less than 1 h be visualized fluorescent detector or blue light. results showed that optimal RAA-PfAgo conditions could detect 0.6 ng genomic DNA effectively differentiate females. Random sample evaluations revealed had 100% coincidence rate compared with conventional PCR. conclusion, study provides specific, reference sexing.

Language: Английский

CRISPR/Cas14 and G-Quadruplex DNAzyme-Driven Biosensor for Paper-Based Colorimetric Detection of African Swine Fever Virus DOI

Xue Zhao,

Yawen He,

Shengjie Shao

et al.

ACS Sensors, Journal Year: 2024, Volume and Issue: 9(5), P. 2413 - 2420

Published: April 18, 2024

The highly contagious nature and 100% fatality rate contribute to the ongoing expanding impact of African swine fever virus (ASFV), causing significant economic losses worldwide. Herein, we developed a cascaded colorimetric detection using combination CRISPR/Cas14a system, G-quadruplex DNAzyme, microfluidic paper-based analytical device. This CRISPR/Cas14a-G4 biosensor could detect ASFV as low 5 copies/μL differentiate wild-type mutated DNA with 2-nt difference. Moreover, this approach was employed in porcine plasma. A broad linear range observed, limit spiked plasma calculated be 42–85 copies/μL. Our results indicate that paper platform exhibits advantages high sensitivity, excellent specificity, cost, making it promising for clinical applications field disease suitable popularization low-resourced areas.

Language: Английский

Citations

19
Micropore Resistance Counting Platform for Multiplexed and Ultrasensitive Detection of Mycotoxins and Biomarkers DOI
Letian Li, Mengjiao Wang, Yongzhen Dong

et al.

ACS Nano, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 3, 2025

Development of a multiplexed and sensitive biosensing platform is priority for public health security. We report micropore resistance counting based on polystyrene microsphere size-based encoding Clostridium butyricum Argonaute (CbAgo) decoding ultrasensitive detection. Initially, we constructed target DNA-modified coding system counting. Subsequently, the precise recognition cleavage capabilities guide DNA-activated CbAgo protein enable encoded system. Changes in concentration microspheres are presented as signal readout. The demonstrated excellent performance detection three mycotoxins (with sensitivity range over 4 orders magnitude reaching pg/mL level) two inflammatory markers at pg/mL. Combining enzyme by with counting, developed highly tool wide-ranging potential applications such clinical diagnosis, food safety inspection, environmental monitoring.

Language: Английский

Citations

2
Machine Learning-Assistant Colorimetric Sensor Arrays for Intelligent and Rapid Diagnosis of Urinary Tract Infection DOI
Jianyu Yang, Ge Li, Shihong Chen

et al.

ACS Sensors, Journal Year: 2024, Volume and Issue: 9(4), P. 1945 - 1956

Published: March 26, 2024

Urinary tract infections (UTIs), which can lead to pyelonephritis, urosepsis, and even death, are among the most prevalent infectious diseases worldwide, with a notable increase in treatment costs due emergence of drug-resistant pathogens. Current diagnostic strategies for UTIs, such as urine culture flow cytometry, require time-consuming protocols expensive equipment. We present here machine learning-assisted colorimetric sensor array based on recognition ligand-functionalized Fe single-atom nanozymes (SANs) identification microorganisms at order, genus, species levels. Colorimetric arrays built from SAN Fe1–NC functionalized four types ligands, generating unique microbial fingerprints. By integrating trained computational classification model, platform identify more than 10 UTI samples within 1 h. Diagnostic accuracy up 97% was achieved 60 clinical samples, holding great potential translation into practice applications.

Language: Английский

Citations

14
Advances of engineered microfluidic biosensors via CRISPR/Cas in bacteria and virus monitoring DOI
Xintong Li,

Tingya Wang,

Xiaohui Liu

et al.

Chemical Engineering Journal, Journal Year: 2024, Volume and Issue: 491, P. 152038 - 152038

Published: May 8, 2024

Language: Английский

Citations

7
On-Site and Visual Detection of the H5 Subtype Avian Influenza Virus Based on RT-RPA and CRISPR/Cas12a DOI Creative Commons

Xu Zhou,

Siwen Wang, Yue Ma

et al.

Viruses, Journal Year: 2024, Volume and Issue: 16(5), P. 753 - 753

Published: May 10, 2024

Avian influenza viruses (AIVs) of the H5 subtype rank among most serious pathogens, leading to significant economic losses in global poultry industry and posing risks human health. Therefore, rapid accurate virus detection is crucial for prevention control AIVs. In this study, we established a novel method by utilizing precision CRISPR/Cas12a efficiency RT-RPA technologies. This assay facilitates direct visualization results through blue light lateral flow strips, accurately identifying with high specificity without cross-reactivity against other AIV subtypes, NDV, IBV, IBDV. With thresholds 1.9 copies/μL (blue light) × 103 (lateral strips), our not only competes but also slightly surpasses RT-qPCR, demonstrating an 80.70% positive rate across 81 clinical samples. The RT-RPA/CRISPR-based characterized sensitivity, specificity, independence from specialized equipment. immediate field applicability RT-RPA/CRISPR approach underscores its importance as effective tool early management outbreaks caused

Language: Английский

Citations

7
Nanophotonic catheters: A lens into the body for biosensing and biomedical imaging DOI
Bakr Ahmed Taha,

Inas A. Al-Tahar,

Ali J. Addie

et al.

Applied Materials Today, Journal Year: 2024, Volume and Issue: 38, P. 102229 - 102229

Published: May 21, 2024

Language: Английский

Citations

7
PfAgo-Based Zika Virus Detection DOI Creative Commons
Yuhao Chen,

Xianyi Zhang,

Xuan Yang

et al.

Viruses, Journal Year: 2024, Volume and Issue: 16(4), P. 539 - 539

Published: March 30, 2024

As a mosquito-borne flavivirus, Zika virus (ZIKV) has been identified as global health threat. The linked to severe congenital disabilities, including microcephaly and other malformations, resulting in fatal intrauterine death. Therefore, developing sensitive specific methods for the early detection accurate diagnosis of ZIKV is essential controlling its spread mitigating impact on public health. Herein, we set up novel nucleic acid system based Pyrococcus furiosus Argonaute (PfAgo)-mediated detection, targeting non-structural protein 5 (NS5) region genome (abbreviated ZIKV-PAND). Without preamplification with polymerase chain reaction (PCR), minimum concentration (MDC) ZIKV-PAND was about 10 nM. When introducing an amplification step, MDC can be dramatically decreased aM level (8.3 aM), which comparable qRT-PCR assay (1.6 aM). In addition, diagnostic findings from analysis simulated clinical samples or using show complete agreement 100% assays. This correlation aid implementation molecular testing diagnoses investigation infection epidemiological scale.

Language: Английский

Citations

5
Argonaute-Based Nucleic Acid Detection Technology: Advantages, Current Status, Challenges, and Perspectives DOI
Yaru Li, Lu Zhao,

Jiali Wang

et al.

ACS Sensors, Journal Year: 2024, Volume and Issue: 9(11), P. 5665 - 5682

Published: Nov. 11, 2024

Rapid and accurate detection is a prerequisite for precise clinical diagnostics, ensuring food safety, facilitating biotechnological applications. The Argonaute system, as cutting-edge technique, has been successfully repurposed in biosensing beyond the CRISPR/Cas system (clustered regularly interspaced short palindromic repeats CRISPR-associated proteins), which extensively researched, but recognition of PAM sequences remains restricted. Argonaute, programmable target-activated nuclease, fabricating novel methods due to its unparalleled biological features. In this comprehensive review, we initially elaborate on current nucleic acid testing nucleases, followed by delving into structure nuclease activity system. advantages compared with are highlighted discussed. Furthermore, summarize applications Argonaute-based provide an in-depth analysis future perspectives challenges. Recent research demonstrated that innovative rapidly advancing technology can overcome limitations existing potentially replace them. summary, implementation integration other technologies hold promise developing customized intelligent across various aspects.

Language: Английский

Citations

5
Exploring T7 RNA polymerase-assisted CRISPR/Cas13a amplification for the detection of BNP via electrochemiluminescence sensing platform DOI

Zaiyong Zhang,

Jinglong Li,

Chunlin Chen

et al.

Analytica Chimica Acta, Journal Year: 2024, Volume and Issue: 1300, P. 342409 - 342409

Published: Feb. 26, 2024

Language: Английский

Citations

4
A one-pot CRISPR-RCA strategy for ultrasensitive and specific detection of circRNA DOI

Xinxin Ke,

Ajuan Liang,

Chuanxia Chen

et al.

Analytical Methods, Journal Year: 2024, Volume and Issue: 16(20), P. 3256 - 3262

Published: Jan. 1, 2024

Accurate and precise detection of circular RNA (circRNA) is imperative for its clinical use. However, the inherent challenges in circRNA detection, arising from low abundance potential interference linear isomers, necessitate innovative solutions. In this study, we introduce, first time, application CRISPR/Cas12a system to establish a one-pot, rapid (30 minutes 2 hours), specific ultrasensitive strategy, termed RETA-CRISPR (reverse transcription-rolling circle amplification (RT-RCA) with CRISPR/Cas12a). This method comprises two steps: (1) RT-RCA process amplification, generating repeat units containing back-splicing junction (BSJ) sequences; (2) leveraging protospacer adjacent motif (PAM)-independent Cas12a/crRNA complex precisely recognize target sequences BSJ, thereby initiating collateral cleavage activity Cas12a generate robust fluorescence signal. Remarkably, approach exhibits capability detect circRNAs at concentration as 300 aM. The sensor has been successfully employed accurate hepatocellular carcinoma biomarker hsa_circ_0001445 (circRNA1445) various cell lines. conclusion, seamlessly integrates advantages exponential reaction Cas12a, positioning it compelling tool practical CRISPR-based diagnostics.

Language: Английский

Citations

4