Dataset Information

Simultaneous Rapid Nucleic Acid and Protein Detection in a Lateral Chromatography Chip for COVID-19 Diagnosis.

ABSTRACT: In this work, we report a fast, portable, and economical microfluidic platform for the simultaneous detection of nucleic acid and proteins. Using SARS-CoV-2 as a target, this microfluidic chip enabled to simultaneously detect the SARS-CoV-2 RNA (N gene) antigen (or specific IgG antibody) with respective detection limits of 1 copy/μL for nucleic acid, 0.85 ng/mL for antigen, and 5.80 ng/mL for IgG within 30 min with high stability and anti-interference ability. The capability of this system in clinical applications was further evaluated using clinical samples, displaying 100% sensitivity and 100% specificity for COVID-19 diagnosis. These findings demonstrate the potential of this method to be used for the detection and subsequent control of pathogens.

SUBMITTER: Lin Q

PROVIDER: S-EPMC9631717 | biostudies-literature | 2022 Nov

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Simultaneous Rapid Nucleic Acid and Protein Detection in a Lateral Chromatography Chip for COVID-19 Diagnosis.

Lin Qiuyuan Q Zhang Jin J Liu Liling L Kong Jilie J Fang Xueen X

ACS omega 20221021 43

In this work, we report a fast, portable, and economical microfluidic platform for the simultaneous detection of nucleic acid and proteins. Using SARS-CoV-2 as a target, this microfluidic chip enabled to simultaneously detect the SARS-CoV-2 RNA (N gene) antigen (or specific IgG antibody) with respective detection limits of 1 copy/μL for nucleic acid, 0.85 ng/mL for antigen, and 5.80 ng/mL for IgG within 30 min with high stability and anti-interference ability. The capability of this system in cl ...[more]

PMID: 36340144

Similar Datasets

Project description:The increasing prevalence of antimicrobial resistance is a serious threat to global public health. One of the most concerning trends is the rapid spread of Carbapenemase-Producing Organisms (CPO), where colistin has become the last-resort antibiotic treatment. The emergence of colistin resistance, including the spread of mobilized colistin resistance (mcr) genes, raises the possibility of untreatable bacterial infections and motivates the development of improved diagnostics for the detection of colistin-resistant organisms. This work demonstrates a rapid response for detecting the most recently reported mcr gene, mcr-9, using a portable and affordable lab-on-a-chip (LoC) platform, offering a promising alternative to conventional laboratory-based instruments such as real-time PCR (qPCR). The platform combines semiconductor technology, for non-optical real-time DNA sensing, with a smartphone application for data acquisition, visualization and cloud connectivity. This technology is enabled by using loop-mediated isothermal amplification (LAMP) as the chemistry for targeted DNA detection, by virtue of its high sensitivity, specificity, yield, and manageable temperature requirements. Here, we have developed the first LAMP assay for mcr-9 - showing high sensitivity (down to 100 genomic copies/reaction) and high specificity (no cross-reactivity with other mcr variants). This assay is demonstrated through supporting a hospital investigation where we analyzed nucleic acids extracted from 128 carbapenemase-producing bacteria isolated from clinical and screening samples and found that 41 carried mcr-9 (validated using whole genome sequencing). Average positive detection times were 6.58 ± 0.42 min when performing the experiments on a conventional qPCR instrument (n = 41). For validating the translation of the LAMP assay onto a LoC platform, a subset of the samples were tested (n = 20), showing average detection times of 6.83 ± 0.92 min for positive isolates (n = 14). All experiments detected mcr-9 in under 10 min, and both platforms showed no statistically significant difference (p-value > 0.05). When sample preparation and throughput capabilities are integrated within this LoC platform, the adoption of this technology for the rapid detection and surveillance of antimicrobial resistance genes will decrease the turnaround time for DNA detection and resistotyping, improving diagnostic capabilities, patient outcomes, and the management of infectious diseases.

Project description:MicroRNA122 (miR-122) is a microRNA that is highly expressed in hepatocytes and has been identified as a prospective therapeutic target and biomarker for liver injury. An expanding body of research has demonstrated that miR-122 is a critical regulator in both the initiation and progression of a wide range of liver diseases. Traditional methods for detecting miR-122 mainly include Northern blotting and qRT-PCR, but they are technically complex and cumbersome, requiring expensive instruments and high technical requirements. In this paper, we present a novel rapid testing method utilizing a lateral flow nucleic acid biosensor (LFNAB) for the sensitive and time-efficient detection of miR-122. This approach offers several advantages, including a high specificity for miR-122, the ability to detect low concentrations of the target molecule, and a significantly reduced testing time compared to conventional detection methods. In this study, a thiol-modified single-stranded detection DNA probe (Det-DNA), a biotinylated single-stranded capture DNA probe (Cap-DNA), and a biotinylated single-stranded control DNA probe (Con-DNA) are used to construct the LFNAB. A gold nanoparticle (AuNP) is a colored tag, which is used to label the Det-DNA probe. The principle of detecting miR-122 is based on dual DNA-miRNA hybridization reactions on the LFNAB to form sandwich-type AuNP-Det-DNA-miR-122-Cap-DNA complexes, which are captured on the test area of LFNAB for visualization and quantification. After systematic optimization of conditions of experiment, the response of LFNAB was highly linear within the scope of 0 pM-100 pM miR-122, and the detection limit in 15 min was 3.90 pM. The use of LFNAB to detect miR-122 in serum and fingertip blood has yielded satisfactory results. This successful application indicates the effectiveness of LFNAB in detecting miR-122 in both serum and fingertip blood samples, showcasing its potential utility in clinical and research settings for assessing miR-122 levels in different biological samples.

Project description:Global pandemics such as COVID-19 have resulted in significant global social and economic disruption. Although polymerase chain reaction (PCR) is recommended as the standard test for identifying the SARS-CoV-2, conventional assays are time-consuming. In parallel, although artificial intelligence (AI) has been employed to contain the disease, the implementation of AI in PCR analytics, which may enhance the cognition of diagnostics, is quite rare. The information that the amplification curve reveals can reflect the dynamics of reactions. Here, we present a novel AI-aided on-chip approach by integrating deep learning with microfluidic paper-based analytical devices (µPADs) to detect synthetic RNA templates of the SARS-CoV-2 ORF1ab gene. The µPADs feature a multilayer structure by which the devices are compatible with conventional PCR instruments. During analysis, real-time PCR data were synchronously fed to three unsupervised learning models with deep neural networks, including RNN, LSTM, and GRU. Of these, the GRU is found to be most effective and accurate. Based on the experimentally obtained datasets, qualitative forecasting can be made as early as 13 cycles, which significantly enhances the efficiency of the PCR tests by 67.5% (∼40 min). Also, an accurate prediction of the end-point value of PCR curves can be obtained by GRU around 20 cycles. To further improve PCR testing efficiency, we also propose AI-aided dynamic evaluation criteria for determining critical cycle numbers, which enables real-time quantitative analysis of PCR tests. The presented approach is the first to integrate AI for on-chip PCR data analysis. It is capable of forecasting the final output and the trend of qPCR in addition to the conventional end-point Cq calculation. It is also capable of fully exploring the dynamics and intrinsic features of each reaction. This work leverages methodologies from diverse disciplines to provide perspectives and insights beyond the scope of a single scientific field. It is universally applicable and can be extended to multiple areas of fundamental research. Graphical abstract Image, graphical abstract

Dataset Information

Simultaneous Rapid Nucleic Acid and Protein Detection in a Lateral Chromatography Chip for COVID-19 Diagnosis.

Publications

Simultaneous Rapid Nucleic Acid and Protein Detection in a Lateral Chromatography Chip for COVID-19 Diagnosis.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets