Nucleic acid lateral flow dipstick assay for the duplex detection of and

Abstract Background：Porcine circovirus type 3 (PCV3) is a newly detected pathogen from pigs in the past few years. As the infection rate of PCV3 in the herd is getting higher and wider, the infection is more and more serious. The virus with unclear clinical symptoms and pathogenesis is also paid more and more attention. And detection of pathogenic antibodies has become an important issue. Methods:In this experiment, Loop-mediated isothermal amplification (LAMP) was combined with Lateral flow dipstick (LFD) to establish a rapid and convenient detection method (LAMP-LFD). Results:The results showed that the band was darkened at a nucleic acid concentration of 0.2 pg/ μL using a conventional PCR method, and a clear detection line was observed at a nucleic acid concentration of 0.2 fg/μL using the LAMP-LFD method. The primer and hybridization probe are only compatible with PCV3.Compared to the PCR method, the LAMP-LFD method has a shorter time (about 1 h) and requires less instrumentation. Conclusions: The test results show that the LAMP-LFD method has extremely high sensitivity and specificity. The operation is simple and quick. The visual effect of the detection results is better than that of PCR and single LAMP test, which can bring convenience in practical production applications.

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On-point detection of GM rice in 20 minutes with pullulan as CPA acceleration additive

Analytical Methods ◽

10.1039/c4ay02427c ◽

2014 ◽

Vol 6 (23) ◽

pp. 9198-9201 ◽

Cited By ~ 6

Author(s):

Rui Wang ◽

Jian Wu ◽

Fang Zhang ◽

Liu Wang ◽

Feng Ji

Keyword(s):

Nucleic Acid ◽

Lateral Flow ◽

Isothermal Amplification ◽

Nucleic Acid Amplification ◽

Specific Detection ◽

Lateral Flow Dipstick ◽

Gm Rice ◽

Point Detection

1% (w/v) concentration of pullulan as a cross-priming isothermal amplification additive has an acceleration effect on nucleic acid amplification. With this property, on-point specific detection of DNA by lateral-flow dipstick within 20 minutes was realized.

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Gold-Aptamer-Nanoconstructs Engineered to Detect Conserved Enteroviral Nucleic Acid Sequences

10.26434/chemrxiv.8312324.v1 ◽

2019 ◽

Author(s):

Veeren Chauhan ◽

Mohamed M Elsutohy ◽

C Patrick McClure ◽

Will Irving ◽

Neil Roddis ◽

...

Keyword(s):

Nucleic Acid ◽

In Silico ◽

Point Of Care ◽

Lateral Flow ◽

Nucleic Acid Sequence ◽

In Silico Screening ◽

Lateral Flow Assays ◽

Life Threatening ◽

Software And Hardware ◽

Nucleic Acid Sequences

Enteroviruses are a ubiquitous mammalian pathogen that can produce mild to life-threatening disease. Bearing this in mind, we have developed a rapid, accurate and economical point-of-care biosensor that can detect a nucleic acid sequences conserved amongst 96% of all known enteroviruses. The biosensor harnesses the physicochemical properties of gold nanoparticles and aptamers to provide colourimetric, spectroscopic and lateral flow-based identification of an exclusive enteroviral RNA sequence (23 bases), which was identified through in silico screening. Aptamers were designed to demonstrate specific complementarity towards the target enteroviral RNA to produce aggregated gold-aptamer nanoconstructs. Conserved target enteroviral nucleic acid sequence (≥ 1x10-7 M, ≥1.4×10-14 g/mL), initiates gold-aptamer-nanoconstructs disaggregation and a signal transduction mechanism, producing a colourimetric and spectroscopic blueshift (544 nm (purple) > 524 nm (red)). Furthermore, lateral-flow-assays that utilise gold-aptamer-nanoconstructs were unaffected by contaminating human genomic DNA, demonstrated rapid detection of conserved target enteroviral nucleic acid sequence (< 60 s) and could be interpreted with a bespoke software and hardware electronic interface. We anticipate our methodology will translate in-silico screening of nucleic acid databases to a tangible enteroviral desktop detector, which could be readily translated to related organisms. This will pave-the-way forward in the clinical evaluation of disease and complement existing strategies at overcoming antimicrobial resistance.

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Recombinase-aided amplification–lateral flow dipstick assay—a specific and sensitive method for visual detection of avian infectious laryngotracheitis virus

Poultry Science ◽

10.1016/j.psj.2020.12.008 ◽

2020 ◽

Author(s):

Wenjing Wang ◽

Chunguang Wang ◽

Zichuang Zhang ◽

Peng Zhang ◽

Xianghe Zhai ◽

...

Keyword(s):

Visual Detection ◽

Lateral Flow ◽

Infectious Laryngotracheitis Virus ◽

Lateral Flow Dipstick ◽

Infectious Laryngotracheitis ◽

Laryngotracheitis Virus ◽

Sensitive Method

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Recombinase-Aided Amplification Coupled with Lateral Flow Dipstick for Efficient and Accurate Detection of Porcine Parvovirus

Life ◽

10.3390/life11080762 ◽

2021 ◽

Vol 11 (8) ◽

pp. 762

Author(s):

Yihong He ◽

Wenxian Chen ◽

Jindai Fan ◽

Shuangqi Fan ◽

Hongxing Ding ◽

...

Keyword(s):

Detection Method ◽

Cross Reactivity ◽

Lateral Flow ◽

Porcine Parvovirus ◽

Swine Industry ◽

Resource Limited ◽

Efficient Detection ◽

Lateral Flow Dipstick ◽

Polymerase Chain ◽

Embryonic Death

Porcine parvovirus (PPV) infection is the primary cause of SMEDI (stillbirth; mummification; embryonic death; infertility) syndrome, which is a global burden for the swine industry. Thus, it is crucial to establish a rapid and efficient detection method against PPV infection. In the present work, we developed a recombinase-aided amplification (RAA) assay, coupled with a lateral flow dipstick (LFD), to achieve an amplification of PPV DNA at 37 °C within 15 min. The detection limits of PPV RAA-LFD assay were 102 copies/μL recombinant plasmid pMD19-T-VP1, 6.38 × 10−7 ng/μL PPV DNA, and 10−1 TCID50/mL virus, respectively. This method was highly specific for PPV detection with no cross-reactivity for other swine pathogens. In contrast to polymerase chain reaction (PCR), the PPV RAA-LFD assay is more sensitive and cost-saving. Hence, the established PPV RAA-LFD assay provided an alternative for PPV detection, especially in resource-limited regions.

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