Dataset Information

Direct Detection of Unamplified Pathogen RNA in Blood Lysate using an Integrated Lab-in-a-Stick Device and Ultrabright SERS Nanorattles.

ABSTRACT: Direct detection of genetic biomarkers in body fluid lysate without target amplification will revolutionize nucleic acid-based diagnostics. However, the low concentration of target sequences makes this goal challenging. We report a method for direct detection of pathogen RNA in blood lysate using a bioassay using surface-enhanced Raman spectroscopy (SERS)-based detection integrated in a "lab-in-a-stick" portable device. Two levels of signal enhancement were employed to achieve the sensitivity required for direct detection. Each target sequence was tagged with an ultrabright SERS-encoded nanorattle with ultrahigh SERS signals, and these tagged target sequences were concentrated into a focused spot for detection using hybridization sandwiches with magnetic microbeads. Furthermore, the washing process was automated by integration into a "lab-in-a-stick" portable device. We could directly detect synthetic target with a limit of detection of 200?fM. More importantly, we detected plasmodium falciparum malaria parasite RNA directly in infected red blood cells lysate. To our knowledge, this is the first report of SERS-based direct detection of pathogen nucleic acid in blood lysate without nucleic acid extraction or target amplification. The results show the potential of our integrated bioassay for field use and point-of-care diagnostics.

SUBMITTER: Ngo HT

PROVIDER: S-EPMC5840326 | biostudies-literature | 2018 Mar

REPOSITORIES: biostudies-literature

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Direct Detection of Unamplified Pathogen RNA in Blood Lysate using an Integrated Lab-in-a-Stick Device and Ultrabright SERS Nanorattles.

Ngo Hoan T HT Freedman Elizabeth E Odion Ren Abelard RA Strobbia Pietro P De Silva Indrasekara Agampodi Swarnapali AS Vohra Priya P Taylor Steve M SM Vo-Dinh Tuan T

Scientific reports 20180306 1

Direct detection of genetic biomarkers in body fluid lysate without target amplification will revolutionize nucleic acid-based diagnostics. However, the low concentration of target sequences makes this goal challenging. We report a method for direct detection of pathogen RNA in blood lysate using a bioassay using surface-enhanced Raman spectroscopy (SERS)-based detection integrated in a "lab-in-a-stick" portable device. Two levels of signal enhancement were employed to achieve the sensitivity re ...[more]

PMID: 29511216

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Project description:Digital droplet PCR (ddPCR) is a powerful amplification technique for absolute quantification of viral nucleic acids. Although commercial ddPCR devices are effective in the lab bench tests, they cannot meet current urgent requirements for on-site and rapid screening for patients. Here, we have developed a portable and fully integrated lab-on-a-disc (LOAD) device for quantitively screening infectious disease agents. Our designed LOAD device has integrated (i) microfluidics chips, (ii) a transparent circulating oil-based heat exchanger, and (iii) an on-disc transmitted-light fluorescent imaging system into one compact and portable box. Thus, droplet generation, PCR thermocycling, and analysis can be achieved in a single LOAD device. This feature is a significant attribute for the current clinical application of disease screening. For this custom-built ddPCR setup, we have first demonstrated the loading and ddPCR amplification ability by using influenza A virus-specific DNA fragments with different concentrations (diluted from the original concentration to 107 times), followed by analyzing the droplets with an external fluorescence microscope as a standard calibration test. The measured DNA concentration is linearly related to the gradient-dilution factor, which validated the precise quantification for the samples. In addition to the calibration tests using DNA fragments, we also employed this ddPCR-LOAD device for clinical samples with different viruses. Infectious samples containing five different viruses, including influenza A virus (IAV), respiratory syncytial virus (RSV), varicella zoster virus (VZV), Zika virus (ZIKV), and adenovirus (ADV), were injected into the device, followed by analyzing the droplets with an external fluorescence microscope with the lowest detected concentration of 20.24 copies/µL. Finally, we demonstrated the proof-of-concept detection of clinical samples of IAV using the on-disc fluorescence imaging system in our fully integrated device, which proves the capability of this device in clinical sample detection. We anticipate that this integrated ddPCR-LOAD device will become a flexible tool for on-site disease detection.

Project description:BackgroundWhile e-cigarettes have been identified as an effective means of tobacco harm reduction, the degree to which these devices will realise their harm reduction potential will be determined in large part by how available these products are to adults who smoke. One of the key factors determining that availability is the regulatory regime around these products. Within the US e-cigarettes have become the most commonly used tobacco product by middle and high school students, with disposable e-cigarettes now the most popular type of device used by youth. In this paper, we report data on the prevalence with which one of the most popular disposable e-cigarettes within the US is being used by youth (aged 13-17) and underage young adults (aged 18-20).MethodsA cross-sectional online survey of a probability-based sample of 1215 youth (13-17 years) recruited via Ipsos' KnowledgePanel and 3370 young adults aged 18-24, among whom 1125 were aged 18-20, recruited via Ipsos' KnowledgePanel and online consumer research panels.ResultsAmong youth, 3.50% (95% CI: 2.46-4.83) reported smoking combustible cigarettes in the past 30 days, and 6.73% (95% CI: 5.26-8.48) reported using an e-cigarette in the past 30 days. Among underage young adults, 7.22% (95% CI: 5.24-9.67) reported smoking combustible cigarettes every day or some days, and 15.90% (95% CI: 12.96-19.21) reported using e-cigarettes every day or some days. Despite the scale of e-cigarette use in general among the US youth, only 0.04% (95% CI: 0.00-0.38) of 13-17 years old reported using the BIDI® Stick disposable e-cigarette in the past 30 days.ConclusionsWhile disposable e-cigarettes have become the most popular type of e-cigarette used by the US youth, it is likely that the prevalence of use of individual devices varies significantly. There is a need to regularly monitor the use of e-cigarettes by type of device and brand, in order to determine which products have the greatest potential to reduce smoking-related harm among adults who smoke and which products are being used by youth and underage young adults.

Dataset Information

Direct Detection of Unamplified Pathogen RNA in Blood Lysate using an Integrated Lab-in-a-Stick Device and Ultrabright SERS Nanorattles.

Publications

Direct Detection of Unamplified Pathogen RNA in Blood Lysate using an Integrated Lab-in-a-Stick Device and Ultrabright SERS Nanorattles.

Similar Datasets

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